U.S. patent application number 11/503245 was filed with the patent office on 2007-03-29 for novel peptide compositions and their use in particular in the preparation of pharmaceutical compositions active against the hepatitis c virus.
Invention is credited to Anne Fournillier, Nourredine Himoudi, Genevieve Inchauspe, Perrine Martin.
Application Number | 20070072176 11/503245 |
Document ID | / |
Family ID | 29286584 |
Filed Date | 2007-03-29 |
United States Patent
Application |
20070072176 |
Kind Code |
A1 |
Inchauspe; Genevieve ; et
al. |
March 29, 2007 |
Novel peptide compositions and their use in particular in the
preparation of pharmaceutical compositions active against the
hepatitis C virus
Abstract
The invention relates to peptide compositions for use in the
preparation of pharmaceutical compositions against hepatitis C
virus, as well as corresponding methods.
Inventors: |
Inchauspe; Genevieve; (Lyon,
FR) ; Fournillier; Anne; (Lyon, FR) ; Himoudi;
Nourredine; (Lyon, FR) ; Martin; Perrine;
(Decines Charpieu, FR) |
Correspondence
Address: |
CLARK & ELBING LLP
101 FEDERAL STREET
BOSTON
MA
02110
US
|
Family ID: |
29286584 |
Appl. No.: |
11/503245 |
Filed: |
August 11, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10514762 |
Nov 16, 2004 |
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PCT/FR03/01478 |
May 15, 2003 |
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11503245 |
Aug 11, 2006 |
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Current U.S.
Class: |
435/5 ;
424/189.1; 435/325; 435/456; 435/6.16; 435/69.3; 530/350;
530/388.3; 536/23.72 |
Current CPC
Class: |
A61K 39/00 20130101;
C12N 2770/24222 20130101; C07K 14/005 20130101; A61P 31/00
20180101; A61P 31/14 20180101; A61P 37/04 20180101; A61K 38/00
20130101 |
Class at
Publication: |
435/005 ;
435/006; 435/069.3; 435/456; 435/325; 530/350; 536/023.72;
530/388.3; 424/189.1 |
International
Class: |
C12Q 1/70 20060101
C12Q001/70; C12Q 1/68 20060101 C12Q001/68; C07H 21/04 20060101
C07H021/04; A61K 39/29 20060101 A61K039/29; C07K 14/18 20060101
C07K014/18; C07K 16/10 20060101 C07K016/10 |
Foreign Application Data
Date |
Code |
Application Number |
May 17, 2002 |
FR |
02/06111 |
Claims
1-88. (canceled)
89. Fusion peptide characterized in that it comprises a fusion of
at least two peptides chosen from the group consisting of A, B, C,
and D peptides, B' and C' epitopes, said A peptide having at least
the following amino acid sequence SEQ ID NO: 1:
X.sub.1AX.sub.2QGYKVX.sub.3VLNPSVX.sub.4ATLX.sub.5FGX.sub.6YMSKAX.sub.7GX-
.sub.8, in which X.sub.1 is Y or H, X.sub.2 is A, G or T, X.sub.3
is R or L, X.sub.4 is A or T, X.sub.5 is G or S, X.sub.6 is A, T or
V, X.sub.7 is H or Y and X.sub.8 is I, T, M, or V, and at most
sequence the following amino acid sequence SEQ ID NO: 2,
SX.sub.9X.sub.10VPX.sub.11X.sub.12X.sub.1AX.sub.2QGYKVX.sub.3VLNPSVX.sub.-
4ATLX.sub.5FGX.sub.6YMSKAX.sub.7GX.sub.8X.sub.13P
X.sub.14X.sub.15X.sub.16X.sub.17GV, in which X.sub.1 to X.sub.8 are
as defined above, and X.sub.9 is T or N, X.sub.10 is K or R,
X.sub.11 is A or V, X.sub.12 is A or E, X.sub.13 is E or D,
X.sub.14 is N or S, X.sub.15 is I, L or V, X.sub.16 is R or S, and
X.sub.17 is T or S, said B peptide having at least the following
amino acid sequence SEQ ID NO: 45:
GX.sub.18X.sub.19X.sub.20X.sub.21X.sub.22X.sub.23TSLTGRDX.sub.24NX.s-
ub.25X.sub.26X.sub.27GEX.sub.28QX.sub.29X.sub.30STAX.sub.31X.sub.32X.sub.3-
3FL X.sub.34X.sub.35X.sub.36X.sub.37NGX.sub.38X.sub.39WTVX.sub.40
in which X.sub.18 is L or V, X.sub.19 is L or F, X.sub.20 is G or
S, X.sub.21 is C or T, X.sub.22 is I or V, X.sub.23 is I or V,
X.sub.24 is K, R or T, X.sub.25 is Q or E, X.sub.26 is V or N,
X.sub.27 is D, E or C, X.sub.28 is V or A, X.sub.29 is V, I, E or
M, X.sub.30 is L or V, X.sub.31 is T, K or A, X.sub.32 is Q or H,
X.sub.33 is S or T, X.sub.34 is A or G, X.sub.35 is T or S,
X.sub.36 is C or A, X.sub.37 is V, I or T, X.sub.38 is V or A,
X.sub.39 is C or M, and X.sub.40 is Y or F, and at most the
following sequence SEQ ID NO: 46:
AX.sub.41TX.sub.42YX.sub.43X.sub.44QTRGX.sub.18X.sub.19X.sub.20X.sub.21X.-
sub.22X.sub.23TSLTGRDX.sub.24NX.sub.25X.sub.26X.sub.27GEX.sub.28Q
X.sub.29X.sub.30STAX.sub.31X.sub.32X.sub.33FLX.sub.34X.sub.3X.sub.36X.sub-
.37NGX.sub.38X.sub.39WTVX.sub.40 HGAGX.sub.45X.sub.4X.sub.7 in
which X.sub.18 to X.sub.40 are as defined above and X.sub.41 is P,
S or H, X.sub.42 is A or T, X.sub.43 is S, A, T or C, X.sub.44 is Q
or R, X.sub.45 is S, T or A, X.sub.46 is K or R, and X.sub.47 is T
or I, said C peptide having at least the following amino acid
sequence SEQ ID NO: 127: SX.sub.47'M
X.sub.48FTX.sub.49X.sub.50X.sub.51
TSPLX.sub.52X.sub.53X.sub.54X.sub.55TLX.sub.56FNIX.sub.57GGWVAX.sub.58QX.-
sub.59 in which X.sub.47' is L or P, X.sub.48 is A or S, X.sub.49
is A or S, X.sub.50 is A or S, X.sub.51 is I or V, X.sub.52 is T, S
or A, X.sub.53 is T or I, X.sub.54 is Q, S or G, X.sub.55 is N, Q,
H, S, Y or T, X.sub.56 is L or M, X.sub.57 is L or W, X.sub.58 is A
or S, and X.sub.59 is L, P or I, and at most the following sequence
SEQ ID NO: 128: NFIX.sub.60GX.sub.61
QYLAX.sub.62LSTLPGNX.sub.63AX.sub.64X.sub.65SX.sub.47'MX.sub.48FTX.sub.49-
X.sub.50X.sub.51TSPLX.sub.52X.sub.53X.sub.54X.sub.55TLX.sub.56FNIX.sub.57G-
GWVAX.sub.58QX.sub.59X.sub.66X.sub.67X.sub.68 in which X.sub.47' to
X.sub.59 are as defined above and X.sub.60 is T or S, X.sub.61 is
I, T or V, X.sub.62 is G or A, X.sub.63 is P or L, X.sub.64 is I or
M, X.sub.65 is A, V or R, X.sub.66 is A or R, X.sub.67 is P, A, or
D, and X.sub.68 is P, A, or S, said D peptide having at least the
following amino acid sequence SEQ ID NO: 174:
X.sub.69KX.sub.70ARX.sub.71IVX.sub.72PX.sub.73LGX.sub.74RVCEKX.sub.75ALX.-
sub.76X.sub.77VX.sub.78X.sub.79X.sub.80X.sub.81 in which X.sub.69
is R or Q, X.sub.70 is P or A, X.sub.71 is L or F, X.sub.72 is F or
Y, X.sub.73 is D or E, X.sub.74 is V or S, X.sub.75 is M or R,
X.sub.76 is Y or H, X.sub.77 is D or N, X.sub.78 is V or I,
X.sub.79 is S, T or K, X.sub.80 is T, K, I or N and X.sub.81 is L
or T, and at most the following amino acid sequence SEQ ID NO: 175:
KGGX.sub.69KX.sub.70ARX.sub.711VX.sub.72PX.sub.73LGX.sub.74RVCEKX.sub.75A-
LX.sub.76X.sub.77VX.sub.78X.sub.79X.sub.80X.sub.81X.sub.82X.sub.83X.sub.84-
VMG X.sub.85X.sub.86YX.sub.87X.sub.88Q in which X.sub.69 to
X.sub.8, are as defined above and X.sub.82 is P or A, X.sub.83 is
Q, L, H, R, K or P, X.sub.84 is A, T, V or P, X.sub.85 is P, S or
A, X.sub.86 is S or A, X.sub.87 is G or R and X.sub.88 is F or C,
said B' epitope having the following sequence SEQ ID NO: 213:
GX.sub.18X.sub.19X.sub.20X.sub.21X.sub.22X.sub.23TSL in which
X.sub.18 is L or V, X.sub.19 is L or F, X.sub.20 is G or S,
X.sub.21 is C or T, X.sub.22 is I or V and X.sub.23 is I or V, and
said C' epitope having the following sequence SEQ ID NO: 221:
SPLX.sub.52X.sub.53X.sub.54X.sub.55TL in which X.sub.52 is T, S or
A, X.sub.53 is T or I, X.sub.54 is Q, S or G and X.sub.55 is N, Q,
H, S, Y or T.
90. The fusion peptide according to claim 89, wherein the A peptide
is chosen from: the peptides having at least sequence SEQ ID NO: 3
and at most sequence SEQ ID NO: 19, the peptides having at least
sequence SEQ ID NO: 4 and at most a sequence chosen from sequences
SEQ ID NO: 20, SEQ ID NO: 24, SEQ ID NO: 32 and SEQ ID NO: 34, the
peptides having at least sequence SEQ ID NO: 5 and at most a
sequence chosen from sequences SEQ ID NO: 21, SEQ ID NO: 28 and SEQ
ID NO: 36, the peptides having at least sequence SEQ ID NO: 6 and
at most a sequence chosen from sequences SEQ ID NO: 22, SEQ ID NO:
27 and SEQ ID NO: 41, the peptides having at least sequence SEQ ID
NO: 7 and at most a sequence chosen from sequences SEQ ID NO: 23
and SEQ ID NO: 37, the peptides having at least sequence SEQ ID NO:
8 and at most sequence SEQ ID NO: 25, the peptides having at least
sequence SEQ ID NO: 9 and at most sequence SEQ ID NO: 26, the
peptides having at least sequence SEQ ID NO: 10 and at most
sequence SEQ ID NO: 29, the peptides having at least sequence SEQ
ID NO: 11 and at most sequence SEQ ID NO: 30, the peptides having
at least sequence SEQ ID NO: 12 and at most sequence SEQ ID NO: 31,
the peptides having at least sequence SEQ ID NO: 13 and at most
sequence SEQ ID NO: 33, the peptides having at least sequence SEQ
ID NO: 14 and at most a sequence chosen from sequences SEQ ID NO:
35 and SEQ ID NO: 39, the peptides having at least sequence SEQ ID
NO: 15 and at most sequence SEQ ID NO: 38, the peptides having at
least sequence SEQ ID NO: 16 and at most a sequence chosen from
sequences SEQ ID NO: 40 and SEQ ID NO: 42, the peptides having at
least sequence SEQ ID NO: 17 and at most sequence SEQ ID NO: 43,
and the peptides having at least sequence SEQ ID NO: 18 and at most
sequence SEQ ID NO: 44.
91. The fusion peptide according to claim 89, wherein the A peptide
is chosen from the peptides of sequences SEQ ID NO: 3 to 18.
92. The fusion peptide according to claim 89, wherein the B peptide
is chosen from: the peptides having at least sequence SEQ ID NO: 47
and at most sequence SEQ ID NO: 81, the peptides having at least
sequence SEQ ID NO: 48 and at most sequence SEQ ID NO: 82, the
peptides having at least sequence SEQ ID NO: 49 and at most
sequence SEQ ID NO: 83, the peptides having at least sequence SEQ
ID NO: 50 and at most a sequence chosen from sequences SEQ ID NO:
84, SEQ ID NO: 90, SEQ ID NO: 105, SEQ ID NO: 107 and SEQ ID NO:
116, the peptides having at least sequence SEQ ID NO: 51 and at
most a sequence chosen from sequences SEQ ID NO: 85 and SEQ ID NO:
124, the peptides having at least sequence SEQ ID NO: 52 and at
most a sequence chosen from sequences SEQ ID NO: 86 and SEQ ID NO:
120, the peptides having at least sequence SEQ ID NO: 53 and at
most sequence SEQ ID NO: 87, the peptides having at least sequence
SEQ ID NO: 54 and at most a sequence chosen from sequences SEQ ID
NO: 88 and SEQ ID NO: 117, the peptides having at least sequence
SEQ ID NO: 55 and at most sequence SEQ ID NO: 89, the peptides
having at least sequence SEQ ID NO: 56 and at most a sequence
chosen from sequences SEQ ID NO: 91 and SEQ ID NO: 92, the peptides
having at least sequence SEQ ID NO: 57 and at most sequence SEQ ID
NO: 93, the peptides having at least sequence SEQ ID NO: 58 and at
most a sequence chosen from sequences SEQ ID NO: 94 and SEQ ID NO:
110, the peptides having at least sequence SEQ ID NO: 59 and at
most sequence SEQ ID NO: 95, the peptides having at least sequence
SEQ ID NO: 60 and at most a sequence chosen from sequences SEQ ID
NO: 96 and SEQ ID NO: 115, the peptides having at least sequence
SEQ ID NO: 61 and at most sequence SEQ ID NO: 97, the peptides
having at least sequence SEQ ID NO: 62 and at most a sequence
chosen from sequences SEQ ID NO: 98 and SEQ ID NO: 109, the
peptides having at least sequence SEQ ID NO: 63 and at most
sequence SEQ ID NO: 99, the peptides having at least sequence SEQ
ID NO: 64 and at most sequence SEQ ID NO: 100, the peptides having
at least sequence SEQ ID NO: 65 and at most sequence SEQ ID NO:
101, the peptides having at least sequence SEQ ID NO: 66 and at
most sequence SEQ ID NO: 102, the peptides having at least sequence
SEQ ID NO: 67 and at most sequence SEQ ID NO: 103, the peptides
having at least sequence SEQ ID NO: 68 and at most sequence SEQ ID
NO: 104, the peptides having at least sequence SEQ ID NO: 69 and at
most sequence SEQ ID NO: 106, the peptides having at least sequence
SEQ ID NO: 70 and at most sequence SEQ ID NO: 108, the peptides
having at least sequence SEQ ID NO: 71 and at most sequence SEQ ID
NO: 111, the peptides having at least sequence SEQ ID NO: 72 and at
most sequence SEQ ID NO: 112, the peptides having at least sequence
SEQ ID NO: 73 and at most sequence SEQ ID NO: 113, the peptides
having at least sequence SEQ ID NO: 74 and at most sequence SEQ ID
NO: 114, the peptides having at least sequence SEQ ID NO: 75 and at
most a sequence chosen from sequences SEQ ID NO: 118 and SEQ ID NO:
119, the peptides having at least sequence SEQ ID NO: 76 and at
most sequence SEQ ID NO: 121, the peptides having at least sequence
SEQ ID NO: 77 and at most sequence SEQ ID NO: 122, the peptides
having at least sequence SEQ ID NO: 78 and at most sequence SEQ ID
NO: 123, the peptides having at least sequence SEQ ID NO: 79 and at
most sequence SEQ ID NO: 125, and the peptides having at least
sequence SEQ ID NO: 80 and at most sequence SEQ ID NO: 126.
93. The fusion peptide according to claim 89, wherein the B peptide
is chosen from the peptides of sequences SEQ ID NO: 47 to 80.
94. The fusion peptide according to claim 89, wherein the C peptide
is chosen from: the peptides having at least sequence SEQ ID NO:
129 and at most a sequence chosen from sequences SEQ ID NO: 147,
SEQ ID NO: 153, SEQ ID NO: 162 and SEQ ID NO: 167, the peptides
having at least sequence SEQ ID NO: 130 and at most a sequence
chosen from sequences SEQ ID NO: 148 and SEQ ID NO: 150, the
peptides having at least sequence SEQ ID NO: 131 and at most
sequence SEQ ID NO: 149, the peptides having at least sequence SEQ
ID NO: 132 and at most a sequence chosen from sequences SEQ ID NO:
151, SEQ ID NO: 155, SEQ ID NO: 168 and SEQ ID NO: 171, the
peptides having at least sequence SEQ ID NO: 133 and at most
sequence SEQ ID NO: 152, the peptides having at least sequence SEQ
ID NO: 134 and at most a sequence chosen from sequences SEQ ID NO:
154 and SEQ ID NO: 169, the peptides having at least sequence SEQ
ID NO: 135 and at most sequence SEQ ID NO: 156, the peptides having
at least sequence SEQ ID NO: 136 and at most a sequence chosen from
sequences SEQ ID NO: 157 and SEQ ID NO: 170, the peptides having at
least sequence SEQ ID NO: 137 and at most sequence SEQ ID NO: 158,
the peptides having at least sequence SEQ ID NO: 138 and at most
sequence SEQ ID NO: 159, the peptides having at least sequence SEQ
ID NO: 139 and at most sequence SEQ ID NO: 160, the peptides having
at least sequence SEQ ID NO: 140 and at most sequence SEQ ID NO:
161, the peptides having at least sequence SEQ ID NO: 141 and at
most sequence SEQ ID NO: 163, the peptides having at least sequence
SEQ ID NO: 142 and at most sequence SEQ ID NO: 164, the peptides
having at least sequence SEQ ID NO: 143 and at most sequence SEQ ID
NO: 165, the peptides having at least sequence SEQ ID NO: 144 and
at most sequence SEQ ID NO: 166, the peptides having at least
sequence SEQ ID NO: 145 and at most sequence SEQ ID NO: 172, and
the peptides having at least sequence SEQ ID NO: 146 and at most
sequence SEQ ID NO: 173.
95. The fusion peptide according to claim 89, wherein the C peptide
is chosen from the peptides of sequences SEQ ID NO: 129 to 146.
96. The fusion peptide according to claim 89, wherein the D peptide
is chosen from: the peptides having at least sequence SEQ ID NO:
176 and at most a sequence chosen from sequences SEQ ID NO: 188,
SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID NO: 194, SEQ ID NO: 201, SEQ
ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ ID NO: 205 and SEQ
ID NO: 210, the peptides having at least sequence SEQ ID NO: 177
and at most a sequence chosen from sequences SEQ ID NO: 189 and SEQ
ID NO: 190, the peptides having at least sequence SEQ ID NO: 178
and at most sequence SEQ ID NO: 191, the peptides having at least
sequence SEQ ID NO: 179 and at most sequence SEQ ID NO: 195, the
peptides having at least sequence SEQ ID NO: 180 and at most a
sequence chosen from sequences SEQ ID NO: 196 and SEQ ID NO: 206,
the peptides having at least sequence SEQ ID NO: 181 and at most
sequence SEQ ID NO: 197, the peptides having at least sequence SEQ
ID NO: 182 and at most a sequence chosen from sequences SEQ ID NO:
198 and SEQ ID NO: 209, the peptides having at least sequence SEQ
ID NO: 183 and at most a sequence chosen from sequences SEQ ID NO:
199 and SEQ ID NO: 200, the peptides having at least sequence SEQ
ID NO: 184 and at most sequence SEQ ID NO: 207, the peptides having
at least sequence SEQ ID NO: 185 and at most sequence SEQ ID NO:
208, the peptides having at least sequence SEQ ID NO: 186 and at
most sequence SEQ ID NO: 211, and the peptides having at least
sequence SEQ ID NO: 187 and at most sequence SEQ ID NO: 212.
97. The fusion peptide according to claim 89, wherein the D peptide
is chosen from the peptides of sequences SEQ ID NO: 176 to 187.
98. The fusion peptide according to claim 89, wherein the B'
epitope is chosen from the epitopes of sequences SEQ ID NO: 214 to
SEQ ID NO: 220.
99. The fusion peptide according to claim 89, wherein the C'
epitope is chosen from the epitopes of sequences SEQ ID NO: 222 to
SEQ ID NO: 232.
100. The fusion peptide according to claim 89, characterized in
that it comprises the fusion of two peptides chosen among: i) the A
peptide as defined in claim 89 and another peptide chosen from the
B to D peptides as defined in claim 89, or ii) the B peptide as
defined in claim 89 and another peptide chosen from the A, C or D
peptides as defined in claim 89, or iii) the C peptide as defined
in claim 89 and another peptide chosen from the A, B or D peptides
as defined in claim 89, or iv) the D peptide as defined in claim 89
and another peptide chosen from the A, B or C peptides as defined
in claim 89.
101. The fusion peptide according to claim 89, characterized in
that it comprises the fusion of three peptides chosen among: i) the
A peptide as defined in claim 89 and two other peptides chosen from
the B to D peptides as defined in claim 89, or ii) the B peptide as
defined in claim 89 and two other peptides chosen from the A, C or
D peptides as defined in claim 89, or iii) the C peptide as defined
in claim 89 and two other peptides chosen from the A, B or D
peptides as defined in claim 89, or iv) the D peptide as defined in
claim 89 and two other peptides chosen from the A, B or C peptides
as defined in claim 89.
102. The fusion peptide according to claim 89, characterized in
that it comprises a fusion of the four A to D peptides.
103. The fusion peptide according to claim 89, characterized in
that it comprises a fusion of A to D peptides with a B peptide as
defined in claim 89 located at the N-terminus of the fusion which
is fused to a A peptide as defined in claim 89 which is fused to a
C peptide as defined in claim 89 which is fused to a D peptide as
defined in claim 89.
104. The fusion peptide according to claim 103, characterized in
that the A peptide has at least SEQ ID NO: 3 and at most SEQ ID NO:
19; and/or the B peptide has at least SEQ ID NO: 47 and at most SEQ
ID NO: 81; and/or the C peptide has at least SEQ ID NO: 129 and at
most SEQ ID NO: 147, SEQ ID NO: 153, SEQ ID NO: 162 or SEQ ID NO:
167; and/or the D peptide has at least SEQ ID NO: 176 and at most
SEQ ID NO: 188, SEQ ID NO: 192, SEQ ID NO: 193 or SEQ ID NO: 194,
SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ
ID NO: 205 or SEQ ID NO: 210.
105. The fusion peptide according to claim 104, characterized in
that the A peptide has the amino acid sequence defined in SEQ ID
NO: 3, the B peptide has the amino acid sequence defined in SEQ ID
NO: 47, the C peptide has the amino acid sequence defined in SEQ ID
NO: 129 and the D peptide has the amino acid sequence defined in
SEQ ID NO: 176.
106. The fusion peptide according to claim 105, characterized in
that it comprises the amino acid sequence defined in SEQ ID NO:
234.
107. A peptide composition characterized in that it comprises at
least a fusion peptide according to claim 89.
108. Nucleotide sequences coding for any one of the fusion peptides
as defined in claim 89.
109. An expression vector characterized in that it comprises a
nucleotide sequence according to claim 108, as well as the means
necessary for its expression.
110. Expression vector according to claim 109, characterized in
that said vector is a replication-defective adenoviral vector.
111. Expression vector according to claim 109, characterized in
that said vector is a MVA vaccinia vector.
112. A microorganism or a host cell transformed by an expression
vector as defined in claim 109.
113. Antibodies directed against at least one fusion peptide as
defined in claim 89.
114. A method for inhibiting, preventing, or treating an infection
caused by the hepatitis C virus or a HCV-associated disease or
pathologic conditions, in an animal, such as a human, the method
comprising the administration of a composition of claim 107 to the
animal.
115. A method for inhibiting, preventing, or treating an infection
caused by the hepatitis C virus or a HCV-associated disease or
pathologic conditions, in an animal, such as a human, the method
comprising the administration of at least one fusion peptide of
claim 89 to the animal.
116. A method for inhibiting, preventing, or treating an infection
caused by the hepatitis C virus or a HCV-associated disease or
pathologic conditions, in an animal, such as a human, the method
comprising the administration of at least one expression vector of
claim 109 to the animal.
117. A method for inhibiting, preventing, or treating an infection
caused by the hepatitis C virus or a HCV-associated disease or
pathologic conditions, in an animal, such as a human, the method
comprising the administration of at least one microorganism or host
cell of claim 112 to the animal.
118. A method for inhibiting, preventing, or treating an infection
caused by the hepatitis C virus or a HCV-associated disease or
pathologic conditions, in an animal, such as a human, the method
comprising the administration of at least one nucleotide sequence
of claim 108 to the animal.
119. A pharmaceutical composition, in particular vaccine,
containing as active ingredient at least one peptide composition
according to claim 107, in combination with a pharmaceutically
appropriate vehicle.
120. A pharmaceutical composition, in particular vaccine,
containing as active ingredient at least one of the antibodies as
defined in claim 113, in combination with a pharmaceutically
appropriate vehicle.
121. Pharmaceutical composition, in particular vaccine, containing
as active ingredient at least a fusion peptide according to claim
89, or at least a nucleotide sequence coding for said fusion
peptide, or at least a vector containing said nucleotide sequence
placed under the control of the elements necessary for a
constitutive and/or inducible expression of said fusion peptide in
a host cell, or at least a microorganism or a host cell transformed
by said vector, in combination with a pharmaceutically appropriate
vehicle.
122. A diagnostic composition for the detection and/or
quantification of the hepatitis C virus comprising at least one
peptide composition according to claim 107.
123. A diagnostic composition for the detection and/or
quantification of the hepatitis C virus comprising at least one
antibody according to claim 113.
124. A process for detection and/or quantification of the hepatitis
C virus in a biological sample taken from an individual capable of
being infected by said virus, such as plasma, serum or tissue,
characterized in that it comprises stages consisting of: bringing
said biological sample into contact with the antibodies according
to claim 113, under conditions allowing the formation of a complex
between the virus and the antibody, and detecting and/or
quantifying the formation of said complex by any appropriate
means.
125. A method for the in vitro diagnosis of the hepatitis C virus
in a biological sample, comprising contacting the sample with a
pharmaceutical composition, in particular vaccine, containing as
active ingredient at least a fusion peptide according to claim 89,
or at least a nucleotide sequence coding for said fusion peptide,
or at least a vector containing said nucleotide sequence placed
under the control of the elements necessary for a constitutive
and/or inducible expression of said fusion peptide in a host cell,
or at least a microorganism or a host cell transformed by said
vector, in combination with a pharmaceutically appropriate
vehicle.
126. A method for the in vitro diagnosis of the hepatitis C virus
in a biological sample, comprising contacting the sample with a
diagnostic composition as defined in claim 122 or 123.
127. A method of inducing or stimulating an immune response against
HCV in a host organism comprising administering to said organism
the peptide composition according to claim 107, so as to induce or
stimulate said immune response.
128. A method of inducing or stimulating an immune response against
HCV in a host organism comprising administering to said organism
the fusion peptide according to claim 89, so as to induce or
stimulate said immune response.
129. A method of inducing or stimulating an immune response against
HCV in a host organism comprising administering to said organism
the nucleotide sequence according to claim 108, so as to induce or
stimulate said immune response.
130. A method of inducing or stimulating an immune response against
HCV in a host organism comprising administering to said organism
the microorganism or host cell according to claim 112, so as to
induce or stimulate said immune response.
131. A method of inducing or stimulating an immune response against
HCV in a host organism comprising administering to said organism
the vector according to claim 109, so as to induce or stimulate
said immune response.
132. A method of inducing or stimulating an immune response against
HCV in a host organism comprising administering to said organism
the antibodies according to claim 113, so as to induce or stimulate
said immune response.
133. A method of inducing or stimulating an immune response against
HCV in a host organism comprising administering to said organism
the pharmaceutical composition according to any one of claims 119
to 121, so as to induce or stimulate said immune response.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of, and claims
priority from, U.S. Ser. No. 10/514,762, filed Nov. 16, 2004, which
claims priority under 35 U.S.C. 371 from PCT/FR03/001478, filed May
15, 2003, which claims priority from French application no.
02/06111, filed May 17, 2002. The contents of each of the prior
applications are incorporated herein by reference.
[0002] A subject of the present invention is novel peptide
compositions and their use in particular in the preparation of
pharmaceutical compositions active against the hepatitis C
virus.
[0003] A subject of the present invention is a peptide composition
useful in particular in prophylactic and therapeutic vaccination
directed against the hepatitis C virus.
[0004] Hepatitis C is the main cause of hepatitis acquired by
transfusion. Hepatitis C can also be transmitted by other
percutaneous routes, for example by injection of drugs by
intravenous route. The risk of contamination to health
professionals is moreover not negligible.
[0005] Hepatitis C differs from the other forms of liver diseases
associated with viruses, such as hepatitis A, B or D. Infections by
the hepatitis C virus (HCV) are mostly chronic, resulting in liver
diseases, such as hepatitis, cirrhosis and carcinoma in a large
number of cases (5 to 20%).
[0006] Although the risk of transmission of the virus by
transfusion has diminished due to the establishment of screening
tests in the 1990s, the frequency of hepatitis C remains high. By
way of example, a recent study indicates that today there are still
10,000 to 15,000 new cases of infection per year in France (S.
Deuffic et al., Hepatology 1999; 29: 1596-1601). At present,
approximately 170 million people world-wide are chronically
infected by HCV. Populations at high risk are chiefly hospital
staff and intravenous-drug users, but asymptomatic blood donors
exist who do not belong to these high-risk groups and in whom
circulating anti-HCV antibodies have been found. For the latter,
the infection route has not yet been identified.
[0007] HCV was the first hepatotropic virus isolated by means of
molecular biology techniques. The sequences of the viral genome
were cloned before the viral particle was visualized.
[0008] HCV belongs to a novel group of the Flaviviridae family, the
hepaciviruses. This is a virus with a single positive RNA strand,
of 9.5 kb, which is replicated by a copy of complementary RNA and
the translation product of which is a precursor of a single
polyprotein of approximately 3,000 amino acids. The 5' end of the
HCV genome corresponds to a non-translated region adjacent to the
genes which code for the structural proteins, the core protein of
the nucleocapsid, the two envelope glycoproteins, E1 and E2, and a
small protein called p7. The non-translated 5' region and the core
gene are relatively well preserved in the different genotypes. The
envelope proteins E1 and E2 are encoded by more variable regions
from one isolate to another. The protein p7 is an extremely
hydrophobic protein the function of which is not known. The 3' end
of the HCV genome contains the genes which code for the
non-structural proteins (NS2, NS3, NS4, NS5) and for a non-coding
3' region possessing a well-preserved domain (Major M E, Feinstone
S M, Hepatology, June 1997, 25(6): 1527-1538).
[0009] Therapy for the treatment of hepatitis C which is the
current focus of attention is a dual therapy using pegylated
interferon and ribavirin (Manns M P et al., The Lancet, 22 Sep.
2001, Vol. 358, 958-965). Whilst this therapy is particularly
effective in the case of patients infected by viral strains
belonging to genotypes 2 and 3, it only has a limited effect on
genotypes 1a, 1b and 4 (Manns M P, above).
[0010] It is therefore necessary to develop a vaccine composition
targeting these poorly-responsive genotypes as a priority.
[0011] Several studies today show that the control of an infection
caused by HCV, either naturally ("spontaneous resolution"), or
after treatment ("therapeutic resolution") is associated with the
induction or potentialization of cell-mediated immune responses
involving the T-CD4.sup.+ and T-CD8.sup.+ lymphocytes (CERNY A et
al., J. Clin. Invest., 95: 521-530 (1995)).
[0012] The object of vaccines based on the use of peptides is
generally to induce immune responses mediated by the T-CD4+ and/or
T-CD8+ lymphocytes.
[0013] The molecules of the major histocompatibility complex (MHC)
are described as class I or class II. Class I molecules are
expressed on virtually all of the nucleated cells and can be the
target of CD8.sup.+ cytotoxic T lymphocytes (CTLs). The CTLs
recognize the peptides or epitopes which are presented in
association with the MHC molecules of class I.
[0014] For example, the class I molecule HLA-A2.1 indicates that
the binding site of the peptide is created by bringing together the
domains .alpha..sub.1 and .alpha..sub.2 of the heavy chain of class
I (Bjorkman et al., Nature, 329: 506 (1987)).
[0015] Certain authors have concluded the immunogenic power of
peptide preparations on the basis of their good binding scores on
HLA molecules, as in the Patent Application WO01/21189.
[0016] Such a deduction is not evident and can lead: [0017] either
to the selection of peptides having no immunogenic power, although
having a high binding score, as demonstrated by the Applicant in
Example 1 of the present Application with the FLAT peptide, [0018]
or to the elimination of peptides which are in fact very
immunogenic, as shown with the CIN peptide in Brinster C. et al.
(Hepatology, Vol. 34, No 6, 2001, 1206-1217). In fact, although the
CIN peptide has an average, or even low (335), binding score, it is
nevertheless capable of inducing a strong response mediated by
cytotoxic T lymphocytes.
[0019] The Applicant has now unexpectedly found that a novel
peptide composition containing at least two peptides chosen from
the A to D peptides had a strong immunogenic power and had an
effect on the ability of the cells originating from patients
infected by viral strains of genotype 1a, 1b and 4 to induce
specific immune responses. These patients preferably, but not
limitatively, have an HLA of type HLA-A2.1.
[0020] Thus, a subject of the present invention is the peptide
compositions comprising at least two compounds chosen from: [0021]
an A peptide having at least the following amino acid sequence SEQ
ID NO: 1: [0022]
X.sub.1AX.sub.2QGYKVX.sub.3VLNPSVX.sub.4ATLX.sub.5FGX.sub.6YMSK-
AX.sub.7GX.sub.8,
[0023] in which X.sub.1 is Y or H, X.sub.2 is A, G or T, X.sub.3 is
R or L, X.sub.4 is A or T, X.sub.5 is G or S, X.sub.6 is A, T or V,
X.sub.7 is H or Y and X.sub.8 is I, T, M or V, [0024] a B peptide
having at least the following amino acid sequence SEQ ID NO: 45:
GX.sub.18X.sub.19X.sub.20X.sub.21X.sub.22X.sub.23TSLTGRDX.sub.24NX.sub.25-
X.sub.26X.sub.27GEX.sub.28QX.sub.29X.sub.30STAX.sub.31X.sub.32X.sub.33FLX.-
sub.34X.sub.35X.sub.36X.sub.37NGX.sub.38X.sub.39WTVX.sub.40
[0025] in which X.sub.18 is L or V, X.sub.19 is L or F, X.sub.20 is
G or S, X.sub.21 is C or T, X.sub.22 is I or V, X.sub.23 is I or V,
X.sub.24 is K, R or T, X.sub.25 is Q or E, X.sub.26 is V or N,
X.sub.27 is D, E or C, X.sub.28 is V or A, X.sub.29 is V, I, E or
M, X.sub.30 is L or V, X.sub.31 is T, K or A, X.sub.32 is Q or H,
X.sub.33 is S or T, X.sub.34 is A or G, X.sub.35 is T or S,
X.sub.36 is C or A, X.sub.37 is V, I or T, X.sub.38 is V or A,
X.sub.39 is C or M and X.sub.40 is Y or F, [0026] a C peptide
having at least the following amino acid sequence SEQ ID NO:
127:
[0027] SX.sub.47M
X.sub.48FTX.sub.49X.sub.50X.sub.51TSPLX.sub.52X.sub.53X.sub.54X.sub.55TLX-
.sub.56FNIX.sub.57GGWVAX.sub.58QX.sub.59
[0028] in which X.sub.47 is L or P, X.sub.48 is A or S, X.sub.49 is
A or S, X.sub.50 is A or S, X.sub.51 is I or V, X.sub.52 is T, S or
A, X.sub.53 is T or I, X.sub.54 is Q, S or G, X.sub.55 is N, Q, H,
S, Y or T, X.sub.56 is L or M, X.sub.57 is L or W, X.sub.58 is A or
S and X.sub.59 is L, P or I, and [0029] a D peptide having at least
the following amino acid sequence SEQ ID NO: 174:
[0030]
X.sub.69KX.sub.70ARX.sub.71IVX.sub.72PX.sub.73LGX.sub.74RVCEKX.sub-
.75ALX.sub.76X.sub.77VX.sub.78X.sub.79X.sub.80X.sub.81
[0031] in which X.sub.69 is R or Q, X.sub.70 is P or A, X.sub.71 is
L or F, X.sub.72 is F or Y, X.sub.73 is D or E, X.sub.74 is V or S,
X.sub.75 is M or R, X.sub.76 is Y or H, X.sub.77 is D or N,
X.sub.78 is V or I, X.sub.79 is S, T or K, X.sub.80 is T, K, I or N
and X.sub.81 is L or T, [0032] a B' epitope having the following
sequence SEQ ID NO: 213: [0033]
GX.sub.18X.sub.19X.sub.20X.sub.21X.sub.22X.sub.23TiSL
[0034] in which X.sub.18 is L or V, X.sub.19 is L or F, X.sub.20 is
G or S, X.sub.21 is C or T, X.sub.22 is I or V and X.sub.23 is I or
V, and [0035] a C' epitope having the following sequence SEQ ID NO:
221: [0036] SPLX.sub.52X.sub.53X.sub.54X.sub.55TL
[0037] in which X.sub.52 is T, S or A, X.sub.53 is T or I, X.sub.54
is Q, S or G and X.sub.55 is N, Q, H, S, Y or T,
[0038] as well as the pharmaceutical compositions containing them
and their use, in particular as vaccine, for the preparation of a
medicament intended for the inhibition or prevention of an
infection caused by the hepatitis C virus, and as a diagnostic
composition.
[0039] A particular subject of the invention is a peptide
composition as defined above, characterized in that it comprises at
least two peptides chosen from the A, B, C, D peptides.
[0040] A subject is also the particular A, B, C, D peptides, the
nucleotide sequences coding for said peptides and the
microorganisms or host cells cotransformed by these vectors.
[0041] Finally a subject is the antibodies directed against the
compositions of the invention and a process for detection and/or
quantification of the hepatitis C virus in a biological sample
using said antibodies.
[0042] The peptide compositions of the invention, having a strong
immunogenic power against the genotypes 1a, 1b and 4 of the
hepatitis C virus, therefore contain at least two peptides chosen
from the A to D peptides as defined above.
[0043] The A peptide is included in the non-structural protein 3
(NS3) between positions 1244 and 1274 of the polyprotein encoded by
the HCV virus.
[0044] The B peptide is also included in the protein NS3 between
positions 1038 and 1082 of the viral polyprotein.
[0045] The C peptide is included in the non-structural protein NS4
between positions 1789 and 1821 of said viral polyprotein.
[0046] As for the D peptide, it is included in the non-structural
protein NS5b between positions 2573 and 2601 of said viral
polyprotein.
[0047] Of course, by peptide is meant the peptide as such, as well
as its homologues having at least 60%, preferably at least 70%,
still more preferably at least 80%, better at least 90% and still
better 95% homology with the peptide of interest.
[0048] The A to D peptides were obtained from the strain HCV-JA
(Kato L., et al., (1990), Proc. Natl. Acad. Sci. USA, 87(24),
9524-9528) otherwise called the Shimotohno strain. They contain
known epitopes but, as indicated previously, these epitopes do not
necessarily have a high binding score.
[0049] The binding score is obtained by prediction, using software,
of the ability of known or potential epitope sequences (peptide
sequences) to bind to the HLA molecule of interest. This score can
be included in a range from negative values to the value of
approximately 1000 and by high binding score is meant a binding
score greater than or equal to 600.
[0050] On the other hand, unexpectedly, the combination of the A to
D peptides of the invention makes it possible to induce specific
cytotoxic T lymphocytes capable of a vigour and effectiveness
greater than that obtained with each of the peptides used
separately and/or each of the individual epitopes contained in
these peptides used alone or in combination as well as stimulating
a higher production of .quadrature.interferon.
[0051] Moreover, these peptides are capable of inducing specific T
lymphocytes having cross-reactivity.
[0052] According to one embodiment of the invention, the
compositions of the invention contain two peptides according to the
following combinations: A and B peptides, A and C peptides, A and D
peptides, B and C peptides, B and D peptides and C and D
peptides.
[0053] The preferred compositions contain the A and B, A and C, B
and D, and C and D peptides, the compositions containing the A and
B, C and D, and B and D peptides being more preferred.
[0054] According to another embodiment, the compositions of the
invention contain three peptides according to the following
combinations: A, B and D peptides, A, C and D peptides, and B, C
and D peptides, the compositions comprising the A, B and C, A, C
and D, and B, C and D peptides being particularly preferred.
[0055] According to yet another embodiment, the compositions of the
invention contain the four A, B, C, D peptides.
[0056] The A peptide in the compositions of the invention has at
least the amino acid sequence SEQ ID NO: 1 as described above.
[0057] According to one embodiment of the invention, the A peptide
has at most the 46 amino acids as described in the following
sequence SEQ ID NO: 2:
SX.sub.9X.sub.10VPX.sub.11X.sub.12X.sub.1AX.sub.2QGYKVX.sub.3VLNPSVX.sub-
.4ATLX.sub.5FGX.sub.6YMSKAX.sub.7GX.sub.8X.sub.13PX.sub.14X.sub.15X.sub.16-
X.sub.17GV,
[0058] in which X.sub.1 to X.sub.8 are as defined previously, and
X.sub.9 is T or N, X.sub.10 is K or R, X.sub.11 is A or V, X.sub.12
is A or E, X.sub.13 is E or D, X.sub.14 is N or S, X.sub.15 is I, L
or V, X.sub.16 is R or S and X.sub.17 is T or S.
[0059] In this case, the A peptide is situated between positions
1237 and 1282 of the viral polyprotein.
[0060] According to yet another embodiment, the A peptide is chosen
from the following peptides: [0061] the peptides having at least
sequence SEQ ID NO: 3, which corresponds to sequence SEQ ID NO: 1
in which X.sub.1 is Y, X.sub.2 is A, X.sub.3 is R, X.sub.4 is A,
X.sub.5 is G, X.sub.6 is A, X.sub.7 is H and X.sub.8 is I, and at
most sequence SEQ ID NO: 19 which corresponds to sequence SEQ ID
NO: 2 in which X.sub.1 to X.sub.8 are as defined for sequence SEQ
ID NO: 3 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12
is A, X.sub.13 is E, X.sub.14 is N, X.sub.15 is I, X.sub.16 is R
and X.sub.17 is T, [0062] the peptides having at least sequence SEQ
ID NO: 4 which corresponds to sequence SEQ ID NO: 1 in which
X.sub.1 is Y, X.sub.2 is A, X.sub.3 is L, X.sub.4 is A, X.sub.5 is
G, X.sub.6 is A, X.sub.7 is H and X.sub.8 is I, and at most a
sequence chosen from the following sequences: [0063] i) sequence
SEQ ID NO: 20 which corresponds to sequence SEQ ID NO: 2 in which
X.sub.1 to X.sub.8 are as defined for sequence SEQ ID NO: 4 and
X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13
is D, X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and X.sub.17 is
T, [0064] ii) sequence SEQ ID NO: 24 which corresponds to sequence
SEQ ID NO: 2 in which X.sub.1 to X.sub.8 are as defined for
sequence SEQ ID NO: 4 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is
A, X.sub.12 is A, X.sub.13 is E, X.sub.14 is N, X.sub.15 is I,
X.sub.16 is R and X.sub.17 is T, [0065] iii) sequence SEQ ID NO: 32
which corresponds to sequence SEQ ID NO: 2 in which X.sub.1 to
X.sub.8 are as defined for sequence SEQ ID NO: 4 and X.sub.9 is T,
X.sub.10 is R, X.sub.11 is A, X.sub.12 is A, X.sub.13 is D,
X.sub.14 is N, X.sub.15 is L, X.sub.16 is R and X.sub.17 is T, and
[0066] iv) sequence SEQ ID NO: 34 which corresponds to sequence SEQ
ID NO: 2 in which X.sub.1 to X.sub.8 are as defined for sequence
SEQ ID NO: 4 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is A,
X.sub.12 is A, X.sub.13 is D, X.sub.14 is N, X.sub.15 is V,
X.sub.16 is R and X.sub.17 is T, [0067] the peptides having at
least sequence SEQ ID NO: 5 which corresponds to sequence SEQ ID
NO: 1 in which X.sub.1 is Y, X.sub.2 is A, X.sub.3 is L, X.sub.4 is
A, X.sub.5 is G, X.sub.6 is A, X.sub.7 is H and X.sub.8 is V, and
at most a sequence chosen from the following sequences: [0068] i)
sequence SEQ ID NO: 21 which corresponds to sequence SEQ ID NO: 2
in which X.sub.1 to X.sub.8 are as defined for sequence SEQ ID NO:
5 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12 is A,
X.sub.13 is D, X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and
X.sub.17 is T, [0069] ii) sequence SEQ ID NO: 28 which corresponds
to sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8 are as defined
for sequence SEQ ID NO: 5 and X.sub.9 is N, X.sub.10 is K, X.sub.11
is V, X.sub.12 is E, X.sub.13 is D, X.sub.14 is N, X.sub.15 is I,
X.sub.16 is R and X.sub.17 is T, and [0070] iii) sequence SEQ ID
NO: 36 which corresponds to sequence SEQ ID NO: 2 in which X.sub.1
to X.sub.8 are as defined for sequence SEQ ID NO: 5 and X.sub.9 is
T, X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13 is D,
X.sub.14 is N, X.sub.15 is I, X.sub.16 is S and X.sub.17 is T,
[0071] the peptides having at least sequence SEQ ID NO: 6 which
corresponds to sequence SEQ ID NO: 1 in which X.sub.1 is Y, X.sub.2
is A, X.sub.3 is L, X.sub.4 is A, X.sub.5 is G, X.sub.6 is A,
X.sub.7 is H and X.sub.8 is T, and at most a sequence chosen from
the following sequences: [0072] i) sequence SEQ ID NO: 22 which
corresponds to sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8
are as defined for sequence SEQ ID NO: 6 and X.sub.9 is T, X.sub.10
is K, X.sub.11 is A, X.sub.12 is A, X.sub.13 is D, X.sub.14 is N,
X.sub.15 is I, X.sub.16 is R and X.sub.17 is T, [0073] ii) sequence
SEQ ID NO: 27 which corresponds to sequence SEQ ID NO: 2 in which
X.sub.1 to X.sub.8 are as defined for sequence SEQ ID NO: 6 and
X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13
is E, X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and X.sub.17 is
T, and [0074] iii) sequence SEQ ID NO: 41 which corresponds to
sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8 are as defined
for sequence SEQ ID NO: 6 and X.sub.9 is T, X.sub.10 is R, X.sub.11
is A, X.sub.12 is A, X.sub.13 is D, X.sub.14 is N, X.sub.15 is I,
X.sub.16 is R and X.sub.17 is T, [0075] the peptides having at
least sequence SEQ ID NO: 7 which corresponds to sequence SEQ ID
NO: 1 in which X.sub.1 is Y, X.sub.2 is A, X.sub.3 is L, X.sub.4 is
A, X.sub.5 is G, X.sub.6 is A, X.sub.7 is Y and X.sub.8 is T, and
at most a sequence chosen from the following sequences: [0076] i)
sequence SEQ ID NO: 23 which corresponds to sequence SEQ ID NO: 2
in which X.sub.1 to X.sub.8 are as defined for sequence SEQ ID NO:
7 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12 is A,
X.sub.13 is D, X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and
X.sub.17 is T, and [0077] ii) sequence SEQ ID NO: 37 which
corresponds to sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8
are as defined for sequence SEQ ID NO: 7 and X.sub.9 is T, X.sub.10
is K, X.sub.11 is A, X.sub.12 is A, X.sub.13 is D, X.sub.14 is N,
X.sub.15 is V, X.sub.16 is R and X.sub.17 is T, [0078] the peptides
having at least sequence SEQ ID NO: 8 which corresponds to sequence
SEQ ID NO: 1 in which X.sub.1 is Y, X.sub.2 is A, X.sub.3 is L,
X.sub.4 is A, X.sub.5 is S, X.sub.6 is A, X.sub.7 is H and X.sub.8
is T, and at most sequence SEQ ID NO: 25 which corresponds to
sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8 are as defined
for sequence SEQ ID NO: 8 and X.sub.9 is T, X.sub.10 is K, X.sub.11
is A, X.sub.12 is A, X.sub.13 is D, X.sub.14 is N, X.sub.15 is I,
X.sub.16 is R and X.sub.17 is T, [0079] the peptides having at
least sequence SEQ ID NO: 9 which corresponds to sequence SEQ ID
NO: 1 in which X.sub.1 is Y, X.sub.2 is A, X.sub.3 is L, X.sub.4 is
A, X.sub.5 is G, X.sub.6 is A, X.sub.7 is Y and X.sub.8 is I, and
at most sequence SEQ ID NO: 26 which corresponds to sequence SEQ ID
NO: 2 in which X.sub.1 to X.sub.8 are as defined for sequence SEQ
ID NO: 9 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12
is A, X.sub.13 is D, X.sub.14 is N, X.sub.15 is V, X.sub.16 is R
and X.sub.17 is T, [0080] the peptides having at least sequence SEQ
ID NO: 10 which corresponds to sequence SEQ ID NO: 1 in which
X.sub.1 is Y, X.sub.2 is A, X.sub.3 is L, X.sub.4 is T, X.sub.5 is
G, X.sub.6 is A, X.sub.7 is H and X.sub.8 is V, and at most
sequence SEQ ID NO: 29 which corresponds to sequence SEQ ID NO: 2
in which X.sub.1 to X.sub.8 are as defined for sequence SEQ ID NO:
10 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12 is A,
X.sub.13 is D, X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and
X.sub.17 is T, [0081] the peptides having at least sequence SEQ ID
NO: 11 which corresponds to sequence SEQ ID NO: 1 in which X.sub.1
is Y, X.sub.2 is G, X.sub.3 is L, X.sub.4 is A, X.sub.5 is G,
X.sub.6 is A, X.sub.7 is H and X.sub.8 is I, and at most sequence
SEQ ID NO: 30 which corresponds to sequence SEQ ID NO: 2 in which
X.sub.1 to X.sub.8 are as defined for sequence SEQ ID NO: 11 and
X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13
is D, X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and X.sub.17 is
T, [0082] the peptides having at least sequence SEQ ID NO: 12 which
corresponds to sequence SEQ ID NO: 1 in which X.sub.1 is Y, X.sub.2
is A, X.sub.3 is L, X.sub.4 is A, X.sub.5 is S, X.sub.6 is A,
X.sub.7 is H and X.sub.8 is I, and at most sequence SEQ ID NO: 31
which corresponds to sequence SEQ ID NO: 2 in which X.sub.1 to
X.sub.8 are as defined for sequence SEQ ID NO: 12 and X.sub.9 is T,
X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13 is D,
X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and X.sub.17 is T,
[0083] the peptides having at least sequence SEQ ID NO: 13 which
corresponds to sequence SEQ ID NO: 1 in which X.sub.1 is Y, X.sub.2
is T, X.sub.3 is L, X.sub.4 is A, X.sub.5 is G, X.sub.6 is A,
X.sub.7 is H and X.sub.8 is T, and at most sequence SEQ ID NO: 33
which corresponds to sequence SEQ ID NO: 2 in which X.sub.1 to
X.sub.8 are as defined for sequence SEQ ID NO: 13 and X.sub.9 is T,
X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13 is D,
X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and X.sub.17 is T,
[0084] the peptides having at least sequence SEQ ID NO: 14 which
corresponds to sequence SEQ ID NO: 1 in which X.sub.1 is Y, X.sub.2
is A, X.sub.3 is L, X.sub.4 is A, X.sub.5 is S, X.sub.6 is A,
X.sub.7 is H and X.sub.8 is V, and at most a sequence chosen from
the following sequences: [0085] i) sequence SEQ ID NO: 35 which
corresponds to sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8
are as defined for sequence SEQ ID NO: 14 and X.sub.9 is T,
X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13 is D,
X.sub.14 is S, X.sub.15 is I, X.sub.16 is R and X.sub.17 is T, and
[0086] ii) sequence SEQ ID NO: 39 which corresponds to sequence SEQ
ID NO: 2 in which X.sub.1 to X.sub.8 are as defined for sequence
SEQ ID NO: 14 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is A,
X.sub.12 is A, X.sub.13 is D, X.sub.14 is N, X.sub.15 is I,
X.sub.16 is R and X.sub.17 is T, [0087] the peptides having at
least sequence SEQ ID NO: 15 which corresponds to sequence SEQ ID
NO: 1 in which X.sub.1 is Y, X.sub.2 is T, X.sub.3 is L, X.sub.4 is
A, X.sub.5 is S, X.sub.6 is A, X.sub.7 is Y and X.sub.8 is M, and
at most sequence SEQ ID NO: 38 which corresponds to sequence SEQ ID
NO: 2 in which X.sub.1 to X.sub.8 are as defined for sequence SEQ
ID NO: 15 and X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12
is A, X.sub.13 is D, X.sub.14 is N, X.sub.15 is L, X.sub.16 is R
and X.sub.17 is T, [0088] the peptides having at least sequence SEQ
ID NO: 16 which corresponds to sequence SEQ ID NO: 1 in which
X.sub.1 is Y, X.sub.2 is A, X.sub.3 is L, X.sub.4 is A, X.sub.5 is
S, X.sub.6 is A, X.sub.7 is Y and X.sub.8 is V, and at most a
sequence chosen from the following sequences: [0089] i) sequence
SEQ ID NO: 40 which corresponds to sequence SEQ ID NO: 2 in which
X.sub.1 to X.sub.8 are as defined for sequence SEQ ID NO: 16 and
X.sub.9 is T, X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13
is D, X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and X.sub.17 is
T, and [0090] ii) sequence SEQ ID NO: 42 which corresponds to
sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8 are as defined
for sequence SEQ ID NO: 16 and X.sub.9 is T, X.sub.10 is R,
X.sub.11 is A, X.sub.12 is A, X.sub.13 is D, X.sub.14 is N,
X.sub.15 is I, X.sub.16 is R and X.sub.17 is T, [0091] the peptides
having at least sequence SEQ ID NO: 17 which corresponds to
sequence SEQ ID NO: 1 in which X.sub.1 is Y, X.sub.2 is A, X.sub.3
is L, X.sub.4 is A, X.sub.5 is G, X.sub.6 is T, X.sub.7 is Y and
X.sub.8 is T, and at most sequence SEQ ID NO: 43 which corresponds
to sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8 are as defined
for sequence SEQ ID NO: 17 and X.sub.9 is T, X.sub.10 is R,
X.sub.11 is A, X.sub.12 is A, X.sub.13 is D, X.sub.14 is N,
X.sub.15 is I, X.sub.16 is R and X.sub.17 is T, and [0092] the
peptides having at least sequence SEQ ID NO: 18 which corresponds
to sequence SEQ ID NO: 1 in which X.sub.1 is H, X.sub.2 is A,
X.sub.3 is L, X.sub.4 is A, X.sub.5 is G, X.sub.6 is V, X.sub.7 is
Y and X.sub.8 is I, and at most sequence SEQ ID NO: 44 which
corresponds to sequence SEQ ID NO: 2 in which X.sub.1 to X.sub.8
are as defined for sequence SEQ ID NO: 18 and X.sub.9 is T,
X.sub.10 is K, X.sub.11 is A, X.sub.12 is A, X.sub.13 is D,
X.sub.14 is N, X.sub.15 is I, X.sub.16 is R and X.sub.17 is S.
[0093] Preferably, the A peptide is chosen from the peptides of
sequences SEQ ID NO: 3 to 18, the peptide of sequence SEQ ID NO: 3
being particularly preferred.
[0094] The B peptide in the compositions of the invention has at
least the amino acid sequence SEQ ID NO: 45 as described above.
[0095] According to one embodiment, the B peptide has at most the
63 amino acids as described in the following sequence SEQ ID NO:
46:
AX.sub.41TX.sub.42YX.sub.43XQTRGX.sub.18X.sub.19X.sub.20X.sub.21X.sub.22-
X.sub.23TSLTGRDX.sub.24NX.sub.25X.sub.26X.sub.27GEX.sub.28QX.sub.29X.sub.3-
0STAX.sub.31X.sub.32X.sub.33FLX.sub.34X.sub.35X.sub.36X.sub.37NGX.sub.38X.-
sub.39WTVX.sub.40HGAGX.sub.45X.sub.46X.sub.47
[0096] in which X.sub.18 to X.sub.40 are as defined above and
X.sub.41 is P, S or H, X.sub.42 is A or T, X.sub.43 is S, A, T or
C, X.sub.44 is Q or R, X.sub.45 is S, T or A, X.sub.46 is K or R
and X.sub.47 is T or I.
[0097] In this case, the B peptide is situated between positions
1027 and 1089 of the viral polyprotein.
[0098] According to yet another embodiment the B peptide is chosen
from the following peptides: [0099] the peptides having at least
sequence SEQ ID NO: 47 which corresponds to sequence SEQ ID NO: 45
in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G, X.sub.21 is
C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q,
X.sub.26 is V, X.sub.27 is D, X.sub.28 is V, X.sub.29 is V,
X.sub.30 is L, X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S,
X.sub.34 is A, X.sub.35 is T, X.sub.36 is C, X.sub.37 is V,
X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and at most
sequence SEQ ID NO: 81 which corresponds to sequence SEQ ID NO: 46
in which X.sub.18 to X.sub.40 are as defined for sequence SEQ ID
NO: 47 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is
Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T, [0100] the
peptides having at least sequence SEQ ID NO: 48 which corresponds
to sequence SEQ ID NO: 45 in which X.sub.18 is L, X.sub.19 is L,
X.sub.20 is G, X.sub.21 is C, X.sub.22 is I, X.sub.23 is I,
X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E,
X.sub.28 is V, X.sub.29 is I, X.sub.30 is V, X.sub.31 is A,
X.sub.32 is Q, X.sub.33 is T, X.sub.34 is A, X.sub.35 is T,
X.sub.36 is C, X.sub.37 is I, X.sub.38 is V, X.sub.39 is C and
X.sub.40 is Y, and at most sequence SEQ ID NO: 82 which corresponds
to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40 are as
defined for sequence SEQ ID NO: 48 and X.sub.4 is P, X.sub.42 is A,
X.sub.43 is A, X.sub.44 is Q, X.sub.45 is T, X.sub.46 is R and
X.sub.47 is T, [0101] the peptides having at least sequence SEQ ID
NO: 49 which corresponds to sequence SEQ ID NO: 45 in which
X.sub.18 is L, X.sub.19 is L, X.sub.20 is G, X.sub.21 is C,
X.sub.22 is I, X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q,
X.sub.26 is V, X.sub.27 is E, X.sub.28 is V, X.sub.29 is I,
X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q, X.sub.33 is T,
X.sub.34 is A, X.sub.35 is T, X.sub.36 is C, X.sub.37 is I,
X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and at most
sequence SEQ ID NO: 83 which corresponds to sequence SEQ ID NO: 46
in which X.sub.18 to X.sub.40 are as defined for sequence SEQ ID
NO: 49 and X.sub.44 is P, X.sub.42 is A, X.sub.43 is T, X.sub.44 is
Q, X.sub.45 is T, X.sub.46 is R and X.sub.47 is T, [0102] the
peptides having at least sequence SEQ ID NO: 50 which corresponds
to sequence SEQ ID NO: 45 in which X.sub.18 is L, X.sub.19 is L,
X.sub.20 is G, X.sub.21 is C, X.sub.22 is I, X.sub.23 is I,
X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E,
X.sub.28 is V, X.sub.29 is V, X.sub.30 is V, X.sub.31 is T,
X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A, X.sub.35 is T,
X.sub.36 is C, X.sub.37 is V, X.sub.38 is V, X.sub.39 is C and
X.sub.40 is Y, and at most a sequence chosen from the following
sequences: [0103] i) sequence SEQ ID NO: 84 which corresponds to
sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40 are as defined
for sequence SEQ ID NO: 50 and X.sub.4 is P, X.sub.42 is A,
X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and
X.sub.47 is T, [0104] ii) sequence SEQ ID NO: 90 which corresponds
to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40 are as
defined for sequence SEQ ID NO: 50 and X.sub.41 is P, X.sub.42 is
A, X.sub.43 is S, X.sub.44 is R, X.sub.45 is S, X.sub.46 is K and
X.sub.47 is T, [0105] iii) sequence SEQ ID NO: 105 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 50 and X.sub.4 is P,
X.sub.42 is A, X.sub.43 is C, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0106] iv) sequence SEQ ID NO: 107
which corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to
X.sub.40 are as defined for sequence SEQ ID NO: 50 and X.sub.41 is
P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is T,
X.sub.46 is K and X.sub.47 is T, and [0107] v) sequence SEQ ID NO:
116 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 50 and X.sub.41
is P, X.sub.42 is T, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is T,
X.sub.46 is K and X.sub.47 is T, [0108] the peptides having at
least sequence SEQ ID NO: 51 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is R,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most a sequence chosen from the following sequences: [0109] i)
sequence SEQ ID NO: 85 which corresponds to sequence SEQ ID NO: 46
in which X.sub.18 to X.sub.40 are as defined for sequence SEQ ID
NO: 51 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is
Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T, and [0110] ii)
sequence SEQ ID NO: 124 which corresponds to sequence SEQ ID NO: 46
in which X.sub.18 to X.sub.40 are as defined for sequence SEQ ID
NO: 51 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is
Q, X.sub.45 is A, X.sub.46 is K and X.sub.47 is T, [0111] the
peptides having at least sequence SEQ ID NO: 52 which corresponds
to sequence SEQ ID NO: 45 in which X.sub.18 is L, X.sub.19 is L,
X.sub.20 is G, X.sub.21 is C, X.sub.22 is I, X.sub.23 is I,
X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E,
X.sub.28 is V, X.sub.29 is V, X.sub.30 is V, X.sub.31 is T,
X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A, X.sub.35 is T,
X.sub.36 is C, X.sub.37 is I, X.sub.38 is V, X.sub.39 is C and
X.sub.40 is Y, and at most a sequence chosen from the following
sequences: [0112] i) sequence SEQ ID NO: 86 which corresponds to
sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40 are as defined
for sequence SEQ ID NO: 52 and X.sub.41 is P, X.sub.42 is A,
X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and
X.sub.47 is T, and [0113] ii) sequence SEQ ID NO: 120 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 52 and X.sub.41 is P,
X.sub.42 is A, X.sub.43 is A, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0114] the peptides having at
least sequence SEQ ID NO: 53 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.58 is L, X.sub.19 is F, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 87 which corresponds to sequence SEQ ID
NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 53 and X.sub.4, is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T,
[0115] the peptides having at least sequence SEQ ID NO: 54 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is V,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is V,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is I, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is Y, and at most a sequence chosen from
the following sequences: [0116] i) sequence SEQ ID NO: 88 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 54 and X.sub.4, is P,
X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, and [0117] ii) sequence SEQ ID NO:
117 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 54 and X.sub.41
is S, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0118] the peptides having at
least sequence SEQ ID NO: 55 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is A, X.sub.39 is C and X.sub.40 is F, and
at most sequence SEQ ID NO: 89 which corresponds to sequence SEQ ID
NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 55 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T,
[0119] the peptides having at least sequence SEQ ID NO: 56 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is I, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is F, and at most a sequence chosen from
the following sequences: [0120] i) sequence SEQ ID NO: 91 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 56 and X.sub.41 is P,
X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is A,
X.sub.46 is K and X.sub.47 is T, and [0121] ii) sequence SEQ ID NO:
92 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to
X.sub.40 are as defined for sequence SEQ ID NO: 56 and X.sub.41 is
P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0122] the peptides having at
least sequence SEQ ID NO: 57 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is V, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is I, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 93 which corresponds to sequence SEQ ID
NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 57 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.4 is S, X.sub.46 is K and X.sub.47 is T,
[0123] the peptides having at least sequence SEQ ID NO: 58 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is V, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is F, and at most a sequence chosen from
the following sequences: [0124] i) sequence SEQ ID NO: 94 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 58 and X.sub.41 is P,
X.sub.42 is A, X.sub.43 is A, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, and [0125] ii) sequence SEQ ID NO:
110 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 58 and X.sub.41
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is T,
X.sub.46 is K and X.sub.47 is T, [0126] the peptides having at
least sequence SEQ ID NO: 59 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is V, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is A, X.sub.39 is C and X.sub.40 is F, and
at most sequence SEQ ID NO: 95 which corresponds to sequence SEQ ID
NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 59 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.4 is S, X.sub.46 is K and X.sub.47 is T,
[0127] the peptides having at least sequence SEQ ID NO: 60 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is V,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is V, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is F, and at most a sequence chosen from
the following sequences: [0128] i) sequence SEQ ID NO: 96 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 60 and X.sub.4, is P,
X.sub.42 is A, X.sub.43 is A, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, and [0129] ii) sequence SEQ ID NO:
115 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 60 and X.sub.41
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0130] the peptides having at
least sequence SEQ ID NO: 61 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is D, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is L, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is I, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 97 which corresponds to sequence SEQ ID
NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 61 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T,
[0131] the peptides having at least sequence SEQ ID NO: 62 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is V, X.sub.38 is A,
X.sub.39 is C and X.sub.40 is Y, and at most a sequence chosen from
the following sequences: [0132] i) sequence SEQ ID NO: 98 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 62 and X.sub.41 is P,
X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, and [0133] ii) sequence SEQ ID NO:
109 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 62 and X
.sub.41 is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45
is T, X.sub.46 is K and X.sub.47 is T, [0134] the peptides having
at least sequence SEQ ID NO: 63 which corresponds to sequence SEQ
ID NO: 45 in which X.sub.18 is V, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is H,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is I, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 99 which corresponds to sequence SEQ ID
NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 63 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T,
[0135] the peptides having at least sequence SEQ ID NO: 64 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is I, X.sub.38 is A,
X.sub.39 is C and X.sub.40 is Y, and at most sequence SEQ ID NO:
100 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 64 and X.sub.41
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0136] the peptides having at
least sequence SEQ ID NO: 65 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is S, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 101 which corresponds to sequence SEQ
ID NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 65 and X.sub.4, is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T,
[0137] the peptides having at least sequence SEQ ID NO: 66 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is T, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is Y, and at most sequence SEQ ID NO:
102 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 66 and X.sub.4,
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0138] the peptides having at
least sequence SEQ ID NO: 67 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is V, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 103 which corresponds to sequence SEQ
ID NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 67 and X.sub.4, is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T,
[0139] the peptides having at least sequence SEQ ID NO: 68 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is K, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is V, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is Y, and at most sequence SEQ ID NO:
104 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 68 and X.sub.4,
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0140] the peptides having at
least sequence SEQ ID NO: 69 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is K, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is A, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 106 which corresponds to sequence SEQ
ID NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 69 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T,
[0141] the peptides having at least sequence SEQ ID NO: 70 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is T, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is V, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is F, and at most sequence SEQ ID NO:
108 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 70 and X.sub.41
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0142] the peptides having at
least sequence SEQ ID NO: 71 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is E, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 111 which corresponds to sequence SEQ
ID NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 71 and X.sub.41 is P, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is I,
[0143] the peptides having at least sequence SEQ ID NO: 72 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is F, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is V, X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is A, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is V, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is Y, and at most sequence SEQ ID NO:
112 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 72 and X.sub.4
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0144] the peptides having at
least sequence SEQ ID NO: 73 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is F, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is I, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 113 which corresponds to sequence SEQ
ID NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 73 and X.sub.4, is P, X.sub.42 is A, X.sub.43 is S, 4 is
Q, X.sub.45 is S, X.sub.46 is K and X.sub.47 is T, [0145] the
peptides having at least sequence SEQ ID NO: 74 which corresponds
to sequence SEQ ID NO: 45 in which X.sub.18 is L, X.sub.19 is F,
X.sub.20 is G, X.sub.21 is C, X.sub.22 is I, X.sub.23 is V,
X.sub.24 is K, X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E,
X.sub.28 is A, X.sub.29 is V, X.sub.30 is V, X.sub.31 is T,
X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A, X.sub.35 is T,
X.sub.36 is C, X.sub.37 is V, X.sub.38 is A, X.sub.39 is C and
X.sub.40 is Y, and at most sequence SEQ ID NO: 114 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 74 and X.sub.4i is P,
X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.4 is S, X.sub.46
is K and X.sub.47 is T, [0146] the peptides having at least
sequence SEQ ID NO: 75 which corresponds to sequence SEQ ID NO: 45
in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G, X.sub.21 is
C, X.sub.22 is I, X.sub.23 is V, X.sub.24 is K, X.sub.25 is Q,
X.sub.26 is V, X.sub.27 is E, X.sub.28 is V, X.sub.29 is V,
X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S,
X.sub.34 is A, X.sub.35 is T, X.sub.36 is C, X.sub.37 is V,
X.sub.38 is V, X.sub.39 is C and X.sub.40 is F, and at most a
sequence chosen from the following sequences: [0147] i) sequence
SEQ ID NO: 118 which corresponds to sequence SEQ ID NO: 46 in which
X.sub.18 to X.sub.40 are as defined for sequence SEQ ID NO: 75 and
X.sub.4, is S, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.4
is S, X.sub.46 is K and X.sub.47 is T, and [0148] ii) sequence SEQ
ID NO: 119 which corresponds to sequence SEQ ID NO: 46 in which
X.sub.18 to X.sub.40 are as defined for sequence SEQ ID NO: 75 and
X.sub.41 is P, X.sub.42 is A, X.sub.43 is A, X.sub.44 is Q,
X.sub.45 is S, X.sub.46 is K and X.sub.47 is T, [0149] the peptides
having at least sequence SEQ ID. NO: 76 which corresponds to
sequence SEQ ID NO: 45 in which X.sub.18 is L, X.sub.19 is F,
X.sub.20 is G, X.sub.21 is C, X.sub.22 is I, X.sub.23 is I,
X.sub.24 is R, X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E,
X.sub.28 is V, X.sub.29 is V, X.sub.30 is V, X.sub.31 is T,
X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A, X.sub.35 is T,
X.sub.36 is C, X.sub.37 is I, X.sub.38 is V, X.sub.39 is C and
X.sub.40 is Y, and at most sequence SEQ ID NO: 121 which
corresponds to sequence SEQ ID NO: 46 in which X.sub.18 to X.sub.40
are as defined for sequence SEQ ID NO: 76 and X.sub.4, is P,
X.sub.42 is A, X.sub.43 is A, X.sub.44 is Q, X.sub.45 is S,
X.sub.46 is K and X.sub.47 is T, [0150] the peptides having at
least sequence SEQ ID NO: 77 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is F, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is K,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is V, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is I, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 122 which corresponds to sequence SEQ
ID NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 77 and X.sub.4, is P, X.sub.42 is A, X.sub.43 is A,
X.sub.44 is Q, X.sub.4 is S, X.sub.46 is K and X.sub.47 is T,
[0151] the peptides having at least sequence SEQ ID NO: 78 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is L, X.sub.20 is G, X.sub.21 is C, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is R, X.sub.25 is Q, X.sub.26 is V,
X.sub.27 is E, X.sub.28 is V, X.sub.29 is V, X.sub.30 is V,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is A,
X.sub.35 is T, X.sub.36 is C, X.sub.37 is I, X.sub.38 is V,
X.sub.39 is C and X.sub.40 is F, and at most sequence SEQ ID NO:
123 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 78 and X.sub.41
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.4 is S,
X.sub.46 is K and X.sub.47 is T, [0152] the peptides having at
least sequence SEQ ID NO: 79 which corresponds to sequence SEQ ID
NO: 45 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G,
X.sub.21 is C, X.sub.22 is I, X.sub.23 is I, X.sub.24 is R,
X.sub.25 is Q, X.sub.26 is V, X.sub.27 is E, X.sub.28 is V,
X.sub.29 is M, X.sub.30 is V, X.sub.31 is T, X.sub.32 is Q,
X.sub.33 is S, X.sub.34 is A, X.sub.35 is T, X.sub.36 is C,
X.sub.37 is V, X.sub.38 is V, X.sub.39 is C and X.sub.40 is Y, and
at most sequence SEQ ID NO: 125 which corresponds to sequence SEQ
ID NO: 46 in which X.sub.18 to X.sub.40 are as defined for sequence
SEQ ID NO: 79 and X.sub.41 is H, X.sub.42 is A, X.sub.43 is S,
X.sub.44 is Q, X.sub.4 is S, X.sub.46 is K and X.sub.47 is T, and
[0153] the peptides having at least sequence SEQ ID NO: 80 which
corresponds to sequence SEQ ID NO: 45 in which X.sub.18 is L,
X.sub.19 is F, X.sub.20 is S, X.sub.21 is T, X.sub.22 is I,
X.sub.23 is I, X.sub.24 is T, X.sub.25 is E, X.sub.26 is N,
X.sub.27 is C, X.sub.28 is V, X.sub.29 is V, X.sub.30 is L,
X.sub.31 is T, X.sub.32 is Q, X.sub.33 is S, X.sub.34 is G,
X.sub.35 is T, X.sub.36 is A, X.sub.37 is V, X.sub.38 is V,
X.sub.39 is M and X.sub.40 is Y, and at most sequence SEQ ID NO:
126 which corresponds to sequence SEQ ID NO: 46 in which X.sub.18
to X.sub.40 are as defined for sequence SEQ ID NO: 80 and X.sub.4,
is P, X.sub.42 is A, X.sub.43 is S, X.sub.44 is Q, X.sub.45 is A,
X.sub.46 is K and X.sub.47 is T.
[0154] Preferably, the B peptide is chosen from the peptides of
sequences SEQ ID NO: 47 to 80, the peptide of sequence SEQ ID NO:
47 being particularly preferred.
[0155] The C peptide in the compositions of the invention has at
least the amino acid sequence SEQ ID NO: 127 as described
previously.
[0156] According to one embodiment, the C peptide has at most the
57 amino acids as described in the following sequence SEQ ID NO:
128: NFIX.sub.60GX.sub.61 QYLAX.sub.62LSTLPGNX.sub.63AX.sub.65
SX.sub.47M.sub.8FTX.sub.49X.sub.50X.sub.5
TSPLX.sub.52X.sub.53X.sub.54X.sub.55TLX.sub.56FNIX.sub.57GGWVAX.sub.58QX.-
sub.59X.sub.66X.sub.67X.sub.68
[0157] in which X.sub.47 to X.sub.59 are as defined above and
X.sub.60 is T or S, X.sub.61 is I, T or V, X.sub.62 is G or A,
X.sub.63 is P or L, X.sub.64 is I or M, X.sub.65 is A, V or R,
X.sub.66 is A or R, X.sub.67 is P, A or D and X.sub.68 is P, A or
S.
[0158] In this case, the C peptide is situated between positions
1767 and 1823 of the viral polyprotein.
[0159] According to another embodiment, the C peptide is chosen
from the following peptides: [0160] the peptides having at least
sequence SEQ ID NO: 129 which corresponds to sequence SEQ ID NO:
127 in which X.sub.47 is L, X.sub.48 is A, X.sub.49 is A, X.sub.50
is S, X.sub.51 is I, X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q,
X.sub.55 is N, X.sub.56 is L, X.sub.57 is L, X.sub.58 is A and
X.sub.59 is L and at most a sequence chosen from the following
sequences: [0161] i) sequence SEQ ID NO: 147 which corresponds to
sequence SEQ ID NO: 128 in which X.sub.47 to X.sub.59 are as
defined for sequence SEQ ID NO: 129 and X.sub.60 is T, X.sub.61 is
I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A,
X.sub.66 is A, X.sub.67 is P and X.sub.68 is P, [0162] ii) sequence
SEQ ID NO: 153 which corresponds to sequence SEQ ID NO: 128 in
which X.sub.47 to X.sub.59 are as defined for sequence SEQ ID NO:
129 and X.sub.60 is S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P,
X.sub.64 is I, X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and
X.sub.68 is P, [0163] iii) sequence SEQ ID NO: 162 which
corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 129 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is A, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is A, and
[0164] iv) sequence SEQ ID NO: 167 which corresponds to sequence
SEQ ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 129 and X.sub.60 is S, X.sub.61 is V, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0165] the peptides having at
least sequence SEQ ID NO: 130 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.48 is A, X.sub.49 is A,
X.sub.50 is A, X.sub.51 is V, X.sub.52 is T, X.sub.53 is T,
X.sub.54 is S, X.sub.55 is Q, X.sub.56 is L, X.sub.57 is L,
X.sub.58 is A and X.sub.59 is L and at most a sequence chosen from
the following sequences: [0166] i) sequence SEQ ID NO: 148 which
corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 130 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is A and X.sub.68 is P, and
[0167] ii) sequence SEQ ID NO: 150 which corresponds to sequence
SEQ ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 130 and X.sub.60 is S, X.sub.61 is T, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is A and X.sub.68 is P, [0168] the peptides having at
least sequence SEQ ID NO: 131 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.48 is A, X.sub.49 is A,
X.sub.50 is A, X.sub.5, is V, X.sub.52 is T, X.sub.53 is T,
X.sub.54 is G, X.sub.55 is Q, X.sub.56 is L, X.sub.57 is L,
X.sub.58 is A and X.sub.59 is L and at most sequence SEQ ID NO: 149
which corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 131 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is A and X.sub.68 is P
[0169] the peptides having at least sequence SEQ ID NO: 132 which
corresponds to sequence SEQ ID NO: 127 in which X.sub.47 is L,
X.sub.49 is A, X.sub.49 is A, X.sub.50 is S, X.sub.51 is I,
X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q, X.sub.55 is H,
X.sub.56 is L, X.sub.57 is L, X.sub.58 is A and X.sub.59 is L and
at most a sequence chosen from the following sequences: [0170] i)
sequence SEQ ID NO: 151 which corresponds to sequence SEQ ID NO:
128 in which X.sub.47 to X.sub.59 are as defined for sequence SEQ
ID NO: 132 and X.sub.60 is S, X.sub.61 is I, X.sub.62 is G,
X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0171] ii) sequence SEQ ID NO: 155
which corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 132 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is V, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P,
[0172] iii) sequence SEQ ID NO: 168 which corresponds to sequence
SEQ ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 132 and X.sub.60 is S, X.sub.61 is V, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, and [0173] iv) sequence SEQ ID NO:
171 which corresponds to sequence SEQ ID NO: 128 in which X.sub.47
to X.sub.59 are as defined for sequence SEQ ID NO: 132 and X.sub.60
is S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is L, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P,
[0174] the peptides having at least sequence SEQ ID NO: 133 which
corresponds to sequence SEQ ID NO: 127 in which X.sub.47 is L,
X.sub.48 is A, X.sub.49 is A, X.sub.50 is S, X.sub.51 is I,
X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q, X.sub.55 is S,
X.sub.56 is L, X.sub.57 is L, X.sub.58 is A and X.sub.59 is L and
at most sequence SEQ ID NO: 152 which corresponds to sequence SEQ
ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 133 and X.sub.60 is S, X.sub.61 is I, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0175] the peptides having at
least sequence SEQ ID NO: 134 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.48 is A, X.sub.49 is A,
X.sub.50 is S, X.sub.51 is I, X.sub.52 is T, X.sub.53 is T,
X.sub.54 is Q, X.sub.55 is Y, X.sub.56 is L, X.sub.57 is L,
X.sub.58 is A and X.sub.59 is L and at most a sequence chosen from
the following sequences: [0176] i) sequence SEQ ID NO: 154 which
corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 134 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P, and
[0177] ii) sequence SEQ ID NO: 169 which corresponds to sequence
SEQ ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 134 and X.sub.60 is S, X.sub.61 is V, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0178] the peptides having at
least sequence SEQ ID NO: 135 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.548 is A, X.sub.49 is A,
X.sub.50 is S, X.sub.51 is V, X.sub.52 is T, X.sub.53 is T,
X.sub.54 is Q, X.sub.55 is N, X.sub.56 is L, X.sub.57 is L,
X.sub.58 is A and X.sub.59 is L and at most sequence SEQ ID NO: 156
which corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 135 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P,
[0179] the peptides having at least sequence SEQ ID NO: 136 which
corresponds to sequence SEQ ID NO: 127 in which X.sub.47 is L,
X.sub.48 is A, X.sub.49 is A, X.sub.50 is S, X.sub.51 is V,
X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q, X.sub.55 is S,
X.sub.56 is L, X.sub.57 is L, X.sub.58 is A and X.sub.59 is L and
at most a sequence chosen from the following sequences: [0180] i)
sequence SEQ ID NO: 157 which corresponds to sequence SEQ ID NO:
128 in which X.sub.47 to X.sub.59 are as defined for sequence SEQ
ID NO: 136 and X.sub.60 is S, X.sub.61 is I, X.sub.62 is G,
X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, and [0181] ii) sequence SEQ ID NO:
170 which corresponds to sequence SEQ ID NO: 128 in which X.sub.47
to X.sub.59 are as defined for sequence SEQ ID NO: 136 and X.sub.60
is S, X.sub.61 is V, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P,
[0182] the peptides having at least sequence SEQ ID NO: 137 which
corresponds to sequence SEQ ID NO: 127 in which X.sub.47 is L,
X.sub.48 is A, X.sub.49 is A, X.sub.50 is S, X.sub.51 is I,
X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q, X.sub.55 is T,
X.sub.56 is M, X.sub.57 is L, X.sub.58 is A and X.sub.59 is L and
at most sequence SEQ ID NO: 158 which corresponds to sequence SEQ
ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 137 and X.sub.60 is S, X.sub.61 is I, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0183] the peptides having at
least sequence SEQ ID NO: 138 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.48 is A, X.sub.49 is A,
X.sub.50 is S, X.sub.51 is V, X.sub.52 is T, X.sub.53 is T,
X.sub.54 is Q, X.sub.55 is Y, X.sub.56 is L, X.sub.57 is L,
X.sub.58 is A and X.sub.59 is I and at most sequence SEQ ID NO: 159
which corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 138 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P,
[0184] the peptides having at least sequence SEQ ID NO: 139 which
corresponds to sequence SEQ ID NO: 127 in which X.sub.47 is P,
X.sub.48 is A, X.sub.49 is A, X.sub.50 is S, X.sub.51 is I,
X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q, X.sub.55 is H,
X.sub.56 is L, X.sub.57 is L, X.sub.58 is A and X.sub.59 is L and
at most sequence SEQ ID NO: 160 which corresponds to sequence SEQ
ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 139 and X.sub.60 is S, X.sub.61 is I, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is R, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0185] the peptides having at
least sequence SEQ ID NO: 140 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.48 is A, X.sub.49 is A,
X.sub.50 is S, X.sub.51 is I, X.sub.52 is T, X.sub.53 is I,
X.sub.54 is Q, X.sub.55 is H, X.sub.56 is L, X.sub.57 is L,
X.sub.58 is A and X.sub.59 is P and at most sequence SEQ ID NO: 161
which corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 140 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P,
[0186] the peptides having at least sequence SEQ ID NO: 141 which
corresponds to sequence SEQ ID NO: 127 in which X.sub.47 is L,
X.sub.48 is A, X.sub.49 is S, X.sub.50 is S, X.sub.51 is I,
X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q, X.sub.55 is S,
X.sub.56 is L, X.sub.57 is L, X.sub.58 is A and X.sub.59 is L and
at most sequence SEQ ID NO: 163 which corresponds to sequence SEQ
ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 141 and X.sub.60 is S, X.sub.61 is I, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0187] the peptides having at
least sequence SEQ ID NO: 142 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.48 is A, X.sub.49 is A,
X.sub.50 is S, X.sub.51 is I, X.sub.52 is S, X.sub.53 is T,
X.sub.54 is Q, X.sub.55 is N, X.sub.56 is L, X.sub.57 is W,
X.sub.58 is A and X.sub.59 is L and at most sequence SEQ ID NO: 164
which corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 142 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P,
[0188] the peptides having at least sequence SEQ ID NO: 143 which
corresponds to sequence SEQ ID NO: 127 in which X.sub.47 is L,
X.sub.48 is A, X.sub.49 is A, X.sub.50 is S, X.sub.51 is I,
X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q, X.sub.55 is N,
X.sub.56 is M, X.sub.57 is L, X.sub.58 is A and X.sub.59 is L and
at most sequence SEQ ID NO: 165 which corresponds to sequence SEQ
ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 143 and X.sub.60 is S, X.sub.61 is I, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0189] the peptides having at
least sequence SEQ ID NO: 144 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.48 is A, X.sub.49 is A,
X.sub.50 is S, X.sub.51 is I, X.sub.52 is T, X.sub.53 is T,
X.sub.54 is Q, X.sub.55 is H, X.sub.56 is M, X.sub.57 is L,
X.sub.58 is A and X.sub.59 is L and at most sequence SEQ ID NO: 166
which corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 144 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is M,
X.sub.65 is A, X.sub.66 is A, X.sub.67 is P and X.sub.68 is P,
[0190] the peptides having at least sequence SEQ ID NO: 145 which
corresponds to sequence SEQ ID NO: 127 in which X.sub.47 is L,
X.sub.49 is A, X.sub.49 is A, X.sub.50 is S, X.sub.51 is I,
X.sub.52 is A, X.sub.53 is T, X.sub.54 is Q, X.sub.55 is Y,
X.sub.56 is L, X.sub.57 is L, X.sub.58 is A and X.sub.59 is L and
at most sequence SEQ ID NO: 172 which corresponds to sequence SEQ
ID NO: 128 in which X.sub.47 to X.sub.59 are as defined for
sequence SEQ ID NO: 145 and X.sub.60 is S, X.sub.61 is I, X.sub.62
is G, X.sub.63 is P, X.sub.64 is I, X.sub.65 is A, X.sub.66 is A,
X.sub.67 is P and X.sub.68 is P, [0191] the peptides having at
least sequence SEQ ID NO: 146 which corresponds to sequence SEQ ID
NO: 127 in which X.sub.47 is L, X.sub.48 is S, X.sub.49 is A,
X.sub.50 is A, X.sub.5, is V, X.sub.52 is T, X.sub.53 is T,
X.sub.54 is Q, X.sub.55 is Q, X.sub.56 is L, X.sub.57 is L,
X.sub.58 is S and X.sub.59 is I and at most sequence SEQ ID NO: 173
which corresponds to sequence SEQ ID NO: 128 in which X.sub.47 to
X.sub.59 are as defined for sequence SEQ ID NO: 146 and X.sub.60 is
S, X.sub.61 is I, X.sub.62 is G, X.sub.63 is P, X.sub.64 is I,
X.sub.65 is A, X.sub.66 is R, X.sub.67 is D and X.sub.68 is S.
[0192] Preferably, the C peptide is chosen from the peptides of
sequences SEQ ID NO: 129 to 146, the peptide of sequence SEQ ID NO:
129 being particularly preferred.
[0193] The D peptide in the compositions of the invention has at
least the amino acid sequence SEQ ID NO: 174 as described
above.
[0194] According to one embodiment of the invention, the D peptide
has at most the 44 amino acids as described in the following
sequence SEQ ID NO: 175:
KGGX.sub.69KX.sub.70ARX.sub.71
IVX.sub.72PX.sub.73LGX.sub.74RVCEKX.sub.75ALX.sub.76X.sub.77VX.sub.78X.su-
b.79X.sub.80X.sub.81X.sub.82X.sub.83X.sub.84
VMGX.sub.85X.sub.86YX.sub.87X.sub.88Q
[0195] in which X.sub.69 to X.sub.8, are as defined above and
X.sub.82 is P or A, X.sub.83 is Q, L, H, R, K or P, X.sub.84 is A,
T, V or P, X.sub.85 is P, S or A, X.sub.86 is S or A, X.sub.87 is G
or R and X.sub.88 is F or C.
[0196] In this case, the D peptide is situated between positions
2570 and 2613 of the viral polyprotein.
[0197] According to a further embodiment, the D peptide is chosen
from the following peptides: [0198] the peptides having at least
sequence SEQ ID NO: 176 which corresponds to sequence SEQ ID NO:
174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is L, X.sub.72
is F, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M, X.sub.76 is Y,
X.sub.77 is D, X.sub.78 is V, X.sub.79 is S, X.sub.80 is T and
X.sub.81 is L, and at most a sequence chosen from: [0199] i)
sequence SEQ ID NO: 188 which corresponds to sequence SEQ ID NO:
175 in which X.sub.69 to X.sub.8, are as defined for sequence SEQ
ID NO: 176 and X.sub.82 is P, X.sub.83 is Q, X.sub.84 is A,
X.sub.85 is P, X.sub.86 is S, X.sub.87 is G and X.sub.88 is F,
[0200] ii) sequence SEQ ID NO: 192 which corresponds to sequence
SEQ ID NO: 175 in which X.sub.69 to X.sub.8, are as defined for
sequence SEQ ID NO: 176 and X.sub.82 is P, X.sub.83 is Q, X.sub.84
is A, X.sub.85 is S, X.sub.86 is S, X.sub.87 is G and X.sub.88 is
F, [0201] iii) sequence SEQ ID NO: 193 which corresponds to
sequence SEQ ID NO: 175 in which X.sub.69 to X.sub.81 are as
defined for sequence SEQ ID NO: 176 and X.sub.82 is P, X.sub.83 is
Q, X.sub.84 is A, X.sub.85 is A, X.sub.86 is S, X.sub.87 is G and
X.sub.88 is F, [0202] iv) sequence SEQ ID NO: 194 which corresponds
to sequence SEQ ID NO: 175 in which X.sub.69 to X.sub.81 are as
defined for sequence SEQ ID NO: 176 and X.sub.82 is P, X.sub.83 is
Q, X.sub.84 is V, X.sub.85 is S, X.sub.86 is S, X.sub.87 is G and
X.sub.88 is F, [0203] v) sequence SEQ ID NO: 201 which corresponds
to sequence SEQ ID NO: 175 in which X.sub.69 to X.sub.81 are as
defined for sequence SEQ ID NO: 176 and X.sub.82 is P, X.sub.83 is
R, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S, X.sub.87 is G and
X.sub.88 is C, [0204] vi) sequence SEQ ID NO: 202 which corresponds
to sequence SEQ ID NO: 175 in which X.sub.69 to X.sub.8, are as
defined for sequence SEQ ID NO: 176 and X.sub.82 is P, X.sub.83 is
Q, X.sub.84 is P, X.sub.85 is S, X.sub.86 is S, X.sub.87 is G and
X.sub.88 is F, [0205] vii) sequence SEQ ID NO: 203 which
corresponds to sequence SEQ ID NO: 175 in which X.sub.69 to
X.sub.81 are as defined for sequence SEQ ID NO: 176 and X.sub.82 is
P, X.sub.83 is H, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0206] viii) sequence SEQ ID NO:
204 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 176 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is A,
X.sub.87 is G and X.sub.88 is F, [0207] ix) sequence SEQ ID NO: 205
which corresponds to sequence SEQ ID NO: 175 in which X.sub.69 to
X.sub.81 are as defined for sequence SEQ ID NO: 176 and X.sub.82 is
P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is R and X.sub.88 is F, and [0208] x) sequence SEQ ID NO:
210 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 176 and X.sub.82
is P, X.sub.83 is H, X.sub.84 is T, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0209] the peptides having at
least sequence SEQ ID NO: 177 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is L,
X.sub.72 is F, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is K and X.sub.81 is L, and at most a sequence chosen from
the following sequences: [0210] i) sequence SEQ ID NO: 189 which
corresponds to sequence SEQ ID NO: 175 in which X.sub.69 to
X.sub.81 are as defined for sequence SEQ ID NO: 177 and X.sub.82 is
P, X.sub.83 is L, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, and [0211] ii) sequence SEQ ID NO:
190 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.8, are as defined for sequence SEQ ID NO: 177 and X.sub.82
is P, X.sub.83 is P, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0212] the peptides having at
least sequence SEQ ID NO: 178 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is L,
X.sub.72 is F, X.sub.73 is D, X.sub.74est V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is T,
X.sub.80 is K and X.sub.81 is L, and at most sequence SEQ ID NO:
191 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 178 and X.sub.82
is P, X.sub.83 is L, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0213] the peptides having at
least sequence SEQ ID NO: 179 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is Q, X.sub.70 is P, X.sub.71 is L,
X.sub.72 is F, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is T and X.sub.81 is L, and at most sequence SEQ ID NO:
195 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.8, are as defined for sequence SEQ ID NO: 179 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0214] the peptides having at
least sequence SEQ ID NO: 180 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is A, X.sub.71 is L,
X.sub.72 is F, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is T and X.sub.81 is L, and at most a sequence chosen from
the following sequences: [0215] i) sequence SEQ ID NO: 196 which
corresponds to sequence SEQ ID NO: 175 in which X.sub.69 to
X.sub.8, are as defined for sequence SEQ ID NO: 180 and X.sub.82 is
P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, and [0216] ii) sequence SEQ ID NO:
206 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.8, are as defined for sequence SEQ ID NO: 180 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is P, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0217] the peptides having at
least sequence SEQ ID NO: 181 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is L,
X.sub.72 is F, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is I and X.sub.81 is L, and at most sequence SEQ ID NO:
197 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 181 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0218] the peptides having at
least sequence SEQ ID NO: 182 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is L,
X.sub.72 is F, X.sub.73 is E, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is T and X.sub.81 is L, and at most a sequence chosen from
the following sequences: [0219] i) sequence SEQ ID NO: 198 which
corresponds to sequence SEQ ID NO: 175 in which X.sub.69 to
X.sub.81 are as defined for sequence SEQ ID NO: 182 and X.sub.82 is
P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, and [0220] ii) sequence SEQ ID NO:
209 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 182 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is P, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0221] the peptides having at
least sequence SEQ ID NO: 183 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is F,
X.sub.72 is F, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is T and X.sub.81 is L, and at most a sequence chosen from
the following sequences: [0222] i) sequence SEQ ID NO: 199 which
corresponds to sequence SEQ ID NO: 175 in which X.sub.69 to
X.sub.81 are as defined for sequence SEQ ID NO: 183 and X.sub.82 is
P, X.sub.83 is K, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, and [0223] ii) sequence SEQ ID NO:
200 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 183 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0224] the peptides having at
least sequence SEQ ID NO: 184 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is A, X.sub.71 is L,
X.sub.72 is F, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is N, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is T and X.sub.81 is L, and at most sequence SEQ ID NO:
207 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 184 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0225] the peptides having at
least sequence SEQ ID NO: 185 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is L,
X.sub.72 is F, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is N and X.sub.81 is L, and at most sequence SEQ ID NO:
208 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 185 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0226] the peptides having at
least sequence SEQ ID NO: 186 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is L,
X.sub.72 is Y, X.sub.73 is D, X.sub.74 is V, X.sub.75 is M,
X.sub.76 is Y, X.sub.77 is D, X.sub.78 is V, X.sub.79 is S,
X.sub.80 is T and X.sub.81 is L, and at most sequence SEQ ID NO:
211 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 186 and X.sub.82
is P, X.sub.83 is Q, X.sub.84 is A, X.sub.85 is S, X.sub.86 is S,
X.sub.87 is G and X.sub.88 is F, [0227] the peptides having at
least sequence SEQ ID NO: 187 which corresponds to sequence SEQ ID
NO: 174 in which X.sub.69 is R, X.sub.70 is P, X.sub.71 is L,
X.sub.72 is Y, X.sub.73 is D, X.sub.74 is S, X.sub.75 is R,
X.sub.76 is H, X.sub.77 is D, X.sub.78 is I, X.sub.79 is K,
X.sub.80 is K and X.sub.81 is T, and at most sequence SEQ ID NO:
212 which corresponds to sequence SEQ ID NO: 175 in which X.sub.69
to X.sub.81 are as defined for sequence SEQ ID NO: 187 and X.sub.82
is A, X.sub.83 is L, X.sub.84 is A, X.sub.85 is A, X.sub.86 is A,
X.sub.87 is G and X.sub.88 is F.
[0228] Preferably, the D peptide is chosen from the peptides of
sequences SEQ ID NO: 176 to 187, the peptide of sequence SEQ ID NO:
176 being preferred.
[0229] The A, B, C or D peptides as defined by the preceding
sequences 1 to 212 are novel and also constitute a subject of the
invention.
[0230] Thus, according to the invention:
[0231] the A peptide has at least the amino acid sequence SEQ ID
NO: 1 and at most sequence SEQ ID NO: 2; in particular it is chosen
from the peptides: [0232] having at least sequence SEQ ID NO: 3 and
at most sequence SEQ ID NO: 19, [0233] having at least sequence SEQ
ID NO: 4 and at most a sequence chosen from sequences SEQ ID NO:
20, SEQ ID NO: 24, SEQ ID, NO 32 and SEQ ID NO: 34, [0234] having
at least sequence SEQ ID NO: 5 and at most a sequence chosen from
sequences SEQ ID NO: 21, SEQ ID NO: 28 and SEQ ID NO: 36, [0235]
having at least sequence SEQ ID NO: 6 and at most a sequence chosen
from sequences SEQ ID NO: 22, SEQ ID NO: 27 and SEQ ID NO: 41,
[0236] having at least sequence SEQ ID NO: 7 and at most a sequence
chosen from sequences SEQ ID NO: 23 and SEQ ID NO: 37, [0237]
having at least sequence SEQ ID NO: 8 and at most sequence SEQ ID
NO: 25, [0238] having at least sequence SEQ ID NO: 9 and at most
sequence SEQ ID NO: 26, [0239] having at least sequence SEQ ID NO:
10 and at most sequence SEQ ID NO: 29, [0240] having at least
sequence SEQ ID NO: 11 and at most sequence SEQ ID NO: 30, [0241]
having at least sequence SEQ ID NO: 12 and at most sequence SEQ ID
NO: 31, [0242] having at least sequence SEQ ID NO: 13 and at most
sequence SEQ ID NO: 33, [0243] having at least sequence SEQ ID NO:
14 and at most a sequence chosen from sequences SEQ ID NO: 35 and
SEQ ID NO: 39, [0244] having at least sequence SEQ ID NO: 15 and at
most sequence SEQ ID NO: 38, [0245] having at least sequence SEQ ID
NO: 16 and at most a sequence chosen from sequences SEQ ID NO: 40
and SEQ ID NO: 42, [0246] having at least sequence SEQ ID NO: 17
and at most sequence SEQ ID NO: 43, and [0247] having at least
sequence SEQ ID NO: 18 and at most sequence SEQ ID NO: 44, the
peptides of sequence SEQ ID NO: 3 to 18 being preferred and the
peptide of sequence SEQ ID NO: 3 being particularly preferred,
[0248] the B peptide has at least the amino acid sequence SEQ ID
NO: 45 and at most sequence SEQ ID NO: 46; in particular, the B
peptide is chosen from the peptides: [0249] having at least
sequence SEQ ID NO: 47 and at most sequence SEQ ID NO: 81, [0250]
having at least sequence SEQ ID NO: 48 and at most sequence SEQ ID
NO: 82, [0251] having at least sequence SEQ ID NO: 49 and at most
sequence SEQ ID NO: 83, [0252] having at least sequence SEQ ID NO:
50 and at most a sequence chosen from sequences SEQ ID NO: 84, SEQ
ID NO: 90, SEQ ID NO: 105, SEQ ID NO: 107 and SEQ ID NO: 116,
[0253] having at least sequence SEQ ID NO: 51 and at most a
sequence chosen from sequences SEQ ID NO: 85 and SEQ ID NO: 124,
[0254] having at least sequence SEQ ID NO: 52 and at most a
sequence chosen from sequences SEQ ID NO: 86 and SEQ ID NO: 120,
[0255] having at least sequence SEQ ID NO: 53 and at most sequence
SEQ ID NO: 87, [0256] having at least sequence SEQ ID NO: 54 and at
most a sequence chosen from sequences SEQ ID NO: 88 and SEQ ID NO:
117, [0257] having at least sequence SEQ ID NO: 55 and at most
sequence SEQ ID NO: 89, [0258] having at least sequence SEQ ID NO:
56 and at most a sequence chosen from sequences SEQ ID NO: 91 and
SEQ ID NO: 92, [0259] having at least sequence SEQ ID NO: 57 and at
most sequence SEQ ID NO: 93, [0260] having at least sequence SEQ ID
NO: 58 and at most a sequence chosen from sequences SEQ ID NO: 94
and SEQ ID NO: 110, [0261] having at least sequence SEQ ID NO: 59
and at most sequence SEQ ID NO: 95, [0262] having at least sequence
SEQ ID NO: 60 and at most a sequence chosen from sequences SEQ ID
NO: 96 and SEQ ID NO: 115, [0263] having at least sequence SEQ ID
NO: 61 and at most sequence SEQ ID NO: 97, [0264] having at least
sequence SEQ ID NO: 62 and at most a sequence chosen from sequences
SEQ ID NO: 98 and SEQ ID NO: 109, [0265] having at least sequence
SEQ ID NO: 63 and at most sequence SEQ ID NO: 99, [0266] having at
least sequence SEQ ID NO: 64 and at most sequence SEQ ID NO: 100,
[0267] having at least sequence SEQ ID NO: 65 and at most sequence
SEQ ID NO: 101, [0268] having at least sequence SEQ ID NO: 66 and
at most sequence SEQ ID NO: 102, [0269] having at least sequence
SEQ ID NO: 67 and at most sequence SEQ ID NO: 103, [0270] having at
least sequence SEQ ID NO: 68 and at most sequence SEQ ID NO: 104,
[0271] having at least sequence SEQ ID NO: 69 and at most sequence
SEQ ID NO: 106, [0272] having at least sequence SEQ ID NO: 70 and
at most sequence SEQ ID NO: 108, [0273] having at least sequence
SEQ ID NO: 71 and at most sequence SEQ ID NO: 111, [0274] having at
least sequence SEQ ID NO: 72 and at most sequence SEQ ID NO: 112,
[0275] having at least sequence SEQ ID NO: 73 and at most sequence
SEQ ID NO: 113, [0276] having at least sequence SEQ ID NO: 74 and
at most sequence SEQ ID NO: 114, [0277] having at least sequence
SEQ ID NO: 75 and at most a sequence chosen from sequences SEQ ID
NO: 118 and SEQ ID NO: 119 [0278] having at least sequence SEQ ID
NO: 76 and at most sequence SEQ ID NO: 121, [0279] having at least
sequence SEQ ID NO: 77 and at most sequence SEQ ID NO: 122, [0280]
having at least sequence SEQ ID NO: 78 and at most sequence SEQ ID
NO: 123, [0281] having at least sequence SEQ ID NO: 79 and at most
sequence SEQ ID NO: 125, and [0282] having at least sequence SEQ ID
NO: 80 and at most sequence SEQ ID NO: 126, the peptides of
sequences SEQ ID NO: 47 to 80 being preferred and the peptide of
sequence SEQ ID NO: 47 being particularly preferred,
[0283] the C peptide has at least the amino acid sequence SEQ ID
NO: 127 and at most sequence SEQ ID NO: 128; in particular, the C
peptide is chosen from the peptides: [0284] having at least
sequence SEQ ID NO: 129 and at most a sequence chosen from
sequences SEQ ID NO: 147, SEQ ID NO: 153, SEQ ID NO: 162 and SEQ ID
NO: 167, [0285] having at least sequence SEQ ID NO: 130 and at most
a sequence chosen from sequences SEQ ID NO: 148 and SEQ ID NO: 150,
[0286] having at least sequence SEQ ID NO: 131 and at most sequence
SEQ ID NO: 149, [0287] having at least sequence SEQ ID NO: 132 and
at most a sequence chosen from sequences SEQ ID NO: 151, SEQ ID NO:
155, SEQ ID NO: 168 and SEQ ID NO: 171, [0288] having at least
sequence SEQ ID NO: 133 and at most sequence SEQ ID NO: 152, [0289]
having at least sequence SEQ ID NO: 134 and at most a sequence
chosen from sequences SEQ ID NO: 154 and SEQ ID NO: 169, [0290]
having at least sequence SEQ ID NO: 135 and at most sequence SEQ ID
NO: 156, [0291] having at least sequence SEQ ID NO: 136 and at most
a sequence chosen from sequences SEQ ID NO: 157 and SEQ ID NO: 170,
[0292] having at least sequence SEQ ID NO: 137 and at most sequence
SEQ ID NO: 158, [0293] having at least sequence SEQ ID NO: 138 and
at most sequence SEQ ID NO: 159, [0294] having at least sequence
SEQ ID NO: 139 and at most sequence SEQ ID NO: 160, [0295] having
at least sequence SEQ ID NO: 140 and at most sequence SEQ ID NO:
161, [0296] having at least sequence SEQ ID NO: 141 and at most
sequence SEQ ID NO: 163, [0297] having at least sequence SEQ ID NO:
142 and at most sequence SEQ ID NO: 164, [0298] having at least
sequence SEQ ID NO: 143 and at most sequence SEQ ID NO: 165, [0299]
having at least sequence SEQ ID NO: 144 and at most sequence SEQ ID
NO: 166, [0300] having at least sequence SEQ ID NO: 145 and at most
sequence SEQ ID NO: 172, and [0301] having at least sequence SEQ ID
NO: 146 and at most sequence SEQ ID NO: 173, the peptides of
sequence SEQ ID NO: 129 to 146 being preferred and the peptide of
sequence SEQ ID NO: 129 being particularly preferred, and
[0302] the D peptide has at least the amino acid sequence SEQ ID
NO: 174 and at most sequence SEQ ID NO: 175; in particular, the D
peptide is chosen from the peptides: [0303] having at least
sequence SEQ ID NO: 176 and at most a sequence chosen from
sequences SEQ ID NO: 188, SEQ ID NO: 192, SEQ ID NO: 193, SEQ ID
NO: 194, SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO:
204, SEQ ID NO: 205 and SEQ ID NO: 210, [0304] having at least
sequence SEQ ID NO: 177 and at most a sequence chosen from
sequences SEQ ID NO: 189 and SEQ ID NO: 190, [0305] having at least
sequence SEQ ID NO: 178 and at most sequence SEQ ID NO: 191, [0306]
having at least sequence SEQ ID NO: 179 and at most sequence SEQ ID
NO: 195, [0307] having at least sequence SEQ ID NO: 180 and at most
a sequence chosen from sequences SEQ ID NO: 196 and SEQ ID NO: 206,
[0308] having at least sequence SEQ ID NO: 181 and at most sequence
SEQ ID NO: 197, [0309] having at least sequence SEQ ID NO: 182 and
at most a sequence chosen from sequences SEQ ID NO: 198 and SEQ ID
NO: 209, [0310] having at least sequence SEQ ID NO: 183 and at most
a sequence chosen from sequences SEQ ID NO: 199 and SEQ ID NO: 200,
[0311] having at least sequence SEQ ID NO: 184 and at most sequence
SEQ ID NO: 207, [0312] having at least sequence SEQ ID NO: 185 and
at most sequence SEQ ID NO: 208, [0313] having at least sequence
SEQ ID NO: 186 and at most sequence SEQ ID NO: 211, and [0314]
having at least sequence SEQ ID NO: 187 and at most sequence SEQ ID
NO: 212,
[0315] the peptides of sequence SEQ ID NO: 176 to 187 being
preferred and the peptide of sequence SEQ ID NO: 176 being
particularly preferred.
[0316] The expression "the A peptide has at least the amino acid
sequence SEQ ID NO: 1 and at most sequence SEQ ID NO: 2" signifies
that the N-terminal end is delimited by the amino acid situated at
one of positions 1 to 8 of SEQ ID NO: 2 and the C-terminal end is
delimited by the amino acid situated at one of positions 38 to 46
of SEQ ID NO: 2.
[0317] Thus the A peptide has at least the 31 consecutive amino
acids of sequence SEQ ID NO: 1 and at most the 46 consecutive amino
acids of sequence SEQ ID NO: 2.
[0318] Sequence SEQ ID NO: 2 includes the 31 amino acids of
sequence SEQ ID NO: 1.
[0319] Thus the A peptide of the invention always has at least the
31 amino acids of sequence SEQ ID NO: 1 and at most 15 additional
amino acids distributed on both sides of these 31 amino acids
within the limit of sequence SEQ ID NO: 2. For example, an A
peptide of the invention can comprise 33 amino acids constituted by
31 amino acids of SEQ ID NO: 1 and either 2 N-terminal amino acids,
or 2 C-terminal amino acids, or 1 N-terminal amino acid and a
C-terminal amino acid.
[0320] The expression "the B peptide has at least the amino acid
sequence SEQ ID NO: 45 and at most sequence SEQ ID NO: 46"
signifies that the N-terminal end is delimited by the amino acid
situated at one of positions 1 to 12 of SEQ ID NO: 46 and the
C-terminal end is delimited by the amino acid situated at one of
positions 56 to 63 of SEQ ID NO: 46.
[0321] Thus the B peptide has at least the 45 consecutive amino
acids of sequence SEQ ID NO: 45 and at most the 63 consecutive
amino acids of sequence SEQ ID NO: 46.
[0322] Sequence SEQ ID NO: 46 includes the 45 amino acids of
sequence SEQ ID NO: 45.
[0323] Thus the B peptide of the invention always has at least the
45 amino acids of sequence SEQ ID NO: 45 and at most 18 additional
amino acids distributed on both sides of these 45 amino acids
within the limit of sequence SEQ ID NO: 46.
[0324] The expression "the C peptide has at least the amino acid
sequence SEQ ID NO: 127 and at most sequence SEQ ID NO: 128"
signifies that the N-terminal end is delimited by the amino acid
situated at one of positions 1 to 23 of SEQ ID NO: 128 and the
C-terminal end is delimited by the amino acid situated at one of
positions 54 to 57 of SEQ ID NO: 128.
[0325] Thus the C peptide has at least the 32 consecutive amino
acids of sequence SEQ ID NO: 127 and at most the 57 consecutive
amino acids of sequence SEQ ID NO: 128.
[0326] Sequence SEQ ID NO: 128 includes the 32 amino acids of
sequence SEQ ID NO: 127.
[0327] Thus the C peptide of the invention always has at least the
32 amino acids of sequence SEQ ID NO: 127 and at most 25 additional
amino acids distributed on both sides of these 32 amino acids
within the limit of sequence SEQ ID NO: 128.
[0328] The expression "the D peptide has at least the amino acid
sequence SEQ ID NO: 174 and at most sequence SEQ ID NO: 175"
signifies that the N-terminal end is delimited by the amino acid
situated at one of positions 1 to 4 of SEQ ID NO: 175 and the
C-terminal end is delimited by the amino acid situated at one of
positions 32 to 44 of SEQ ID NO: 175.
[0329] Thus the D peptide has at least the 29 consecutive amino
acids of sequence SEQ ID NO: 174 and at most the 44 consecutive
amino acids of sequence SEQ ID NO: 175.
[0330] Sequence SEQ ID NO: 175 includes the 29 amino acids of
sequence SEQ ID NO: 174.
[0331] Thus the D peptide of the invention always has at least the
29 amino acids of sequence SEQ ID NO: 174 and at most 15 additional
amino acids distributed on both sides of these 29 amino acids
within the limit of sequence SEQ ID NO: 175.
[0332] The B and C peptides contain novel epitopes having a strong
immunogenic power.
[0333] Another subject of the invention relates to the B' epitope,
contained in the B peptide and situated between positions 1038 and
1047 of the viral polyprotein, which possesses the following amino
acid sequence SEQ ID NO: 213: [0334]
GX.sub.18X.sub.19X.sub.20X.sub.21X.sub.22X.sub.23TSL
[0335] in which X.sub.18 is L or V, X.sub.19 is L or F, X.sub.20 is
G or S, X.sub.21 is C or T, X.sub.22 is I or V and X.sub.23 is I or
V.
[0336] According to one embodiment of the invention, the B' epitope
is chosen from the following epitopes: [0337] the epitope of
sequence SEQ ID NO: 214 which corresponds to sequence SEQ ID NO:
213 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G, X.sub.21
is C, X.sub.22 is I and X.sub.23 is I, [0338] the epitope of
sequence SEQ ID NO: 215 which corresponds to sequence SEQ ID NO:
213 in which X.sub.18 is L, X.sub.19 is F, X.sub.20 is G, X.sub.21
is C, X.sub.22 is I and X.sub.23 is I, [0339] the epitope of
sequence SEQ ID NO: 216 which corresponds to sequence SEQ ID NO:
213 in which X.sub.18 is V, X.sub.19 is L, X.sub.20 is G, X.sub.21
is C, X.sub.22 is V and X.sub.23 is I, [0340] the epitope of
sequence SEQ ID NO: 217 which corresponds to sequence SEQ ID NO:
213 in which X.sub.18 is L, X.sub.19 is V, X.sub.20 is G, X.sub.21
is C, X.sub.22 is I and X.sub.23 is I, [0341] the epitope of
sequence SEQ ID NO: 218 which corresponds to sequence SEQ ID NO:
213 in which X.sub.18 is L, X.sub.19 is L, X.sub.20 is G, X.sub.21
is C, X.sub.22 is I and X.sub.23 is V, [0342] the epitope of
sequence SEQ ID NO: 219 which corresponds to sequence SEQ ID NO:
213 in which X.sub.18 is L, X.sub.19 is F, X.sub.20 is G, X.sub.21
is C, X.sub.22 is I and X.sub.23 is V, and [0343] the epitope of
sequence SEQ ID NO: 220 which corresponds to sequence SEQ ID NO:
213 in which X.sub.18 is L, X.sub.19 is F, X.sub.20 is S, X.sub.21
is T, X.sub.22 is I and X.sub.23 is I.
[0344] Preferably, the B' epitope possesses at least one of the
following characteristics: [0345] it has sequence SEQ ID NO: 214
and [0346] it is restricted to HLA-A2.
[0347] Another subject of the invention relates to the C' epitope,
contained in the C peptide and situated between positions 1789 and
1821 of the viral polyprotein, which possesses the following amino
acid sequence SEQ ID NO: 221: [0348]
SPLX.sub.52X.sub.53X.sub.54X.sub.55TL
[0349] in which X.sub.52 is T, S or A, X.sub.53 is T or I, X.sub.54
is Q, S or G and X.sub.55 is N, Q, H, S, Y or T.
[0350] According to one embodiment of the invention, the C' epitope
is chosen from the following epitopes: [0351] the epitope of
sequence SEQ ID NO: 222 which corresponds to sequence SEQ ID NO:
221 in which X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q and
X.sub.55 is N, [0352] the epitope of sequence SEQ ID NO: 223 which
corresponds to sequence SEQ ID NO: 221 in which X.sub.52 is T,
X.sub.53 is T, X.sub.54 is S and X.sub.55 is Q, [0353] the epitope
of sequence SEQ ID NO: 224 which corresponds to sequence SEQ ID NO:
221 in which X.sub.52 is T, X.sub.53 is T, X.sub.54 is G and
X.sub.55 is Q, [0354] the epitope of sequence SEQ ID NO: 225 which
corresponds to sequence SEQ ID NO: 221 in which X.sub.52 is T,
X.sub.53 is T, X.sub.54 is Q and X.sub.55 is H, [0355] the epitope
of sequence SEQ ID NO: 226 which corresponds to sequence SEQ ID NO:
221 in which X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q and
X.sub.55 is S, [0356] the epitope of sequence SEQ ID NO: 227 which
corresponds to sequence SEQ ID NO: 221 in which X.sub.52 is T,
X.sub.53 is T, X.sub.54 is Q and X.sub.55 is Y, [0357] the epitope
of sequence SEQ ID NO: 228 which corresponds to sequence SEQ ID NO:
221 in which X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q and
X.sub.55 is T, [0358] the epitope of sequence SEQ ID NO: 229 which
corresponds to sequence SEQ ID NO: 221 in which X.sub.52 is T,
X.sub.53 is I, X.sub.54 is Q and X.sub.55 is H, [0359] the epitope
of sequence SEQ ID NO: 230 which corresponds to sequence SEQ ID NO:
221 in which X.sub.52 is S, X.sub.53 is T, X.sub.54 is Q and
X.sub.55 is N, [0360] the epitope of sequence SEQ ID NO: 231 which
corresponds to sequence SEQ ID NO: 221 in which X.sub.52 is A,
X.sub.53 is T, X.sub.54 is Q and X.sub.55 is Y, and [0361] the
epitope of sequence SEQ ID NO: 232 which corresponds to sequence
SEQ ID NO: 221 in which X.sub.52 is T, X.sub.53 is T, X.sub.54 is Q
and X.sub.55 is Q.
[0362] Preferably, the C' epitope possesses at least one of the
following characteristics: [0363] it has sequence SEQ ID NO: 222
and [0364] it is restricted to HLA-B7.
[0365] The compositions of the invention, apart from the A to D
peptides, can also contain the B' and C' epitopes, which
constitutes another subject of the invention.
[0366] In a preferred embodiment, the peptide composition of the
present invention comprises at least two peptides as defined above
which are fused each other.
[0367] The present invention also relates to a novel fusion peptide
comprising at least two peptides as defined above which are fused
each other. The term "fusion" as used herein means that the amino
acids of the first peptide and of the second peptide are fused in
one polypeptide chain, preferably by in frame fusion of the
corresponding coding nucleotide sequences. Advantageously, the
fusion peptide of the invention comprises a fusion of at least two
peptides chosen from the group consisting of A, B, C, D, B' and C'
peptides as defined above, as well as pharmaceutical compositions
containing such a fusion peptide and its use in particular as a
vaccine, for the preparation of a medicament intended for the
inhibition or prevention of an infection caused by the hepatitis C
virus, and as a diagnostic composition.
[0368] A preferred fusion peptide comprises a fusion of at least
four peptides chosen from the group consisting of A, B, C, and D
peptides. A more preferred peptide composition comprises a fusion
of A to D peptides with B peptide located at the N-terminus which
is fused to A peptide which is fused to C peptide which is fused to
D peptide (B/A/C/D fusion peptide). In particular, the A peptide
comprised in the fusion peptide of the invention has at least SEQ
ID NO: 3 and at most SEQ ID NO: 19. Independently or in combination
with the other peptides, the B peptide comprised in the fusion
peptide of the invention has at least SEQ ID NO: 47 and at most SEQ
ID NO: 81. Independently or in combination with the other peptides,
the C peptide comprised in the fusion peptide of the invention has
at least SEQ ID NO: 129 and at most SEQ ID NO: 147, SEQ ID NO: 153,
SEQ ID NO: 162 or SEQ ID NO: 167. Independently or in combination
with the other peptides, the D peptide comprised in the fusion
peptide of the invention has at least SEQ ID NO: 176 and at most
SEQ ID NO: 188, SEQ ID NO: 192, SEQ ID NO: 193 or SEQ ID NO: 194,
SEQ ID NO: 201, SEQ ID NO: 202, SEQ ID NO: 203, SEQ ID NO: 204, SEQ
ID NO: 205 or SEQ ID NO: 210. In a preferred embodiment, the A
peptide comprised in the fusion peptide of the invention has the
amino acid sequence defined in SEQ ID NO: 3, the B peptide
comprised in the fusion peptide of the invention has the amino acid
sequence defined in SEQ ID NO: 47, the C peptide comprised in the
fusion peptide of the invention has the amino acid sequence defined
in SEQ ID NO: 129 and the D peptide comprised in the fusion peptide
of the invention has the amino acid sequence defined in SEQ ID NO:
176. Even more preferably, the fusion peptide of the invention
comprises the amino acid sequence defined in SEQ ID NO: 234.
[0369] The present invention also relates to the nucleotide
sequences coding for any one of the A to D peptides as defined by
sequences SEQ ID NO: 1 to 212 and B' and C' epitopes as defined by
sequences SEQ ID NO: 213 to 232 and any one of the fusion peptides
as defined above, with a special preference for the fusion peptide
comprises the amino acid sequence defined by SEQ ID NO: 234.
[0370] The peptides of the invention can be obtained by the genetic
engineering technique which comprises the stages of: [0371] culture
of a microorganism or of eukaryotic cells transformed using a
nucleotide sequence according to the invention and [0372] recovery
of the peptide produced by said microorganism or said eukaryotic
cells.
[0373] This technique is well known to a person skilled in the art.
For more details concerning this, reference can be made to the
following work: Recombinant DNA Technology I, Editors Ales Prokop,
Raskesh K Bajpai; Annals of the New York Academy of Sciences,
Volume 646, 1991.
[0374] The peptides of the invention can also be prepared by the
standard peptide syntheses well known to a person skilled in the
art.
[0375] The nucleotide sequences according to the invention can be
prepared by chemical synthesis and genetic engineering using the
techniques well known to a person skilled in the art and described
for example in Sambrook J. et al., Molecular Cloning: A Laboratory
Manual, 1989.
[0376] The nucleotide sequences of the invention can be inserted
into expression vectors in order to obtain the compositions or
peptides of the invention.
[0377] Thus, another subject of the invention is the expression
vectors comprising a nucleotide sequence of the invention, as well
as the means necessary for its expression. Such means necessary for
expression are well known in the art and can vary according to the
host cell, the expression vector and the level of expression
desired.
[0378] As expression vectors, there can be mentioned for example
the plasmids, the viral vectors of the vaccine virus type,
adenovirus, baculovirus, poxvirus, bacterial vectors of salmonella
type, BCG. Such vectors and methods of using and making them are
well known in the art (see for example <<Nonviral Vectors for
gene Therapy>>, 2001, edited by M. Findeis, Humana Press;
<<Adenoviral Vectors for Gene Therapy>>, 2002, edited
by Curiel and Douglas, Elsevier Science, Academic Press; and
<<Vaccinia Virus and poxyirology>>, 2004, edited by S.
Isaacs, Humana Press). The term "viral vector" as used herein
encompasses vector DNA as well as viral particles generated thereof
by conventional technologies.
[0379] In one embodiment, the vector of the invention is an
adenoviral vector. It can be derived from a variety of human or
animal sources. Any serotype can be employed from the adenovirus
serotypes 1 through 51, with a special preference for human
adenoviruses 2 (Ad2), 5 (Ad5), 6 (Ad6), 11 (Ad11), 24 (Ad24) and 35
(Ad35). The cited adenoviruses are available from the American Type
Culture Collection (ATCC, Rockville, Md.), and have been the
subject of numerous publications describing their sequence,
organization and methods of producing, allowing the artisan to
apply them (see for example U.S. Pat. No. 6,133,028; U.S. Pat. No.
6,110,735; WO02/40665; WO00/50573; EP 1 016 711; Vogels et al.,
2003, J. Virol. 77: 8263-8271). Preferably, the adenoviral vector
of the invention is replication-defective (see for example
WO94/28152; Lusky et al., 1998, J. Virol 72, 2022-2032). Preferred
replication-defective adenoviral vectors are E1-defective with an
E1 deletion extending from approximately positions 459 to 3328 or
from approximately positions 459 to 3510 (by reference to the
sequence of the human adenovirus type 5 disclosed in the GeneBank
under the accession number M 73260 and in Chroboczek et al., 1992,
Virol. 186, 280-285). The cloning capacity can further be improved
by deleting additional portion(s) of the adenoviral genome (all or
part of the non essential E3 region or of other essential E2, E4
regions). The nucleotide sequence of the present invention can be
inserted in any location of the adenoviral genome. Preferably, it
is inserted in replacement of the E1 region. It may be positioned
in sense or antisense orientation relative to the natural
transcriptional direction of the region in question.
[0380] In another embodiment the vector of the invention is derived
from a poxvirus. It may be obtained from any member of the
poxyiridae, in particular canarypox, fowlpox and vaccinia virus,
the latter being preferred. Suitable vaccinia viruses include
without limitation the Copenhagen strain (Goebel et al., 1990,
Virol. 179: 247-266 and 517-563; Johnson et al., 1993, Virol. 196:
381-401), the Wyeth strain and the modified Ankara (MVA) strain
(Antoine et al., 1998, Virol. 244: 365-396). The general conditions
for constructing recombinant poxvirus are well known in the art
(see for example EP 206 920; Mayr et al., 1975, Infection 3: 6-14;
Sutter and Moss, 1992, Proc. Natl. Acad. Sci. USA 89: 10847-10851;
U.S. Pat. No. 6,440,422). The nucleotide sequence of the present
invention is preferably inserted within the poxyiral genome in a
non-essential locus. Thymidine kinase gene is particularly
appropriate for insertion in Copenhagen vaccinia vectors (Hruby et
al., 1983, Proc. Natl. Acad. Sci USA 80: 3411-3415; Weir et al.,
1983, J. Virol. 46: 530-537) and deletion II or III for insertion
in MVA vector (Meyer et al., 1991, J. Gen. Virol. 72: 1031-1038;
Sutter et al., 1994, Vaccine 12: 1032-1040).
[0381] In another embodiment, the expression vector of the
invention can be optionally coupled or complexed to conventional
drug delivery systems (e.g. lipid or polymer-based liposomes,
nanoparticles, etc. such as those described for example in Mahato
et al., 1998, Human Gene Ther. 9: 2083-2099 and Allen et al., 2004,
Science 303: 18181822).
[0382] By "means necessary for the expression" it is meant any
means which make it possible to obtain the peptide or fusion
peptide of the invention, such as in particular, a promoter, a
transcription terminator, a replication origin and preferably a
selection marker.
[0383] The promoter used in the context of the invention can be of
any origin, e.g. viral, cellular or synthetic and be ubiquitous
providing constitutive expression or regulable providing for
example specific expression in a particular cell type or under
specific conditions. It can further be operably linked to an
enhancer. Suitable viral promoters include without limitation early
promoters obtained from RSV (Rous Sarcoma Virus), SV40 (Simian
Virus), and CMV (Cytomegalovirus; Boshart et al., 1985, Cell 41,
521-530), as well as the TK (Thymidine kinase) promoter of HSV-1
virus (Herpes Virus Simplex-1), the major late adenovirus promoter
(MLP) and vaccinia promoters (e.g. 7.5K, H5R, TK, p28, p11 and K1L
promoters). One may use synthetic promoters such as those described
in particular by Chakrabarti et al. (1997, Biotechniques 23:
1094-1097), Hammond et al. (1997, J. Virological Methods 66:
135-138). Suitable cellular promoters include any promoter driving
expression of cellular genes with a special interest for liver
specific promoters such as those of phosphoglycero kinase (PGK;
Adra et al., 1987, Gene 60: 65-74), albumin (Pinkert et al., 1987,
Genes Dev. 1: 268-277), phosphoenol pyruvate carboxy kinase (PEPCK)
(Eisenberger et al., 1992, Mol. Cell Biol. 12: 1396-1403),
cholesterol 7-alpha hydroylase (CYP-7) (Lee et al., 1994, J. Biol.
Chem. 269: 14681-14689), alpha-1 antitrypsin (Ciliberto et al.,
1985, Cell 41: 531-540), transferrine (Mendelzon et al., 1990,
Nucleic Acids Res. 18: 5717-5721); and facteur IX (U.S. Pat. No.
5,814,716) genes.
[0384] The vectors of the invention may also comprise one or more
additional means in order to improve the transcription rate or
level of the nucleotide sequence of the invention in a given host
cell, its stability, nuclear RNA transport and/or translation rate
or level of the mRNA. Such means are well known by the skilled
person and include for example 5', 3' non-coding sequences,
intervening sequences, splicing sequences, Shine-Dalgarno sequence,
Kozak sequence and initiator methionine.
[0385] The vectors of the invention can also comprise sequences
necessary for the screening of the peptides towards particular cell
compartments. An example of screening can be screening towards the
endoplasmic reticulum obtained using orientation sequences of the
type of the leader sequence originating from the protein E3 of the
adenovirus (Ciernik I. F., et al., The Journal of Immunology, 1999,
162: 3915-3925).
[0386] The expression vectors of the invention can comprise either
a single nucleotide sequence coding for any one of the peptides of
the invention, or at least two nucleotide sequences, it being
understood that each nucleotide sequence codes for a peptide of
different type.
[0387] According to one embodiment of the invention, the expression
vectors include two nucleotide sequences coding for the A and B, A
and C, A and D, B and C, B and D, or D and C peptides.
[0388] Preferably, the expression vectors include two nucleotide
sequences coding for the A and B, A and C, B and D, or C and D
peptides, the vectors comprising two sequences coding for the A and
B, C and D, and B and D peptides being particularly preferred.
[0389] According to another embodiment, the expression vectors
include three nucleotide sequences coding for the A, B and C, A, B
and D, A, C and D, or B, C and D peptides.
[0390] According to yet another method, the expression vectors
include four nucleotide sequences coding for the A to D
peptides.
[0391] Preferably, the vectors include three nucleotide sequences
coding for the A, B and C, A, C and D, and B, C and D peptides.
More preferably, the vectors include four nucleotide sequences
coding for the A, B, C, and D peptides. In this case, the order of
the nucleotide sequences is relatively unimportant, as in the
preceding combinations, such that the following combinations, given
relative to the peptides, are comprised within the scope of the
invention: A/B/C/D, A/B/D/C, A/C/B/D, A/C/D/B, A/D/B/C, A/D/C/B,
B/A/C/D, B/A/D/C, B/C/A/D, B/C/D/A, B/D/A/C, B/D/C/A, C/A/B/D,
C/A/D/B, C/B/A/D, C/B/D/A, C/D/A/B, C/D/B/A, D/A/B/C, D/A/C/B,
D/B/A/C, D/B/C/A, D/C/A/B and D/C/B/A. The more preferred
combination is B/A/C/D, and especially the B/A/C/D fusion peptide
previously defined.
[0392] The expression vectors of the invention can also comprise at
least one nucleotide sequence coding for any one of the B' and C'
epitopes as defined previously, which constitutes another
embodiment of the invention.
[0393] Thus, the vectors of the invention can comprise for example:
[0394] a nucleotide sequence coding for one of the following
epitopes: B' and C', [0395] two nucleotide sequences coding for the
B' and C' epitopes or [0396] several repetitive sequences coding
for the B' epitope, the C' epitope or both.
[0397] The vectors of the invention can also comprise from two to
four nucleotide sequences chosen from the sequences coding for the
A, B, C, D peptides and the B' and C' epitopes, it being understood
that: [0398] the nucleotide sequence coding for the B peptide and
the nucleotide sequence coding for the B' epitope are not present
at the same time, and [0399] the nucleotide sequence coding for the
C peptide and the nucleotide sequence coding for the C' epitope are
not present at the same time.
[0400] Thus, for example, the vectors of the invention can comprise
the following combinations, given relative to said peptides and
epitopes: A/B', A/C', A'/D, B/C', B'/C, B'/D, C'/D, A/B'/C,
A/B'/C', A/B/C', A/B'/D, A/C'/D, B'/C/D, B/C'/D, B'/C'/D, A/B'/C/D,
A/B/C'/D and A/B'/C'/D.
[0401] Of course, as previously, the order of the nucleotide
sequences in the expression vectors is relatively unimportant.
[0402] When the expression vectors of the invention include several
nucleotide sequences, said sequences can be linked to each other
directly, or via spacing or binding agents which are typically made
up of small neutral molecules such as amino acids or amino acid
mimetics which typically have a neutral charge under physiological
conditions. Direct linkage of the peptide-encoding nucleotide
sequences each other in a fusion peptide is preferred in the
context of the invention.
[0403] In a preferred embodiment of the invention, the at least two
nucleotide sequences are linked in the same reading frame. The
expression "linked in the same reading frame" implies that in said
reading frame, there is no stop codon between said two nucleotide
sequences.
[0404] As spacing agents, there can be mentioned the Ala, Gly
residues or other neutral spacing agents of non-polar amino acids
or neutral polar amino acids.
[0405] These amino acid spacers have at least one or two residues
and usually from 3 to 6 residues.
[0406] A subject of the invention is also the microorganisms and
eukaryotic cells transformed by an expression vector of the
invention. In the context of the invention, the term
"transformation" or "transformed" has to be understood as meaning
"introduction" or "introduced" in a host cell. Any routine method
can be used in the art to "transform" a nucleotide sequence or a
vector in a host cell, e.g. a microorganism or eukaryotic cell.
Such methods include, but are not limited to, microinjection
(Capechi et al., 1980, Cell 22, 479-488), CaPO.sub.4-- mediated
transfection (Chen and Okayama, 1987, Mol. Cell Biol. 7,
2745-2752), DEAE-dextran-mediated transfection, electroporation
(Chu et al., 1987, Nucleic Acid Res. 15: 1311-1326),
lipofection/liposome fusion (Felgner et al., 1987, Proc. Natl.
Acad. Sci. USA 84: 7413-7417), particle bombardement (Yang et al.,
1990, Proc. Natl. Acad. Sci. USA 87: 9568-9572), gene guns,
transduction as well as viral infection. For example, in the
context of the invention, a viral vector can be transformed in the
host cell by transfection of its genome or by infection of a viral
particle. The term "host cell" should be understood broadly without
any limitation concerning microorganisms and eukaryotic cells
including isolated cells or cells organized in particular
structures such as tissues and organs. The host cells may be of a
unique type of cells or a group of different types of cells and
encompass cultured cell lines, primary cells and proliferative
cells.
[0407] When a composition of the invention containing at least two
A to D peptides of the invention is to be obtained, the
microorganisms or eukaryotic cells are transformed by an expression
vector containing at least two nucleotide sequences, or they are
cotransformed by at least two expression vectors containing a
single nucleotide sequence, each vector coding for a peptide of
different type.
[0408] According to one embodiment of the invention, the
microorganisms and eukaryotic cells are cotransformed with: [0409]
two vectors coding respectively for the A and B, A and C, A and D,
B and C, B and D, or C and D peptides, [0410] three vectors coding
respectively for the A, B and C, A, B and D, A, C and D, or B, C
and D peptides, or [0411] four vectors coding respectively for the
A, B, C and D peptides.
[0412] Similarly, when a composition of the invention is to be
obtained containing at least two B' and C' epitopes, or at least
one B' or C' epitope and at least one A to D peptide, the
microorganisms or eukaryotic cells are transformed by a single
vector coding for the desired combination of epitopes or
epitopes/peptides or by several vectors each coding for each
constituent of the desired combination.
[0413] As examples of microorganisms which are appropriate for the
purposes of the invention, there can be mentioned yeasts, such as
those of the following families: Saccharomyces,
Schizosaccharomyces, Kluveromyces, Pichia, Hanseluna, Yarowia,
Schwaniomyces, Zygosaccharomyces, Saccharomyces cerevisiae,
Saccharomyces carlsbergensis and Kluveromyces lactis being
preferred; and bacteria, such as E. coli and those of the following
families: Lactobacillus, Lactococcus, Salmonella, Streptococcus,
Bacillus and Streptomyces.
[0414] As examples of eukaryotic cells, there can be mentioned
cells originating from animals such as mammals, reptiles, insects
and equivalent. The preferred eukaryotic cells are cells
originating from the Chinese hamster (CHO cells), monkey (COS and
Vero cells), baby hamster kidney (BHK cells), pig kidney (PK 15
cells) and rabbit kidney (RK13 cells, human osteosarcoma cell lines
(143 B cells), human HeLa cell lines and human hepatoma cell lines
(Hep G2 cell type), as well as insect cell lines (for example of
Spodoptera frugiperda).
[0415] The host cells can be supplied in cultures in suspension or
in vials, in tissue cultures, organ cultures and equivalent. The
host cells can also be from transgenic animals. Host cells of the
present invention can be cultured in conventional fermentation
bioreactors, flasks, and petri plates. Culturing can be carried out
at a temperature, pH and oxygen content appropriate for a given
host cell. No attempts to describe in detail the various methods
known for the production of polypeptides in microorganisms and
eukaryote cells will be made here.
[0416] The peptides and fusion peptides of the invention can be
purified from the producing host cells by well-known purification
methods including ammonium sulfate precipitation, acid extraction,
gel electrophoresis, filtration and chromatographic methods (e.g.
reverse phase, size exclusion, ion exchange, affinity,
phosphocellulose, hydrophobic-interaction, hydroxylapatite, or high
performance liquid chromatography). The conditions and technology
used to purify a particular peptide or fusion peptide of the
invention will depend on factors such as net charge, molecular
weight, hydrophobicity, hydrophilicity and will be apparent to
those having skill in the art. Moreover, the level of purification
will depend on the intended use.
[0417] The invention also relates to antibodies directed against
one of the abovementioned peptides of the invention or against one
of the peptide compositions of the invention as defined previously,
or also against one of the epitopes of the invention.
[0418] The antibodies according to the invention are either
polyclonal or monoclonal antibodies.
[0419] The abovementioned polyclonal antibodies can be obtained by
immunization of an animal with at least one viral antigen of
interest, followed by the recovery of the sought antibodies in
purified form, by taking a sample of the serum of said animal, and
separation of said antibodies from the other constituents of the
serum, in particular by affinity chromatography on a column on
which an antigen specifically recognized by the antibodies, in
particular a viral antigen of interest, is fixed.
[0420] The monoclonal antibodies can be obtained by the hybridoma
technique, the general principle of which is recalled
hereafter.
[0421] Firstly, an animal, generally a mouse, (or cells in culture
within the framework of in vitro immunizations) is immunized with a
viral antigen of interest, the B lymphocytes of which are then
capable of producing antibodies against said antigen. These
antibody-producing lymphocytes are then fused with "immortal"
myelomatous cells (murine in the example) in order to produce
hybridomas. From the heterogeneous mixture of the cells thus
obtained, a selection is then made of cells capable of producing a
particular antibody and multiplying indefinitely. Each hybridoma is
multiplied in clone form, each leading to the production of a
monoclonal antibody the recognition properties of which vis-a-vis
the viral antigen of interest can be tested for example in ELISA,
by immunotransfer in one or two dimensions, in immunofluorescence,
or using a biocaptor. The monoclonal antibodies thus selected are
subsequently purified in particular according to the affinity
chromatography technique described above.
[0422] The compositions of the invention, containing at least two A
to D peptides or at least one of the B' and C' epitopes or a fusion
peptide as described previously, are particularly effective for the
inhibition, prevention and treatment of the virus or infection of
patients carrying the virus belonging more particularly to the
genotypes 1a, 1b and 4, in such a manner that its use for the
preparation of a medicament constitutes another subject of the
invention.
[0423] The present invention also relates to a pharmaceutical
composition, in particular a vaccine, containing as active
ingredient at least two different peptides chosen from the A to D
peptides as defined previously, or at least two nucleotide
sequences as described previously, placed under the control of
elements necessary for a constitutive and/or inducible expression
of said peptides, or at least one of the antibodies as defined
previously, or also at least one of the B' and C' epitopes as
defined previously, or at least one of their nucleotide sequences,
in combination with a pharmaceutically appropriate vehicle.
[0424] The present invention also relates to a pharmaceutical
composition, in particular vaccine, containing as active ingredient
at least a fusion peptide as defined previously or at least a
nucleotide sequence or a vector or a host cell as defined
previously encoding the fusion peptide of the invention, placed
under the control of the elements necessary for a constitutive
and/or inducible expression of said fusion peptide in a given host
cell, in combination with a pharmaceutically appropriate
vehicle.
[0425] By elements necessary for a constitutive expression of the
peptides/fusion peptide, is meant a ubiquitous or specific promoter
of the eukaryotic cells as previously described.
[0426] As elements necessary for an inducible expression of the
peptides, there can be mentioned the elements of regulation of the
operon of E. coli for resistance to tetracycline (Gossen M. and al,
Proc Natl Acad Sci USA, 89: 5547-5551 (1992).
[0427] Of course, a person skilled in the art will easily determine
the pharmaceutically appropriate vehicle and the quantity of active
ingredient to be used as a function of the constituents of the
pharmaceutical composition. Preferably, the composition of the
invention contains the active ingredient or a combination of active
ingredients at a dose sufficient for the alleviation of one or more
symptoms normally associated with the disease or condition desired
to be treated. When prophylactic use is concerned, this means a
dose sufficient to prevent or to delay the establishment of a
chronic HCV infection. For example, the active ingredient or a
combination thereof is preferably contained in the composition of
the invention at a dose sufficient for inducing a specific immune
response in the treated host (e.g. resulting in the development of
an anti-HCV response, whether humoral or cellular or both, with a
special preference for a cytolytic T cell response).
[0428] The appropriate dosage can be adapted as a function of
various parameters, in particular the mode of administration; the
composition employed; the age, health, and weight of the patient to
be treated; the nature and extent of symptoms; kind of concurrent
treatment; the frequency of treatment; and/or the need for
prevention or therapy. Further refinement of the calculations
necessary to determine the appropriate dosage for treatment is
routinely made by a practitioner, in the light of the relevant
circumstances. For general guidance, suitable dosage for a
virus-comprising composition (e.g. vaccinia or adenovirus
particles) varies from about 10.sup.5 to 10.sup.13 iu (infectious
units), desirably from about 10.sup.6 and 10.sup.12 iu. A
composition based on vector plasmids may be administered in doses
of between 10 .mu.g and 20 mg, advantageously between 100 .mu.g and
2 mg. A peptide composition may be administered in one or more
doses of between 10 ng and 20 mg, with a special preference for a
dosage from about 0.1 .mu.g to about 2 mg of peptide(s) per kg body
weight. The administration may take place in a single dose or a
dose repeated one or several times after a certain time
interval.
[0429] As used herein, a "pharmaceutically appropriate vehicle" is
intended to include any and all carriers, solvents, diluents,
excipients, adjuvants, dispersion media, coatings, antibacterial
and antifungal agents, and absorption delaying agents compatible
with pharmaceutical administration. The composition of the
invention can be in various forms, e.g. solid, liquid or frozen.
Solid (e.g. dry powdered or lyophilized) compositions can be
obtained by a process involving vacuum drying and
freeze-drying.
[0430] Suitably, the pharmaceutical composition of the invention
comprises a diluent appropriate for human or animal use. It is
preferably isotonic, hypotonic or weakly hypertonic and has a
relatively low ionic strength. Representative examples include
sterile water, physiological saline (e.g. sodium chloride),
Ringer's solution, glucose, trehalose or saccharose solutions,
Hank's solution, and other aqueous physiologically balanced salt
solutions (see for example the most current edition of Remington:
The Science and Practice of Pharmacy, A. Gennaro, Lippincott,
Williams & Wilkins). The composition of the invention is
suitably buffered in order to be appropriate for human use at a
physiological or slightly basic pH (e.g. between about pH 7 to
about pH 9). Suitable buffers include without limitation phosphate
buffer (e.g. PBS), bicarbonate buffer and/or Tris buffer.
[0431] The pharmaceutically appropriate vehicles included in the
composition of the invention must also permit to preserve its
stability under the conditions of manufacture and long-term storage
(i.e. at least one month) at freezing (e.g. -70.degree. C.,
-20.degree. C.), refrigerated (e.g. 4.degree. C.) or ambient
temperatures. In this respect, formulations which are particularly
adapted to the composition of the invention include: [0432] 1M
saccharose, 150 mM NaCl, 1 mM MgCl.sub.2, 54 mg/l Tween 80, 10 mM
Tris pH 8.5 (especially when the active ingredient is an adenoviral
vector), [0433] 10 mg/ml mannitol, 1 mg/ml HSA, 20 mM Tris, pH 7.2,
and 150 mM NaCl, and [0434] physiological saline.
[0435] In addition, the composition of the invention may comprise
one or more adjuvant(s) suitable for systemic or mucosal
application in humans. Preferably, the adjuvant is capable of
stimulating immunity to the composition of the invention,
especially a T cell-mediated immunity e.g. through the toll-like
receptors (TLR). Suitable adjuvants are well known in the art.
Representative examples include without limitation alum, mineral
oil emulsion such as Freunds complete and incomplete (IFA),
lipopolysaccharide (Ribi et al., 1986, Immunology and
Immunopharmacology of Bacterial Endotoxins, Plenum Publ. Corp., NY,
p407-419), saponins (Sumino et al., 1998, J. Virol. 72, 4931-4939;
WO 98/56415), imidazo-quinoline compounds (Suader, 2000, J. Am Acad
Dermatol. 43, S6-S11; Smorlesi, 2005, Gene Ther. 12, 1324-1332),
and cationic peptides (Kritsch et al., 2005, J. Chromatogr Anal.
Technol Biomed Life Sci 822, 263-270).
[0436] The composition of the present invention may be administered
by a variety of modes of administration, including systemic,
topical and localized administration. Injection can be performed by
any means, for example by subcutaneous, intradermal, intramuscular,
intravenous, intraperitoneal, intratumoral, intravascular,
intraarterial injection or by direct injection into an artery (e.g.
by hepatic artery infusion) or a vein feeding liver (e.g. injection
into the portal vein). Injections can be made with conventional
syringes and needles, or any other appropriate devices available in
the art. Topical administration can also be performed using
transdermal means (e.g. patch and the like). Administration via
mucosal route can also be envisaged (e.g. intranasal,
intravaginal). In the context of the invention, intramuscular and
subcutaneous administrations constitute the preferred routes.
[0437] The pharmaceutical composition of the invention may be
employed in methods for treating a variety of diseases and
pathologic conditions, especially those associated with an HCV
infection. As used herein, the term "treatment" or "treating"
encompasses prophylaxis and/or therapy. It is especially useful for
treating HCV persistent infection and liver cancer in HCV-infected
patients. The term "cancer" encompasses any cancerous conditions
including diffuse or localized tumors, metastasis, cancerous polyps
as well as preneoplastic lesions (e.g. cirrhosis). Preferably, upon
introduction into a host organism according to the modalities
described herein, the composition of the invention provides a
therapeutic benefit to the treated host. The term "host organism"
is intended to encompass mammals including humans infected with
HCV. The therapeutic benefit can be evidenced by a number of ways,
for instance a decrease of HCV viremia detected in blood, plasma or
sera of an infected mammal as compared to before treatment, and/or
by the detection of an anti-HCV immune response (e.g. production of
anti-HCV antibodies and/or T cell-mediated immunity) and/or by a
modification of the proteomic profile analyzed before and after
therapy (e.g. using the Gene Array Affimetrix technology on blood
cells) and/or by the delay of the symptoms associated with an HCV
infection (e.g. delay in the development of liver cirrhosis or
cancer), and/or by a decrease of liver
inflammation/steatosis/fibrosis conditions typically associated
with HCV infection or by an improved response of the mammal to
conventional therapies.
[0438] The invention also relates to a diagnostic composition for
the detection and/or quantification of the hepatitis C virus
comprising at least two different peptides chosen from the A to D
peptides as defined previously, or at least one B' or C' epitope as
defined previously, or at least one antibody as defined
previously.
[0439] There also, a person skilled in the art will easily
determine the quantity of peptides to be used as a function of the
diagnostic technique used.
[0440] The invention also relates to a process for detection and/or
quantification of the hepatitis C virus in a biological sample
taken from an individual capable of being infected by said virus,
such as plasma, serum or tissue, characterized in that it comprises
stages consisting of: [0441] bringing said biological sample into
contact with at least one of the antibodies of the invention under
conditions allowing the formation of a complex between the virus
and the antibody, [0442] detecting and/or quantifying the formation
of said complex by any appropriate means.
[0443] The processes of detection and/or quantification of the
virus are implemented using standard techniques well known to a
person skilled in the art and there can be mentioned, by way of
illustration, blots, so-called sandwich techniques, competition
techniques and PCR detection techniques, in particular so-called
"real-time" techniques.
[0444] The invention also relates to the use of the compositions of
the invention for the in vitro diagnosis of the hepatitis C virus
in a biological sample.
[0445] Finally, the invention relates to the use of the
compositions of the invention for the preparation of a vaccine
composition.
[0446] In a preferred embodiment, the present invention relates to
the use of the peptide composition, fusion peptide, nucleotide
sequence, vector, host cell or pharmaceutical composition of the
invention for the preparation of a drug intended for inhibiting or
treating or preventing HCV infections, HCV-associated diseases and
pathologic conditions, in an animal, preferably human, according to
the modalities described above.
[0447] The present invention also provides a method for the
treatment or prevention of a human or animal organism, comprising
administering to said organism a therapeutically effective amount
of the peptide composition, fusion peptide, nucleotide sequence,
vector, host cell or pharmaceutical composition of the invention.
If desired, the method of the invention can be carried out in
conjunction with one or more conventional therapeutic modalities
(e.g. radiation, chemotherapy antiviral treatment and/or surgery).
The use of multiple therapeutic approaches provides the patient
with a broader based intervention.
[0448] The method or use of the invention may be carried out
according to a prime boost therapeutic modality which comprises
sequential administration of one or more primer composition(s) and
one or more booster composition(s). Typically, the priming and the
boosting compositions use different vehicles which comprise or
encode at least an antigenic domain in common. In the context of
the invention, a composition of the invention may be used to either
prime or boost or both prime and boost an anti-HCV immune response.
For illustrative purposes, one may prime with a fusion peptide
having the amino acid sequence shown in SEQ ID NO: 234 and boost
with a vector encoding an antigenic domain in common with the
composition of the invention (e.g. an antigenic domain of NS3, NS4
and/or NS5B polypeptide). The source of such material is wide and
includes without limitation peptides, proteins, viral vector from a
variety of viruses, plasmid DNA, proteinaceous particles such as
virus-like particles (Burns et al., 1994, Mol. Biol. Techno. 1:
137-145), cellular materials such as irradiated cells, virus
particles (e.g. as described in WO2004/111082), etc. Alternatively,
one may prime with the adenovirus particles of WO2004/111082 and
subsequently boost with a MVA vector encoding the fusion peptide as
defined in SEQ ID NO: 234. However other prime boost combinations
are also possible in the context of the invention.
[0449] The present invention also provides a method of inducing or
stimulating an immune response against HCV in a host organism
comprising administering to said organism the peptide composition,
fusion peptide, nucleotide sequence, vector, host cell, antibodies
or pharmaceutical composition of the invention so as to induce or
stimulate said immune response. The immune response can be a
specific and/or a nonspecific, a humoral and/or a cell-mediated
response. The immune response is preferably a T cell-mediated
response directed to a T cell-recognized HCV epitope. The ability
of the method of the invention to induce or stimulate an anti-HCV
immune response upon administration in an animal or human organism
can be evaluated either in vitro or in vivo using a variety of
assays which are standard in the art (see for example Coligan et
al., 1992 and 1994, Current Protocols in Immunology; ed J Wiley
& Sons Inc, National Institute of Health). Measurement of
cellular immunity can be performed by measurement of cytokine
profiles secreted by activated effector cells including those
derived from CD4+ and CD8+ T-cells (e.g. quantification of IL-10 or
IFNg-producing cells by ELIspot), by determination of the
activation status of immune effector cells (e.g. T cell
proliferation assays by a classical [.sup.3H] thymidine uptake), by
assaying for antigen-specific T lymphocytes in a sensitized subject
(e.g. peptide-specific lysis in a cytotoxicity assay). The ability
to stimulate a humoral response may be determined by antibody
binding and/or competition in binding (see for example Harlow,
1989, Antibodies, Cold Spring Harbor Press). The method of the
invention can also be further validated in animal models challenged
with an appropriate infectious or tumor-inducing agent (e.g.
surrogate murine challenge assays described in the art using for
example a recombinant Listeria expressing NS3, a NS2-NS5 vaccinia
virus) to determine neutralization of the infectious or
tumor-inducing agent and eventually partial resistance to the
associated symptoms, reflecting an induction or an enhancement of
an anti-HCV immune response. Testing and validation of the
compositions of the invention are also illustrated in the appended
Example section.
[0450] The invention has been described in an illustrative manner,
and it is to be understood that the terminology which has been used
is intended to be in the nature of words of description rather than
of limitation. Obviously, many modifications and variations of the
present invention are possible in light of the above teachings. It
is therefore to be understood that within the scope of the appended
claims, the invention may be practiced in a different way from what
is specifically described herein.
[0451] All of the above cited disclosures of patents, publications
and database entries are specifically incorporated herein by
reference in their entirety to the same extent as if each such
individual patent, publication or entry were specifically and
individually indicated to be incorporated by reference.
[0452] The present invention will be better understood using the
following examples given only by way of illustration and
non-limitatively, as well as using the attached FIGS. 1 to 16, in
which:
[0453] FIG. 1A represents the average production of gamma
interferon for 3 patients (Pt 1 to 3) infected by HCV strains of
genotype 1 and having an HLA. A2+. This production is determined by
the number of spots observed per million of blood cells brought
into contact with the FLAT (FLATCVNGV) and LLG (LLGCIITSL)
peptides,
[0454] FIG. 1B represents the average production of interleukin-10
for 3 patients (Pt 1 to 3) infected by HCV strains of genotype 1
and having an HLA. A2+. This production is determined by the number
of spots observed per million of blood cells brought into contact
with the FLAT and LLG peptides,
[0455] FIG. 2 represents the alignment of the A peptides of the
invention,
[0456] FIG. 3 represents the alignment of the B peptides of the
invention,
[0457] FIG. 4 represents the alignment of the C peptides of the
invention and
[0458] FIG. 5 represents the alignment of the D peptides of the
invention.
[0459] FIG. 6 relates to the A peptide and shows a graph giving the
specific lysis percentage of splenocytes of HLA-A2 transgenic mice
according to the CTL test conditions after immunization of the mice
by the A peptide of the invention,
[0460] FIGS. 7A and 7B relate to the B peptide and show on the one
hand a graph giving the specific lysis percentage of splenocytes of
HLA-A2 transgenic mice according to the conditions of the CTL test
after immunization of the mice by the B peptide of the invention
(FIG. 7A) and on the other hand another graph giving the number of
gamma-interferon-producing cells revealed by the ELISPOT method
originating from the same transgenic mice and brought into contact
with the ATL epitope of the prior art contained in the A peptide
(FIG. 7B),
[0461] FIGS. 8A and 8B relate to the C peptide and show on the one
hand a graph giving the specific lysis percentage of splenocytes of
HLA-A2 transgenic mice according to the CTL test conditions after
immunization of the mice by the C peptide of the invention (FIG.
8A) and on the other hand another graph giving the number of
gamma-interferon-producing cells revealed by the ELISPOT method
originating from the same transgenic mice and brought into contact
with the C peptide (FIG. 8B),
[0462] FIGS. 9A and 9B relate to the D peptide and show on the one
hand a graph giving the specific lysis percentage of splenocytes of
HLA-A2 transgenic mice according to the CTL test conditions after
immunization of the mice by the D peptide of the invention (FIG.
9A) and on the other hand another graph giving the number of
gamma-interferon-producing cells revealed by the ELISPOT method
originating from the same transgenic mice and brought into contact
with the epitope ALY of the prior art contained in the D peptide
(FIG. 9C), and
[0463] FIG. 10 is a graph giving the specific lysis percentage of
splenocytes of HLA-A2 transgenic mice according to the CTL test
conditions after immunization of the mice by the D peptide of the
invention or after immunization with the ALY epitope,
[0464] FIG. 11 is a graph representing the number of
gamma-interferon-producing cells revealed by the ELISPOT method
originating from an HCV-seropositive patient and brought into
contact with the A, B, C and D peptides, as well as with the
compositions A/B, B/C/D and A/B/C/D,
[0465] FIG. 12 is a graph representing the number of
gamma-interferon-producing cells revealed by the ELISPOT method
originating from another HCV-seropositive patient and brought into
contact with the A, B, C and D peptides, as well as with the
compositions A/B, A/C, C/D, A/B/C, A/C/D, B/C/D and A/B/C/D,
[0466] FIG. 13 is a graph representing the number of
gamma-interferon-producing cells revealed by the ELISPOT method
originating from another HCV-seropositive patient and brought into
contact with the A, B, C and D peptides, as well as with the
composition A/B/C/D,
[0467] FIG. 14 is a graph representing the number of
gamma-interferon-producing cells revealed by the ELISPOT method
originating from another HCV-seropositive patient and brought into
contact with the A, B, C and D peptides, as well as with the
compositions A/B, B/D, B/C/D and A/B/C/D.
[0468] FIGS. 15A and 15B illustrate the number of
gamma-interferon-producing cells revealed by ELISPOT of splenocytes
collected from HLA-A2 transgenic mice immunized with DNA plasmids
encoding the B/A/C/D fusion peptide of the invention (FIG. 15A) or
immunized with adenoviruses encoding the B/A/C/D fusion peptide of
the invention (FIG. 15B) using the indicated peptides (GLL, ALY and
CVN). Striped histograms represent polyEp-WT-expressing vectors,
black histograms represent polyEp-C-expressing vectors, stippled
histograms represent polyEp-E3-expressing vectors and white
histograms represent empty vectors. In FIG. 15A, WT, C, E3 and D
represent mice immunized with gwiz plasmids encoding polyEp-WT,
polyEp-C, polyEp-E3 and with an empty gwiz plasmid whereas in FIG.
15B, WT, C, E3 and D represent mice immunized with Ad-polyEp-WT,
Ad-polyEp-C, Ad-polyEp-E3 and with an empty adenovirus vector
respectively
[0469] FIGS. 16A and 16B illustrate the specific lysis percentage
revealed by CTL assay of splenocytes collected from HLA-A2
transgenic mice immunized with gwiz plasmids encoding polyEp-WT,
Ad-polyEp-C, Ad-polyEp-E3 encoding the B/A/C/D fusion peptide of
the invention (FIG. 16A) or immunized with Ad-polyEp-WT,
Ad-polyEp-C and Ad-polyEp-E3 (FIG. 16B). Striped histograms
represent polyEp-WT-expressing vectors, black histograms represent
polyEp-C-expressing vectors, stippled histograms represent
polyEp-E3-expressing vectors and white histograms represent empty
vectors. Tested peptides are indicated under each graph.
Effector/Target (E/T) ratio is 100/1 except for CVN and GLL
peptides where the E/T ratio is 4/1.
EXAMPLE 1
Absence of Correlation Between the Binding Score and the
Immunogenic Power of a Peptide-Epitope
[0470] 1a: Test on Human Cells
[0471] The ability of blood cells from 3 patients infected by the
hepatitis C virus of genotype 1 and having an HLA of type HLA-A2+
to produce gamma-interferon cytokines and interleukin-10, in
response to two peptides predicted by software developed by
bioMerieux (Centre d'Immunologie de Pierre Fabre) to bind to the
HLA-A2 molecule was tested.
[0472] The assay of these cytokines reflects the ability of the
peptides to induce immune responses to cell mediation, either of
type 1 (gamma interferon), or of type 2 (interleukin-10)
[0473] In order to do this, the FLAT (FLATCVNGV) and LLG
(LLGCIITSL) epitopes included in the B peptide of the invention,
which have equivalent binding scores (694 for FLAT and 619 for LLG)
were used.
[0474] Mononucleated blood cells from 3 infected patients (200,000
cells per well) were incubated in an ELISPOT plate (Human IL10
Elispot Set, San Diego, Calif., USA) on which biotinylated
antibodies specific to gamma interferon (BD kit) or interleukin-10
(purified mouse anti-human .gamma.-IFN monoclonal antibody, BD, No.
554548) had been previously fixed, in the presence of the FLAT and
LLG peptides.
[0475] After 48 hours of incubation at 37.degree. C., a period
during which the cells specific to the peptides will locally
produce cytokines which will bind to the specific antibodies,
anti-gamma interferon or anti-interleukin-10 antibodies, alkaline
phosphatase coupled to avidine and the alkaline phosphatase
substrate (NBT/BCIP) were added.
[0476] The violet-blue spots, which represent each gamma-interferon
or interleukin-10-producing cell were counted, using Zeiss's
ELISPOT automated reader (KS Elispot) and only considered positive
when the number of spots per well was greater than 20.
[0477] The results obtained are shown FIG. 1, in which Pt
represents patient. They show that, despite an equivalent binding
score on the HLA-A2 molecule, these two peptides do not induce
equivalent responses.
[0478] In fact, the FLAT peptide induces little if any production
of the two cytokines in the three patients, whereas the LLG peptide
induces a significant production of gamma interferon in 2 of the 3
patients and of interleukin-10 in the three patients.
[0479] The results obtained also show that the peptides of the
invention, as well as their combination was not evident with regard
to the teachings of the prior art.
[0480] 1b: Test on Murine Cells
[0481] Mice transgenic for the molecule HLA.A2.1 and devoid of
murine class I molecules (Pascolo S., et al., 1997, J. Exp Med.,
185: 2043-2051) were immunized with the FLL and ILA epitopes
predicted as being class I epitopes restricted by the molecule
HLA.A2.1. The FLL peptide has an average score (515) whereas the
ILA peptide has a very high score (893).
[0482] In order to do this, the mice were immunized in the base of
their tails with a peptide mixture containing one of the above
epitopes (60 .mu.M) and the T-helper peptide (Lone, Y. C. et al.,
(1998), J. Immunother. 21:283-294) (60 .mu.M) emulsified in
incomplete Freund's adjuvant (Sigma St Louis, Mo.). The mice
received two injections with an interval of two weeks and the
immune responses were analyzed two weeks after the last
immunization as indicated in Example 1 (Elispot).
[0483] The results are shown in Table 1 below. TABLE-US-00001 TABLE
1 Viral Number of spots/10.sup.6 Epitope antigen Score cells.sup.1
* FLLLADARV E2 515 333; 215; 622 ILAGYAGAGV NS4 893 5; 2; 0
.sup.1Gamma-interferon-producing cells * Number of spots for three
individual mice tested.
[0484] These results show that the score ascribed to a peptide does
not reflect its immunogenicity.
EXAMPLE 2
Demonstration of the Immunogenicity of the Peptides/Peptide
Compositions/Epitopes of the Invention in Mice
[0485] HLA-A2.1-transgenic mice were immunized either with the A
peptide of sequence SEQ ID NO: 3, or with the B peptide of sequence
SEQ ID NO: 47, or with the C peptide of sequence SEQ ID NO: 129, or
with the D peptide of sequence SEQ ID NO: 176, or with the B'
epitope of sequence SEQ ID NO: 214 or with the ATL and ALY epitopes
of the prior art contained respectively in the A and D peptides
(ATLGFGAYM, amino acids 1260-1268 of the viral polyprotein and
ALYDVVSTL, amino acids 2594-2602 of the viral polyprotein).
[0486] In order to do this, 2 sub-cutaneous injections were
administered into the base of the tail of each mouse, with an
interval of 15 days, of a mixture containing 18 nmoles of peptide
of the invention and 18 nmoles of the HBV core helper peptide in
incomplete Freund's adjuvant (IFA, Brinster et al., Hepatology
2001) or containing 60 nmoles of epitope of the invention and 60
nmoles of the helper peptide in IFA.
[0487] The control mice received only the helper peptide in IFA.
Fifteen days after the 2nd injection, the cell response was
analyzed by isolating the spleen cells (splenocytes) of mice and a
CTL test and an ELISPOT test were carried out as follows:
[0488] For the CTL test, these splenocytes were cultured in 24-well
plates in the presence of 5 .mu.M of the peptide or epitope of
interest and 10 U of recombinant murine interleukin-2 (Brinster et
al., Hepatology 2001) per ml in alpha minimum essential medium
(.alpha.AMEM) for 5 days. On the 5th day, the restimulation stage
was carried out, consisting of adding to the splenocytes in culture
splenocytes of naive mice in the presence of the peptide or epitope
of interest over 2 days. On the 7th day, the CTL test itself was
carried out, consisting of bringing together the splenocytes of the
immunized mice after the 7 days of culture (effective cells) and
EL4 S3-Rob HDD cells charged with 10 .mu.M of the peptide or
epitope of interest marked with Cr.sup.51 (target cells). The
specific cytotoxic activity of the effective cells was determined
by measuring, after 4 hours of incubation with the target cells,
the Cr.sup.5' released following the lysis of the target cells
using a .gamma.-Cobra II (Packard, Rungis, France) counting device.
The spontaneous and maximum release from wells containing either
the medium alone, or the lysis buffer (HCl 1N) were determined. The
specific percentage of cytotoxicity was calculated by the formula:
(release in the test-spontaneous release)/(maximum
release-spontaneous release).times.100. The peptide or
epitope-specific lysis was determined by the difference between the
percentage of specific lysis obtained in the presence or absence of
the peptide or epitope.
[0489] The ELISPOT test was carried out by culturing the
splenocytes for 48 hours in Multiscreen (Millipore) 96-well plates
previously coated with anti-.quadrature.-interferon (.gamma.IFN)
antibodies (10 .mu.g/ml final). The splenocytes were cultured in
the presence of 10 .mu.M of peptide or epitope of interest and 10 U
of recombinant murine interleukin-2 per ml in .alpha.MEM. For the
positive control, the splenocytes were cultured in the presence of
concanavalin A (5 .mu.g/ml). For the negative control, the
splenocytes were cultured either in the presence of a non-specific
peptide belonging to the HCV capsid protein, of sequence DLMGYIPLV
(also called irrelevant peptide), or in medium alone without
peptide or epitope of interest. The wells were washed three times,
respectively with PBS-Tween 0.05% then PBS, an operation followed
by incubation for 2 hours with biotinylated anti-.gamma.-IFN
antibodies of mice. After washing, the wells were incubated for 1
hour with a streptavidin-horseradish peroxidase conjugate and the
enzymatic activity was revealed by degradation of the AEC
(aminoethylcarbazole) substrate. The spots obtained were counted
using a Zeiss ELISpot reader (Zeiss microscope coupled with the
KS-ELISpot software).
[0490] The results of the CTL tests are shown in FIGS. 6, 7A to 9A
and 10 where S1 is mouse 1, S2 is mouse 2, S3 is mouse 3, S4 is
mouse 4 and S neg is the control mouse and where: [0491] FIG. 6
shows the specific lysis percentage, as a function of the
effectors/target ratio, after injection of the A peptide taking the
ATL epitope as target, [0492] FIG. 7A shows the specific lysis
percentage, as a function of the effectors/target ratio, after
injection of the B peptide taking the B' epitope as target, [0493]
FIG. 8A shows the specific lysis percentage, as a function of the
effectors/target ratio, after injection of the C peptide taking the
C peptide as target, [0494] FIG. 9A shows the specific lysis
percentage, as a function of the effectors/target ratio, after
injection of the D peptide taking the ALY epitope as target, and
[0495] FIG. 10 shows the specific lysis percentage, as a function
of the effectors/target ratio, after injection of the D peptide
taking the ALY epitope (mice S1 and S3) as target or the ALY
epitope taking the ALY epitope (mice S3 and S4) as target.
[0496] The results as shown in FIGS. 6 to 9 show that the injection
of each peptide induces a cytotoxic response against the
corresponding epitopes such that:
[0497] (i) both said peptides and said epitopes have an immunogenic
power, and
[0498] (ii) the peptides of the invention are capable of inducing
immune responses specific to epitopes present in the natural
infection.
[0499] It is to be noted that FIG. 10 shows that the D peptide and
the epitope have identical lysis effectiveness. It is therefore
clear from this experience, with regard to the quantities of
peptide and epitopes used (injection of 18 nmoles of the D peptide
(mice S1 and S2) and injection of 60 nmoles of the ALY epitope
(mice S3 and S4), taking the ALY epitope as target), that the
peptides of the invention have the advantage that they are
immunogenic at doses lower than the epitopes that they contain.
[0500] The CTL test was also repeated injecting the composition
ABCD or the B peptide taking the B' epitope as target and injecting
the composition ABCD or the C peptide taking the C peptide as
target. The results are indicated in Table 3 below. TABLE-US-00002
TABLE 3 Effectors/targets ratio 100:1 33:1 Injection ABCD Target B'
S1 55 52 S2 32 55 S3 53 65 Injection B S1 46 63 S2 22 27 Injection
ABCD Target C S1 52 69 S2 1 67 Injection C S1 2 9 S2 0 0
[0501] The results in the above table demonstrate that injection of
the combination of the 4 A, B, C and D peptides induces a cytotoxic
response more effective than injection of the peptides alone.
[0502] The results of the ELISPOT tests are shown in FIGS. 7B to 9B
where S1, S2 S3 and S neg have the same definitions as previously
and where: [0503] FIG. 7B shows the number of spots relative to
10.sup.6 cells for mice having received the B peptide relative to
the B' epitope target, as well as the number of spots for the
irrelevant peptide. [0504] FIG. 8B shows the number of spots
relative to 10.sup.6 cells for mice having received the C peptide
relative to the C peptide target, as well as the number of spots
for the irrelevant peptide. [0505] FIG. 9B shows the number of
spots relative to 10.sup.6 cells for mice having received the D
peptide relative to the ALY epitope target, as well as the number
of spots for the irrelevant peptide.
[0506] The results shown in FIGS. 7B to 9B confirm the good
immunogenic power of the peptides and epitopes of the
invention.
EXAMPLE 3
Demonstration of the Immunogenicity of the Peptides/Peptide
Compositions of the Invention in Humans
[0507] Peripheral blood mononucleated cells from four patients
chronically infected by the hepatitis C virus were purified by
Ficoll gradient centrifugation. Two hundred thousand cells were
pre-incubated at 37.degree. C., 5% CO.sub.2, overnight in
polypropylene tubes in the presence of the A to D peptides, having
the amino acid sequences as defined in Example 2 above, and their
compositions, in culture medium comprising RPMI1640 (Invitrogen
Life technology, Cergy Pontoise, France) supplemented with 2mM of
L-Glutamine (Invitrogen Life Technology), 50 Ul/ml of penicillin
(Invitrogen Life Technology), 50 .mu.g/ml of streptomycin
(Invitrogen Life Technology) and 10% of f tal calf serum (Hyclone,
Logan, Utah, USA) according to the following Table 4:
TABLE-US-00003 TABLE 4 Clinical Peptides/peptide Patient Genotype
status compositions 1 1b Not treated A, B, C, D, AB, BCD, ABCD 2 1
Not responsive to A, B, C, D, AB, AC, CD, ribavirin M12 ABC, ACD,
BCD, ABCD 3 4f Not treated A, B, C, D, ABCD 4 1 Not responsive to
A, B, C, D, AB, BD, BCD, .gamma.IFN + ABCD ribavirin M6
[0508] The peptides, alone or in combination, were present in the
following concentrations: A at 10 .mu.M, B at 5 .mu.M, C at 1 .mu.M
and D at .mu.M.
[0509] The cells were then transferred to an ELISPOT plate made of
PVDF (polyvinylidene fluoride) previously coated with
anti-.gamma.IFN antibodies according to the manufacturer's
(Diaclone, Besangon, France) recommendations and incubated for
another 24 hours at 37.degree. C., 5% CO.sub.2. As previously, here
cytokine-producing cells are sought, which will form blue spots
which are revealed, after sequential incubation with a biotinylated
anti-.gamma.IFN antibody and PAL coupled to streptavidin, by
degradation of the BCIP/NBT (salt of
5-bromo-4-chloro-3-indolylphosphate p-toluidine/nitroblue
tetrazolium chloride) substrate and which are counted using the
Zeiss ELISPOT reader.
[0510] As positive control, the peptides/peptide compositions were
replaced by tetanic toxin (TT) at 1 .mu.g/ml.
[0511] This procedure was repeated starting with cells from
HCV-seronegative patients.
[0512] The results are shown in FIGS. 11 to 14 giving the number of
spots per million cells as a function of the peptides and their
combinations (average of the triplicates) obtained with patients 1
to 4 respectively, and in which the dotted line represents the
significance threshold of the test (50 spots), Pt is patient and
HCVneg is an HCV-seronegative patient.
[0513] The results demonstrate that the peptide compositions of the
invention have a strong immunogenic power.
EXAMPLE 4
Minigenes Encoding a Chimeric Polypeptide Composed of the Peptides
of the Invention in Fusion Induced T Cell Response in
HLA-A2.1-Transgenic Mice
[0514] Material and Methods
[0515] Peptides
[0516] The synthetic peptides used for Elispot and CTL assays were
purchased from Eurogentec (Liege, Belgium). Seven peptides were
generated with a purity of more than 85%, which sequences derived
from the HCV-JA strain (Kato et al., 1990, Proc. Natl. Acad. Sci.
USA 87: 9524-9528): ATLGFGAYM, GLLGCIITSL, CVNGVCWTV, LLFNILGGWV,
SLMAFTASI, ALYDVVSTL, and RLIVFPDLGV, respectively referred as to
ATL, GLL, CVN, LLF, SLM, ALY and RLI. They were dissolved in 100%
DMSO at 10.mu.M and stored at -20.degree. C. until use.
[0517] Cell Cultures
[0518] Human Huh 7 hepatoma cells were maintained at 37.degree. C.
in 5% CO.sub.2 atmosphere in complete DMEM medium containing
Dulbecco's modified Eagles medium supplemented with 10% fetal calf
serum, 2 mM L-Glutamine and 100UI/ml penicillin/streptomycin
(Sigma). HeLa cells (available at ATCC), a human cervix carcinoma
cell line, were cultured according to supplier's instructions.
[0519] Mice
[0520] The HLA-A2.1 transgenic mice used for vaccination
experiments are described in Pascolo et al. (1997, J. Exp. Med.
185: 2043-2051). These mice express a transgenic monochain
histocompatibility class I molecule in which the C-terminus of the
human .beta.2-m is covalently linked to the N-terminus of a
chimeric heavy chain composed of the HLA-A0201 .alpha.1 and
.alpha.2 and H2-D.sup.b .alpha.3-transmembrane and intracytoplasmic
domains. In addition, they are knocked out for the H2-D.sup.b and
murine .beta.2-m genes which were disrupted by homologous
recombination. Mice were hosted in appropriate animal care
facilities and handled following international guidelines required
for experiments with animals.
[0521] Plasmids and Adenovirus Constructions
[0522] Three synthetic oligonucleotidic sequences were synthesised
by Geneart (Regensburg, Germany) and cloned in pCR-Script Amp
plasmids-(Stratagene, La Jolla, US). The polyEp-WT minigene
comprised a fusion of the wild-type nucleotide sequences (as found
in the HCV JA genome) encoding B/A/C/D peptides. The polyEp-C
minigene comprised a B/A/C/D encoding nucleotide sequence which
codons had been optimized for translation in mammalian cells by
computer software. The polyEp-E3 minigene encodes the B/A/C/D
fusion with an N-terminal adenovirus E3 leader peptide
(RYMILGLLALAAVCSA; SEQ ID NO: 233) for endoplasmic reticulum
targeting.
[0523] The synthetic minigenes were amplified by PCR using
appropriate primers incorporating a Kozack sequence and an AUG
codon. The amplified sequences were inserted between the SalI and
NotI sites of the gwiz plasmid (Gene Therapy System, San Diego,
US), and placed under transcriptional control of the CMV promoter.
The resulting gwiz constructions are named polyEp-WT, polyEp-C and
polyEp-E3 respectively. A myc-tagged version was also generated.
For this purpose, the above-described minigenes were inserted in
the pcDNA3.1/myc-His/lacZ (Invitrogen, Carlsbad, US) in order to
add a myc tag at the carboxy-terminus. The tagged minigenes were
amplified by PCR as described above and inserted between SalI and
NotI sites of the gwiz plasmid.
[0524] The myc-tagged minigenes were also cloned in E1-defective
Ad5 vectors (kindly provided by the Gene Vector Production Network
of Genethon, Paris, France), giving Ad-polyEp-WT, Ad-polyEp-C and
Ad-polyEp-E3. As in the gwiz plasmids, the minigenes were placed
under transcriptional control of the CMV promoter. Adenovirus
stocks were produced in an appropriate packaging cell line such as
293 (Graham et al., 1977, J. Gen. Virol. 36: 59-72) or PERC.6
(Fallaux et al., 1998, Human Gene Ther. 9: 1909-1917) cells.
[0525] DNA Transfection and Adenovirus Infection
[0526] Human Huh7 cells were transfected with the different plasmid
constructions described above in the presence of Lipofectamin/Plus
Reagent (Invitrogen, Carlsbad, US). Briefly, 5.times.15 cells were
plated in 6-well plates and 24 h later 1 .mu.g of DNA was added to
each well with Lipofectamin and Plus Reagent according to the
manufacturer's instructions. HeLa or Huh7 cells were infected with
the adenoviral vectors encoding the different myc-tagged minigenes
at a MOI of 100. Briefly, 5.times.10.sup.5 cells were plated in
6-well plates and 24 h later 5.times.10.sup.7 iu of each adenovirus
construction were added to each well.
[0527] Western Blot
[0528] Huh7 cells transfected by gwiz plasmids or infected with the
different adenoviruses were incubated in ice-cold lysis buffer
containing 1% NP40, 8M of urea and complete protease inhibitor
(Roche Meyland). Lysed cells were centrifuged at 12,000 g for 10
min, and supernatants containing cytosolic fractions were assayed
for protein concentrations (Comassie Blue, Pierce, Rockford, US) as
described by the manufacturer. Equivalent protein amounts were
loaded on a 12% acrylamide gel and run for 2 h. After
electrophoretic transfer, polyvinylidene fluoride membranes (Hybond
PVDF transfer membrane, Amersham, Uppsala, Sweden) were saturated
and incubated with an anti-myc monoclonal antibody (dilution 1/5
000, Invitrogen, Carlsbad, US) and an anti-actin moclonal antibody
(dilution 1/10 000, Sigma, Saint-Louis, US). Membranes were then
incubated with an HRP-conjugated goat anti-mouse IgG antibody (1/10
000, Dako Cytomation, Glostrup, Denmark) and revealed by
chemiluminescence (ECL Western-blotting Detection System,
Amersham).
[0529] Immunofluorescence
[0530] Huh7 or HeLa cells were cultured and transfected or infected
on coverslips in 6-well plates as previously described. 48 h post
transfection or infection, cells were washed in 1.times.PBS, fixed
with 1.times.PBS plus 4% paraformaldehyde followed by 1.times.PBS
plus Glycine 0,1M and permeabilized with 1.times.PBS plus Triton
0,1%. Staining was carried out by sequential incubation with first
anti-myc monoclonal antibody (mouse IgG, dilution 1/1000,
Invitrogen, Carlsbad, US) or anti-calnexin monoclonal antibody
(rabbit IgG, dilution 1/500, Stressgen, San Diego, US) for cellular
localization experiments and then TRITC-conjugated goat anti-mouse
IgG antibody (1/150, Sigma Aldrich) or FITC-conjugated swine
anti-rabbit IgG antibody (dilution 1/40, Dako Cytomation). Antibody
incubations and washes were done in PBS 1.times. plus BSA 0.1%.
Cell nucleus were counter stained with Hoechst reagent (Sigma) at
1/200 diluted in PBS 80%-Glycerol 20%, observed using a
fluorescence microscope (Zeiss, Le Pecq, France) and analysed with
Adobe Photoshop 5.0.
[0531] Immunization Protocols
[0532] All plasmid DNA preparations were generated using
endotoxin-free Macherey-Nagel purification columns (Macherey-Nagel,
Duren, Germany) and adenovirus stocks were tested for the absence
of endotoxins (Genethon, Paris, France). Six to eight week-old
HLA-A2.1 transgenic mice were immunized with two intramuscular
injections at 2-weeks intervals of 100 .mu.g of gwiz plasmid,
polyEp-WT, polyEp-C or polyEp-E3. Adenovirus immunization was
carried out with one intramuscular injection of 10.sup.9 IU of
Ad-polyEp-WTmyc, Ad-polyEp-Cmyc or Ad-polyEp-E3myc. Control mice
were injected with an empty gwiz plasmid or a non-recombinant
adenoviral vector.
[0533] Elispot Assays
[0534] Splenocytes (2.times.10.sup.5 cells per well) collected from
immunized mice were cultured in triplicate for 40 h in Multiscreen
plates (Millipore, France) coated with anti-mouse IFN.gamma.
monoclonal antibody (Pharmingen, France; 10 .mu.g/ml) in complete
.alpha.MEM culture medium (GIBCO BRL, France) in the presence of
recombinant IL-2 (PeproTech Inc, England) at 10 U/mL alone as
negative control, or with 10 .mu.M of peptide (specific or
irrelevant), or 5 .mu.g/mL of Concavalin A as positive control.
IFN.gamma.-producing cells were quantified by cytokine-specific
enzyme linked immunospot assay (ELISPOT) as previously described
(Himoudi et al., 2002, J. Virol 76: 12735-12746). Each spot
represents an individual IFN.gamma.-producing cell. The number of
spots in negative control was subtracted to the number of spots
obtained in the experimental wells containing specific or
irrelevant peptide. Results are shown as the mean value obtained
for triplicate wells.
[0535] CTL Assays
[0536] CTL assays were performed as previously described (Brinster
et al., 2001, Hepatology 34: 1206-1217). Briefly, spleen cells were
stimulated for 7 days in the presence of 5 .mu.M peptide and then
re-stimulated on day 5 using irradiated cells loaded with 10 .mu.M
of the selected peptide. On day 7, the re-stimulated spleen cells
were used as effector cells in a standard .sup.51Cr-release assay
against EL4S3-Rob HHD target cells, either loaded or not with 10
.mu.M of the selected peptide. Effector and target cells were also
used at different ratio. After 4 h incubation, .sup.51Cr release
was measured using a TopCount NXT counter (Packard). Spontaneous
and total lysis were determined from wells containing target cells,
loaded or not, either in medium alone or in lysis buffer (HCl 1N)
respectively. Specific cytotoxicity was calculated using the
formula: (release in assay-spontaneous release)/(total
lysis-spontaneous release).times.100. For each effector/target cell
ratio, results are shown as the mean percentage of specific lysis
obtained from duplicate points.
[0537] Listeria Monocytogenes Infectious Challenge A recombinant
and kanamycin-resistant Listeria Monocytogenes expressing the HCV
NS3 protein (named TC-LNS3; Simon et al., 2003, Infect. Immunol.
71, 6372-6380) was inoculated intravenously to immunized mice 15
days after adenovirus immunization (an intramuscular administration
of 10.sup.9 IU/mice as described above). Mice were anaesthetized
and received from 1 to 2 LD50 of TC-LNS3 (1LD50=9.10.sup.7 pfu).
Two days later, mice were sacrified and pieces of liver and spleen
were removed and weighted. The collected organ samples were
homogenized in PBS/Tween 0.05% and residual bacterial titers were
determined by serial dilution on BHI-agar plates (containing 30
.mu.g/mL kanamycin). After 48 to 60 h of culture at room
temperature, colonies obtained on plates were numerated and titers
corresponding to numbers of bacteria per mg of organs were
calculated.
[0538] Results
[0539] Both CD8+ and CD4+ lymphocytes mediated immune responses
play a critical role in the outcome of HCV infection. Examples 2
and 3 demonstrate the capacity of four HCV peptides from
non-structural protein NS3 (A and B peptides), NS4 (C peptide) and
NS5B (D peptide) of inducing T cell responses alone or in various
combinations in natural infection (Example 3) or in a transgenic
mouse model (Example 2).
[0540] The four peptides were combined in a unique chimeric fusion
peptide of 137 amino acids having the amino acid sequence disclosed
in SEQ ID NO: 234 and comprising from N to C terminus B/A/C/ID
peptides (plus an initiator Met). Three minigene constructs were
generated, respectively polyEp-WT, polyEp-C and polyEp-E3 which
encode the same chimeric B/A/C/D fusion peptide but differ at the
nucleotide level. The polyEp-WT construct comprises the "natural"
nucleotide sequence as found in the HCV-JA genome encoding the 4
peptides fused in the B/A/C/ID combination (SEQ ID NO: 235). In
polyEp-C, the nucleotide sequence encoding each of the peptide has
been optimized for translation in mammalian cells (SEQ ID NO: 236).
The polyEp-E3 construct (SEQ ID NO: 237) encodes the B/A/C/D fusion
preceded at its N-terminus by the leader sequence of the adenovirus
E3 protein for endoplasmic reticulum (ER) targeting of the
resulting 154 amino acid long polypeptide (Met +16 amino acid E3
leader sequence +137 amino acid B/A/C/ID fusion). The minigene
constructs were cloned in gwiz DNA plasmids and E1-defective
adenovirus vectors under control of the CMV promoter in both
cases.
[0541] Expression of the minigene products was evaluated by Western
blot and immunofluorescence in Huh 7 cells transfected or infected
with myc-tagged minigene vectors. Huh 7 cells transfected with an
empty gwiz plasmid or infected with a non-recombinant adenoviral
vector were used as negative controls. Myc-tagged fusion peptides
were revealed using an anti-myc monoclonal antibody with or without
MG132 proteasome inhibitor. Western blot analysis revealed a unique
band having the expected molecular weight of approximately 16 KD
for the fusion peptide encoded by polyEp-WT and polyEp-C and
approximately 17 KD for the fusion peptide encoded by polyEp-E3. As
expected, no proteins were highlighted in Huh 7 negative controls.
Optimization of the minigene codons for mammalian translation does
not result in a significant improvement of expression
(approximately same level of expression in Huh 7 cells transfected
with polyEp-WT plasmid as in Huh 7 cells transfected with
polyEp-C). Interestingly, addition of the ER targeting E3 leader
signal provides a partial protection from proteasome degradation as
illustrated by the fusion peptide detected in the absence of MG132
in samples collected from Huh7 transfected polyEp-E3-encoding
plasmid.
[0542] Immunofluorescence analysis confirmed expression of the
minigene products in Huh 7 and HeLa transfected cells.
Interestingly, fusion peptides encoded by polyEp-WT and polyEp-C
plasmids showed a cytoplasmic localization whereas ER localisation
is suggested for the fusion peptide encoded by polyEp-E3.
[0543] The immunogenic activity of the fusion peptides was
evaluated in HLA-A2.1 transgenic mice immunized by two
intramuscular injections at two weeks interval of 15 plasmids
encoding polyEp-WT, polyEp-C or polyEp-E3 (100
.mu.g/injection/mouse). Specific anti-HCV T cell responses were
analysed 15 days after the last immunization by Elispot and CTL
assays against 7 HLA-A2 restricted epitopes contained in the fusion
peptides, respectively ATL, GLL, CVN, LLF, SLM, ALY and RLI. As
illustrated in FIG. 15A, the minigene-encoding gwiz plasmids are
able to induce IFN.gamma. producing cells specific of 2 out of the
7 tested epitopes, GLL and ALY epitopes. More specifically,
GLL-specific responses ranged from 75 to 212 spots per 10.sup.6
cells for 4 out of 6 mice immunized with polyEp-WT plasmid, from
142 to 153 spots per 10.sup.6 cells for 2 out of 6 mice immunized
with polyEp-C plasmid and from 64 to 118 spots per 10.sup.6 cells
for 3 out of 6 mice immunized with polyEp-E3 plasmid. Specific
responses against ALY epitope are also detected with number of
spots per 10.sup.6 cells ranging from 75 to 215 spots for 2 out of
6 mice for polyEp-WT and from 117 to 130 spots for 2 out of 6 mice
for polyEp-E3. For polyEp-C, only 1 out of 6 mice displayed
ALY-specific IFN.gamma. producing cells (187 spots). As expected no
specific IFN.gamma. producing cells are revealed against the
irrelevant epitope (data not shown). The absence of specific
responses against the 5 other HCV-specific epitopes can be
explained by the fact that these epitopes are usually poor inducers
of IFN.gamma. producing cells in the HLA-A2.1 transgenic mouse
model (Himoudi et al, 2002, J. Virol. 76, 12735-12746; Martin et
al., 2004, J. Med. Virol. 74, 397-405).
[0544] Specific cytolytic responses were also characterized by CTL
assays in HLA-A2.1 transgenic mice twice immunized with
minigene-encoding plasmids. Interestingly, as illustrated in FIG.
16A, cytolytic responses targeted a larger number of epitopes than
IFNg-producing T cells. The polyEp-WT plasmid induced cytolytic
responses specific of 4 epitopes, ATL, GLL, CVN and ALY (with GLL
and ALY-specific lysis higher than 45% in all immunized mice). The
cytolytic responses obtained after immunization with PolyEp-C
plasmid targeted three HCV epitopes, ATL, CVN and ALY with the
stronger responses being generated with CVN (45% to 62% of specific
lysis for 2 out of 3 mice). Immunization with PolyEp-E3 plasmid
induced cytolytic responses specific of ATL, GLL and ALY which are
similar as those obtained after injections of PolyEp-WT plasmid.
Specific lysis higher than 50% was obtained against GLL and ALY in
all immunized animals (from 51% to 77% of specific lysis against
GLL for 3 out of 3 mice and from 50% to 68% of specific lysis
against ALY for 3 out of 3 mice).
[0545] Immunogenicity of minigene-encoded fusion peptide was also
evaluated in transgenic HLA-A2.1 immunized with an intramuscular
injection of Ad-polyEp-WT, Ad-polyEp-C or Ad-polyEp-E3. As
illustrated in FIG. 15B, Elispot assays revealed a strong specific
immune response against GLL, CVN and ALY epitopes. GLL-specific
responses were detected in all vaccinated mice and ranged from 395
to 1988 spots, from 660 to 1978 spots and from 237 to 1504 spots
per 10.sup.6 cells, respectively in Ad-polyEp-WT, Ad-polyEp-C and
Ad-polyEp-E3 immunized mice. CVN-specific IFN.gamma. producing
cells were detected in 2 out of 6 Ad-polyEp-WT injected mice (from
184 to 430 spots per 10.sup.6 cells), and 4 out of 6 Ad-polyEp-E3
injected mice (from 53 to 146 spots per 10.sup.6 cells).
ALY-specific IFN.gamma. producing cells were detected in 3 out of 6
Ad-polyEp-WT injected mice (from 74 to 531 spots per 10.sup.6
cells), and 2 out of 6 Ad-polyEp-E3 injected mice (from 103 to 531
spots per 10.sup.6 cells). Only 1 out of 6 mice immunized with
Ad-polyEp-C displayed IFN.gamma. producing cells specific of CVN
(85 spots/10.sup.6 cells) and of ALY (252 spots/10.sup.6
cells).
[0546] Cytolytic immune responses were measured by CTL assays
against five HLA-A2 epitopes, four low inducer epitopes in DNA
immunization assays (ATL, SLM, LLF and RLI) and a potent inducer
epitope (CVN). After injection of the different minigene-encoding
adenoviruses, no specific cytolytic response could be observed
against SLM and RLI peptides. As illustrated in FIG. 16B, moderate
cytolytic responses were detected against ATL in animals immunized
with Ad-polyEp-WT and Ad-polyEp-E3 (specific lysis ranging from 27%
to 38% for 1 to 2 mice out of 3) whereas no anti-ATL response was
obtained following immunization with Ad-polyEp-C. In contrast, high
CVN-specific cytolytic responses were detected following injection
of minigene-encoding adenoviruses (39% to 46% of specific lysis in
2 out of 3 Ad-polyEp-WT-injected mice; 54% to 63% of specific lysis
in the 3 mice injected with Ad-polyEp-C and 45 to 59% of specific
lysis in the Ad-polyEp-E3-injected mice).
[0547] The protective immunity conferred by the fusion peptides was
evaluated in HLA-A2.1 transgenic mice immunized with the different
minigene-encoding adenoviruses during a recombinant Listeria
surrogate challenge assay. Mice received an intramuscular injection
of Ad-polyEp-WT, Ad-polyEp-C or Ad-polyEp-E3 at Day 0 and were
challenged by intravenous injection of TC-LNS3 15 days later.
Control mice are injected under the same condition with a
non-recombinant (i.e. empty) adenovirus. Mice were sacrified two
days after the challenge and residual bacterial titers were
determined in spleens and livers. The results show that
Ad-polyEp-WT and Ad-polyEp-E3 immunisations allowed a significant
decrease of TC-LNS3 residual titers in spleen as compared to
control mice. Based on the average titers observed for each group,
this decrease is about 1 log for Ad-polyEp-WT and about 2 logs for
Ad-polyEp-E3 as compared to empty adenovirus (p=0.0253 and p=0.0143
respectively, according to Mann Whitney statistical analysis). In
livers, a significant decrease of TC-LNS3 residual titers was
observed only for Ad-polyEp-E3 immunised mice (about 1.5 log of
decrease considering the mean of each group) compared to control
mice immunised with an empty adenoviral vector (p=0.0143 according
to Mann Whitney statistical analysis).
[0548] In conclusion, immunization with DNA plasmids encoding the
B/A/C/D fusion peptide was able to induce both IFN.gamma. producing
cells and cytolytic responses respectively directed against 2 to 4
HLA-A2 restricted epitopes, depending on the constructions and the
assays employed. Equivalent induced T cell responses are observed
for the three minigenes under the experimental conditions tested
and codon optimization did not lead to any improvement.
Interestingly, adenovirus vectorization induced the strongest T
cell reponses. More specifically, immunization with adenovirus
vectors permits to broader spectrum of HLA-A2 restricted epitopes
recognized by IFN.gamma.-producing cells (2 epitopes out of 7
recognized after DNA immunization against 3 after Adenovirus
immunization) and to maximize both the cytolytic and the IFN.gamma.
producing cell-mediated immune responses (higher level of specific
lysis and higher number of spots when the mice are injected with
the minigene-encoding adenoviruses than with DNA plasmids). A
protective immunity was conferred by Ad-polyEp-WT and Ad-polyEp-E3
against a recombinant Listeria expressing the NS3 protein. Based on
these 5 results, vectorized minigenes encoding the B/A/C/D fusion
peptide are potential HCV vaccine candidates.
Sequence CWU 1
1
237 1 31 PRT artificial sequence anti-HCV immunogenic peptide
MISC_FEATURE (1)..(1) Xaa = Y or H MISC_FEATURE (3)..(3) Xaa = A,
G, or T MISC_FEATURE (9)..(9) Xaa = R or L MISC_FEATURE (16)..(16)
Xaa = A or T MISC_FEATURE (20)..(20) Xaa = G or S MISC_FEATURE
(23)..(23) Xaa = A, T, or V MISC_FEATURE (29)..(29) Xaa = H or Y
MISC_FEATURE (31)..(31) Xaa = I, T, M or V 1 Xaa Ala Xaa Gln Gly
Tyr Lys Val Xaa Val Leu Asn Pro Ser Val Xaa 1 5 10 15 Ala Thr Leu
Xaa Phe Gly Xaa Tyr Met Ser Lys Ala Xaa Gly Xaa 20 25 30 2 46 PRT
artificial sequence anti-HCV immunogenic peptide MISC_FEATURE
(2)..(2) Xaa = T or N MISC_FEATURE (3)..(3) Xaa = K or R
MISC_FEATURE (6)..(6) Xaa = A or V MISC_FEATURE (7)..(7) Xaa = A or
E MISC_FEATURE (8)..(8) Xaa = Y or H MISC_FEATURE (10)..(10) Xaa =
A, G, or T MISC_FEATURE (16)..(16) Xaa = R or L MISC_FEATURE
(23)..(23) Xaa = A or T MISC_FEATURE (27)..(27) Xaa = G or S
MISC_FEATURE (30)..(30) Xaa = A, T, or V MISC_FEATURE (36)..(36)
Xaa = H or Y MISC_FEATURE (38)..(38) Xaa = I, T, M or V
MISC_FEATURE (39)..(39) Xaa = E or D MISC_FEATURE (41)..(41) Xaa =
N or S MISC_FEATURE (42)..(42) Xaa = I, L or V MISC_FEATURE
(43)..(43) Xaa = R or S MISC_FEATURE (44)..(44) Xaa = T or S 2 Ser
Xaa Xaa Val Pro Xaa Xaa Xaa Ala Xaa Gln Gly Tyr Lys Val Xaa 1 5 10
15 Val Leu Asn Pro Ser Val Xaa Ala Thr Leu Xaa Phe Gly Xaa Tyr Met
20 25 30 Ser Lys Ala Xaa Gly Xaa Xaa Pro Xaa Xaa Xaa Xaa Gly Val 35
40 45 3 31 PRT artificial sequence anti-HCV immunogenic peptide 3
Tyr Ala Ala Gln Gly Tyr Lys Val Arg Val Leu Asn Pro Ser Val Ala 1 5
10 15 Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly Ile
20 25 30 4 31 PRT artificial sequence anti-HCV immunogenic peptide
4 Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1
5 10 15 Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly Ile
20 25 30 5 31 PRT artificial sequence anti-HCV immunogenic peptide
5 Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1
5 10 15 Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly Val
20 25 30 6 31 PRT artificial sequence anti-HCV immunogenic peptide
6 Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1
5 10 15 Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly Thr
20 25 30 7 31 PRT artificial sequence anti-HCV immunogenic peptide
7 Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1
5 10 15 Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala Tyr Gly Thr
20 25 30 8 31 PRT artificial sequence anti-HCV immunogenic peptide
8 Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1
5 10 15 Ala Thr Leu Ser Phe Gly Ala Tyr Met Ser Lys Ala His Gly Thr
20 25 30 9 31 PRT artificial sequence anti-HCV immunogenic peptide
9 Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1
5 10 15 Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala Tyr Gly Ile
20 25 30 10 31 PRT artificial sequence anti-HCV immunogenic peptide
10 Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Thr
1 5 10 15 Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly
Val 20 25 30 11 31 PRT artificial sequence anti-HCV immunogenic
peptide 11 Tyr Ala Gly Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser
Val Ala 1 5 10 15 Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala
His Gly Ile 20 25 30 12 31 PRT artificial sequence anti-HCV
immunogenic peptide 12 Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu
Asn Pro Ser Val Ala 1 5 10 15 Ala Thr Leu Ser Phe Gly Ala Tyr Met
Ser Lys Ala His Gly Ile 20 25 30 13 31 PRT artificial sequence
anti-HCV immunogenic peptide 13 Tyr Ala Thr Gln Gly Tyr Lys Val Leu
Val Leu Asn Pro Ser Val Ala 1 5 10 15 Ala Thr Leu Gly Phe Gly Ala
Tyr Met Ser Lys Ala His Gly Thr 20 25 30 14 31 PRT artificial
sequence anti-HCV immunogenic peptide 14 Tyr Ala Ala Gln Gly Tyr
Lys Val Leu Val Leu Asn Pro Ser Val Ala 1 5 10 15 Ala Thr Leu Ser
Phe Gly Ala Tyr Met Ser Lys Ala His Gly Val 20 25 30 15 31 PRT
artificial sequence anti-HCV immunogenic peptide 15 Tyr Ala Thr Gln
Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1 5 10 15 Ala Thr
Leu Ser Phe Gly Ala Tyr Met Ser Lys Ala Tyr Gly Met 20 25 30 16 31
PRT artificial sequence anti-HCV immunogenic peptide 16 Tyr Ala Ala
Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1 5 10 15 Ala
Thr Leu Ser Phe Gly Ala Tyr Met Ser Lys Ala Tyr Gly Val 20 25 30 17
31 PRT artificial sequence anti-HCV immunogenic peptide 17 Tyr Ala
Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1 5 10 15
Ala Thr Leu Gly Phe Gly Thr Tyr Met Ser Lys Ala Tyr Gly Thr 20 25
30 18 31 PRT artificial sequence anti-HCV immunogenic peptide 18
His Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 1 5
10 15 Ala Thr Leu Gly Phe Gly Val Tyr Met Ser Lys Ala Tyr Gly Ile
20 25 30 19 46 PRT artificial sequence anti-HCV immunogenic peptide
19 Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Arg
1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe Gly Ala
Tyr Met 20 25 30 Ser Lys Ala His Gly Ile Glu Pro Asn Ile Arg Thr
Gly Val 35 40 45 20 46 PRT artificial sequence anti-HCV immunogenic
peptide 20 Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys
Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe
Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly Ile Asp Pro Asn Ile
Arg Thr Gly Val 35 40 45 21 46 PRT artificial sequence anti-HCV
immunogenic peptide 21 Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln
Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr
Leu Gly Phe Gly Ala Tyr Met 20 25 30 Ser Lys Ala Tyr Gly Val Asp
Pro Asn Val Arg Thr Gly Val 35 40 45 22 46 PRT artificial sequence
anti-HCV immunogenic peptide 22 Ser Thr Lys Val Pro Ala Ala Tyr Ala
Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala
Ala Thr Leu Gly Phe Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly
Thr Asp Pro Asn Ile Arg Thr Gly Val 35 40 45 23 46 PRT artificial
sequence anti-HCV immunogenic peptide 23 Ser Thr Lys Val Pro Ala
Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro
Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met 20 25 30 Ser Lys
Ala Tyr Gly Thr Asp Pro Asn Ile Arg Thr Gly Val 35 40 45 24 46 PRT
Artificial Sequence anti-HCV immunogenic peptide 24 Ser Thr Lys Val
Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu
Asn Pro Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met 20 25 30
Ser Lys Ala His Gly Ile Glu Pro Asn Ile Arg Thr Gly Val 35 40 45 25
46 PRT artificial sequence anti-HCV immunogenic peptide 25 Ser Thr
Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5 10 15
Val Leu Asn Pro Ser Val Ala Ala Thr Leu Ser Phe Gly Ala Tyr Met 20
25 30 Ser Lys Ala His Gly Thr Asp Pro Asn Ile Arg Thr Gly Val 35 40
45 26 46 PRT artificial sequence anti-HCV immunogenic peptide 26
Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5
10 15 Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr
Met 20 25 30 Ser Lys Ala Tyr Gly Ile Asp Pro Asn Val Arg Thr Gly
Val 35 40 45 27 46 PRT artificial sequence anti-HCV immunogenic
peptide 27 Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys
Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe
Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly Thr Glu Pro Asn Ile
Arg Thr Gly Val 35 40 45 28 46 PRT artificial sequence anti-HCV
immunogenic peptide 28 Ser Asn Lys Val Pro Val Glu Tyr Ala Ala Gln
Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr
Leu Gly Phe Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly Val Asp
Pro Asn Ile Arg Thr Gly Val 35 40 45 29 46 PRT artificial sequence
anti-HCV immunogenic peptide 29 Ser Thr Lys Val Pro Ala Ala Tyr Ala
Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Thr
Ala Thr Leu Gly Phe Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly
Val Asp Pro Asn Ile Arg Thr Gly Val 35 40 45 30 46 PRT artificial
sequence anti-HCV immunogenic peptide 30 Ser Thr Lys Val Pro Ala
Ala Tyr Ala Gly Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro
Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met 20 25 30 Ser Lys
Ala His Gly Ile Asp Pro Asn Ile Arg Thr Gly Val 35 40 45 31 46 PRT
artificial sequence anti-HCV immunogenic peptide 31 Ser Thr Lys Val
Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu
Asn Pro Ser Val Ala Ala Thr Leu Ser Phe Gly Ala Tyr Met 20 25 30
Ser Lys Ala His Gly Ile Asp Pro Asn Ile Arg Thr Gly Val 35 40 45 32
46 PRT artificial sequence anti-HCV immunogenic peptide 32 Ser Thr
Arg Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5 10 15
Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met 20
25 30 Ser Lys Ala His Gly Ile Asp Pro Asn Leu Arg Thr Gly Val 35 40
45 33 46 PRT artificial sequence anti-HCV immunogenic peptide 33
Ser Thr Lys Val Pro Ala Ala Tyr Ala Thr Gln Gly Tyr Lys Val Leu 1 5
10 15 Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr
Met 20 25 30 Ser Lys Ala His Gly Thr Asp Pro Asn Ile Arg Thr Gly
Val 35 40 45 34 46 PRT artificial sequence anti-HCV immunogenic
peptide 34 Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys
Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe
Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly Ile Asp Pro Asn Val
Arg Thr Gly Val 35 40 45 35 46 PRT artificial sequence anti-HCV
immunogenic peptide 35 Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln
Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr
Leu Ser Phe Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly Val Asp
Pro Ser Ile Arg Thr Gly Val 35 40 45 36 46 PRT artificial sequence
anti-HCV immunogenic peptide 36 Ser Thr Lys Val Pro Ala Ala Tyr Ala
Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala
Ala Thr Leu Gly Phe Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly
Val Asp Pro Asn Ile Ser Thr Gly Val 35 40 45 37 46 PRT artificial
sequence anti-HCV immunogenic peptide 37 Ser Thr Lys Val Pro Ala
Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro
Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met 20 25 30 Ser Lys
Ala Tyr Gly Thr Asp Pro Asn Val Arg Thr Gly Val 35 40 45 38 46 PRT
artificial sequence anti-HCV immunogenic peptide 38 Ser Thr Lys Val
Pro Ala Ala Tyr Ala Thr Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu
Asn Pro Ser Val Ala Ala Thr Leu Ser Phe Gly Ala Tyr Met 20 25 30
Ser Lys Ala Tyr Gly Met Asp Pro Asn Leu Arg Thr Gly Val 35 40 45 39
46 PRT Artificial Sequence anti-HCV immunogenic peptide 39 Ser Thr
Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5 10 15
Val Leu Asn Pro Ser Val Ala Ala Thr Leu Ser Phe Gly Ala Tyr Met 20
25 30 Ser Lys Ala His Gly Val Asp Pro Asn Ile Arg Thr Gly Val 35 40
45 40 46 PRT artificial sequence anti-HCV immunogenic peptide 40
Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu 1 5
10 15 Val Leu Asn Pro Ser Val Ala Ala Thr Leu Ser Phe Gly Ala Tyr
Met 20 25 30 Ser Lys Ala Tyr Gly Val Asp Pro Asn Ile Arg Thr Gly
Val 35 40 45 41 46 PRT artificial sequence anti-HCV immunogenic
peptide 41 Ser Thr Arg Val Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys
Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe
Gly Ala Tyr Met 20 25 30 Ser Lys Ala His Gly Thr Asp Pro Asn Ile
Arg Thr Gly Val 35 40 45 42 46 PRT artificial sequence anti-HCV
immunogenic peptide 42 Ser Thr Arg Val Pro Ala Ala Tyr Ala Ala Gln
Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala Ala Thr
Leu Ser Phe Gly Ala Tyr Met 20 25 30 Ser Lys Ala Tyr Gly Val Asp
Pro Asn Ile Arg Thr Gly Val 35 40 45 43 46 PRT artificial sequence
anti-HCV immunogenic peptide 43 Ser Thr Arg Val Pro Ala Ala Tyr Ala
Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro Ser Val Ala
Ala Thr Leu Gly Phe Gly Thr Tyr Met 20 25 30 Ser Lys Ala Tyr Gly
Thr Asp Pro Asn Ile Arg Thr Gly Val 35 40 45 44 46 PRT artificial
sequence anti-HCV immunogenic peptide 44 Ser Thr Lys Val Pro Ala
Ala His Ala Ala Gln Gly Tyr Lys Val Leu 1 5 10 15 Val Leu Asn Pro
Ser Val Ala Ala Thr Leu Gly Phe Gly Val Tyr Met 20 25 30 Ser Lys
Ala Tyr Gly Ile Asp Pro Asn Ile Arg Ser Gly Val 35 40 45 45 45 PRT
artificial sequence anti-HCV immunogenic peptide MISC_FEATURE
(2)..(2) Xaa = L or V MISC_FEATURE (3)..(3) Xaa = L or F
MISC_FEATURE (4)..(4) Xaa = G or S MISC_FEATURE (5)..(5) Xaa = C or
T MISC_FEATURE (6)..(6) Xaa = I or V MISC_FEATURE (7)..(7) Xaa = I
or V MISC_FEATURE (15)..(15) Xaa = K, R or T MISC_FEATURE
(17)..(17) Xaa = Q or E MISC_FEATURE (18)..(18) Xaa = V or N
MISC_FEATURE (19)..(19) Xaa = D, E or C MISC_FEATURE (22)..(22) Xaa
= V or A MISC_FEATURE (24)..(24) Xaa = V, I, E or M MISC_FEATURE
(25)..(25) Xaa = L or V MISC_FEATURE (29)..(29) Xaa = T, K or A
MISC_FEATURE (30)..(30) Xaa = Q or H MISC_FEATURE (31)..(31) Xaa =
S or T MISC_FEATURE (34)..(34) Xaa = A or G MISC_FEATURE (35)..(35)
Xaa = T or S MISC_FEATURE (36)..(36) Xaa = C or A MISC_FEATURE
(37)..(37) Xaa = V, I or T MISC_FEATURE (40)..(40) Xaa = V or A
MISC_FEATURE (41)..(41) Xaa = C or M MISC_FEATURE (45)..(45) Xaa =
Y or F 45 Gly Xaa Xaa Xaa Xaa Xaa Xaa Thr Ser Leu Thr Gly Arg Asp
Xaa Asn 1 5 10
15 Xaa Xaa Xaa Gly Glu Xaa Gln Xaa Xaa Ser Thr Ala Xaa Xaa Xaa Phe
20 25 30 Leu Xaa Xaa Xaa Xaa Asn Gly Xaa Xaa Trp Thr Val Xaa 35 40
45 46 63 PRT artificial sequence anti-HCV immunogenic peptide
MISC_FEATURE (2)..(2) Xaa = P, S, or H MISC_FEATURE (5)..(5) Xaa =
A or T MISC_FEATURE (7)..(7) Xaa = S, A, T or C MISC_FEATURE
(8)..(8) Xaa = Q or R MISC_FEATURE (13)..(13) Xaa = L or V
MISC_FEATURE (14)..(14) Xaa = L or F MISC_FEATURE (15)..(15) Xaa =
G or S MISC_FEATURE (16)..(16) Xaa = C or T MISC_FEATURE (17)..(17)
Xaa = I or V MISC_FEATURE (18)..(18) Xaa = I or V MISC_FEATURE
(26)..(26) Xaa = K, R or T MISC_FEATURE (28)..(28) Xaa = Q or E
MISC_FEATURE (29)..(29) Xaa = V or N MISC_FEATURE (30)..(30) Xaa =
D, E or C MISC_FEATURE (33)..(33) Xaa = V or A MISC_FEATURE
(35)..(35) Xaa = V, I, E or M MISC_FEATURE (36)..(36) Xaa = L or V
MISC_FEATURE (40)..(40) Xaa = T, K or A MISC_FEATURE (41)..(41) Xaa
= Q or H MISC_FEATURE (42)..(42) Xaa = S or T MISC_FEATURE
(45)..(45) Xaa = A or G MISC_FEATURE (46)..(46) Xaa = T or S
MISC_FEATURE (47)..(47) Xaa = C or A MISC_FEATURE (48)..(48) Xaa =
V, I or T MISC_FEATURE (51)..(51) Xaa = V or A MISC_FEATURE
(52)..(52) Xaa = C or M MISC_FEATURE (56)..(56) Xaa = Y or F
MISC_FEATURE (61)..(61) Xaa = S, T or A MISC_FEATURE (62)..(62) Xaa
= K or R MISC_FEATURE (63)..(63) Xaa = T or I 46 Ala Xaa Ile Thr
Xaa Tyr Xaa Xaa Gln Thr Arg Gly Xaa Xaa Xaa Xaa 1 5 10 15 Xaa Xaa
Thr Ser Leu Thr Gly Arg Asp Xaa Asn Xaa Xaa Xaa Gly Glu 20 25 30
Xaa Gln Xaa Xaa Ser Thr Ala Xaa Xaa Xaa Phe Leu Xaa Xaa Xaa Xaa 35
40 45 Asn Gly Xaa Xaa Trp Thr Val Xaa His Gly Ala Gly Xaa Xaa Xaa
50 55 60 47 45 PRT artificial sequence anti-HCV immunogenic peptide
47 Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn
1 5 10 15 Gln Val Asp Gly Glu Val Gln Val Leu Ser Thr Ala Thr Gln
Ser Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val
Tyr 35 40 45 48 45 PRT artificial sequence anti-HCV immunogenic
peptide 48 Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn 1 5 10 15 Gln Val Glu Gly Glu Val Gln Ile Val Ser Thr Ala
Ala Gln Thr Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp
Thr Val Tyr 35 40 45 49 45 PRT artificial sequence anti-HCV
immunogenic peptide 49 Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr
Gly Arg Asp Lys Asn 1 5 10 15 Gln Val Glu Gly Glu Val Gln Ile Val
Ser Thr Ala Thr Gln Thr Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly
Val Cys Trp Thr Val Tyr 35 40 45 50 45 PRT artificial sequence
anti-HCV immunogenic peptide 50 Gly Leu Leu Gly Cys Ile Ile Thr Ser
Leu Thr Gly Arg Asp Lys Asn 1 5 10 15 Gln Val Glu Gly Glu Val Gln
Val Val Ser Thr Ala Thr Gln Ser Phe 20 25 30 Leu Ala Thr Cys Val
Asn Gly Val Cys Trp Thr Val Tyr 35 40 45 51 45 PRT artificial
sequence anti-HCV immunogenic peptide 51 Gly Leu Leu Gly Cys Ile
Ile Thr Ser Leu Thr Gly Arg Asp Arg Asn 1 5 10 15 Gln Val Glu Gly
Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe 20 25 30 Leu Ala
Thr Cys Val Asn Gly Val Cys Trp Thr Val Tyr 35 40 45 52 45 PRT
artificial sequence anti-HCV immunogenic peptide 52 Gly Leu Leu Gly
Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5 10 15 Gln Val
Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe 20 25 30
Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr 35 40 45 53 45
PRT artificial sequence anti-HCV immunogenic peptide 53 Gly Leu Phe
Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5 10 15 Gln
Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe 20 25
30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Tyr 35 40 45 54
45 PRT artificial sequence anti-HCV immunogenic peptide 54 Gly Val
Leu Gly Cys Val Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5 10 15
Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe 20
25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr 35 40 45
55 45 PRT artificial sequence anti-HCV immunogenic peptide 55 Gly
Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5 10
15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe
20 25 30 Leu Ala Thr Cys Val Asn Gly Ala Cys Trp Thr Val Phe 35 40
45 56 45 PRT artificial sequence anti-HCV immunogenic peptide 56
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Phe 35
40 45 57 45 PRT artificial sequence anti-HCV immunogenic peptide 57
Gly Val Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 58 45 PRT artificial sequence anti-HCV immunogenic peptide 58
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Phe 35
40 45 59 45 PRT artificial sequence anti-HCV immunogenic peptide 59
Gly Leu Leu Gly Cys Ile Val Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Ala Cys Trp Thr Val Phe 35
40 45 60 45 PRT artificial sequence anti-HCV immunogenic peptide 60
Gly Val Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Phe 35
40 45 61 45 PRT artificial sequence anti-HCV immunogenic peptide 61
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Asp Gly Glu Val Gln Val Leu Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 62 45 PRT artificial sequence anti-HCV immunogenic peptide 62
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Ala Cys Trp Thr Val Tyr 35
40 45 63 45 PRT artificial sequence anti-HCV immunogenic peptide 63
Gly Val Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr His Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 64 45 PRT artificial sequence anti-HCV immunogenic peptide 64
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Ala Cys Trp Thr Val Tyr 35
40 45 65 45 PRT artificial sequence anti-HCV immunogenic peptide 65
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Ser Cys Val Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 66 45 PRT artificial sequence anti-HCV immunogenic peptide 66
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Thr Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 67 45 PRT artificial sequence anti-HCV immunogenic peptide 67
Gly Val Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 68 45 PRT artificial sequence anti-HCV immunogenic peptide 68
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Lys Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 69 45 PRT artificial sequence anti-HCV immunogenic peptide 69
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Lys Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Ala Cys Trp Thr Val Tyr 35
40 45 70 45 PRT artificial sequence anti-HCV immunogenic peptide 70
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Thr
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Phe 35
40 45 71 45 PRT artificial sequence anti-HCV immunogenic peptide 71
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Glu Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 72 45 PRT artificial sequence anti-HCV immunogenic peptide 72
Gly Leu Phe Gly Cys Ile Val Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Ala Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 73 45 PRT artificial sequence anti-HCV immunogenic peptide 73
Gly Leu Phe Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 74 45 PRT artificial sequence anti-HCV immunogenic peptide 74
Gly Leu Phe Gly Cys Ile Val Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Ala Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Ala Cys Trp Thr Val Tyr 35
40 45 75 45 PRT artificial sequence anti-HCV immunogenic peptide 75
Gly Leu Leu Gly Cys Ile Val Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Phe 35
40 45 76 45 PRT artificial sequence anti-HCV immunogenic peptide 76
Gly Leu Phe Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Arg Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 77 45 PRT artificial sequence anti-HCV immunogenic peptide 77
Gly Leu Phe Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 78 45 PRT artificial sequence anti-HCV immunogenic peptide 78
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Arg Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Phe 35
40 45 79 45 PRT artificial sequence anti-HCV immunogenic peptide 79
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Arg Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Met Val Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val Tyr 35
40 45 80 45 PRT artificial sequence anti-HCV immunogenic peptide 80
Gly Leu Phe Ser Thr Ile Ile Thr Ser Leu Thr Gly Arg Asp Thr Asn 1 5
10 15 Glu Asn Cys Gly Glu Val Gln Val Leu Ser Thr Ala Thr Gln Ser
Phe 20 25 30 Leu Gly Thr Ala Val Asn Gly Val Met Trp Thr Val Tyr 35
40 45 81 63 PRT artificial sequence anti-HCV immunogenic peptide 81
Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys 1 5
10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Asp Gly
Glu 20 25 30 Val Gln Val Leu Ser Thr Ala Thr Gln Ser Phe Leu Ala
Thr Cys Val 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala
Gly Ser Lys Thr 50 55 60 82 63 PRT artificial sequence anti-HCV
immunogenic peptide 82 Ala Pro Ile Thr Ala Tyr Ala Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Ile Val Ser Thr Ala
Ala Gln Thr Phe Leu Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp
Thr Val Tyr His Gly Ala Gly Thr Arg Thr 50 55 60 83 63 PRT
artificial sequence anti-HCV immunogenic peptide 83 Ala Pro Ile Thr
Ala Tyr Thr Gln Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile
Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30
Val Gln Ile Val Ser Thr Ala Thr Gln Thr Phe Leu Ala Thr Cys Ile 35
40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Thr Arg Thr
50 55 60 84 63 PRT artificial sequence anti-HCV immunogenic peptide
84 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys
1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu
Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu
Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly
Ala Gly Ser Lys Thr 50 55 60 85 63 PRT artificial sequence anti-HCV
immunogenic peptide 85 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Arg Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr
Cys Val 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly
Ser Lys Thr 50 55 60 86 63 PRT artificial sequence anti-HCV
immunogenic peptide 86 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp
Thr Val Tyr His Gly Ala Gly Ser Lys Thr 50 55 60 87 63 PRT
artificial sequence anti-HCV immunogenic peptide 87 Ala Pro Ile Thr
Ala Tyr Ser Gln Gln Thr Arg Gly Leu Phe Gly Cys 1 5 10 15 Ile Ile
Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30
Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Val 35
40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys Thr
50 55 60 88 63 PRT artificial sequence anti-HCV immunogenic peptide
88 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Val Leu Gly Cys
1 5 10 15 Val Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu
Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu
Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly
Ala Gly Ser Lys Thr 50 55 60 89 63 PRT artificial sequence anti-HCV
immunogenic peptide 89 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Ala Cys Trp
Thr Val Phe His Gly Ala Gly Ser Lys Thr 50 55 60 90 63 PRT
artificial sequence anti-HCV immunogenic peptide 90 Ala Pro Ile Thr
Ala Tyr Ser Arg Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile
Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30
Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Val 35
40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys Thr
50 55 60 91 63 PRT artificial sequence anti-HCV immunogenic peptide
91 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys
1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu
Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu
Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp Thr Val Phe His Gly
Ala Gly Ala Lys Thr 50 55 60 92 63 PRT artificial sequence anti-HCV
immunogenic peptide 92 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp
Thr Val Phe His Gly Ala Gly Ser Lys Thr 50 55 60 93 63 PRT
artificial sequence anti-HCV immunogenic peptide 93 Ala Pro Ile Thr
Ala Tyr Ser Gln Gln Thr Arg Gly Val Leu Gly Cys 1 5 10 15 Ile Ile
Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30
Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Ile 35
40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys Thr
50 55 60 94 63 PRT artificial sequence anti-HCV immunogenic peptide
94 Ala Pro Ile Thr Ala Tyr Ala Gln Gln Thr Arg Gly Leu Leu Gly Cys
1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu
Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu
Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp Thr Val Phe His Gly
Ala Gly Ser Lys Thr 50 55 60 95 63 PRT artificial sequence anti-HCV
immunogenic peptide 95 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Val Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Ala Cys Trp
Thr Val Phe His Gly Ala Gly Ser Lys Thr 50 55 60 96 63 PRT
artificial sequence anti-HCV immunogenic peptide 96 Ala Pro Ile Thr
Ala Tyr Ala Gln Gln Thr Arg Gly Val Leu Gly Cys 1 5 10 15 Ile Ile
Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30
Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Val 35
40 45 Asn Gly Val Cys Trp Thr Val Phe His Gly Ala Gly Ser Lys Thr
50 55 60 97 63 PRT artificial sequence anti-HCV immunogenic peptide
97 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys
1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Asp
Gly Glu 20 25 30 Val Gln Val Leu Ser Thr Ala Thr Gln Ser Phe Leu
Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly
Ala Gly Ser Lys Thr 50 55 60 98 63 PRT artificial sequence anti-HCV
immunogenic peptide 98 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Ala Cys Trp
Thr Val Tyr His Gly Ala Gly Ser Lys Thr 50 55 60 99 63 PRT
artificial sequence anti-HCV immunogenic peptide 99 Ala Pro Ile Thr
Ala Tyr Ser Gln Gln Thr Arg Gly Val Leu Gly Cys 1 5 10 15 Ile Ile
Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30
Val Gln Val Val Ser Thr Ala Thr His Ser Phe Leu Ala Thr Cys Ile 35
40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys Thr
50 55 60 100 63 PRT artificial sequence anti-HCV immunogenic
peptide 100 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu
Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln
Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe Leu Ala Thr Cys Ile 35 40 45 Asn Gly Ala Cys Trp Thr Val Tyr
His Gly Ala Gly Ser Lys Thr 50 55 60 101 63 PRT artificial sequence
anti-HCV immunogenic peptide 101 Ala Pro Ile Thr Ala Tyr Ser Gln
Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr
Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val
Ser Thr Ala Thr Gln Ser Phe Leu Ala Ser Cys Val 35 40 45 Asn Gly
Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys Thr 50 55 60 102 63
PRT artificial sequence anti-HCV immunogenic peptide 102 Ala Pro
Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15
Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20
25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys
Thr 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser
Lys Thr 50 55 60 103 63 PRT artificial sequence anti-HCV
immunogenic peptide 103 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Val Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp
Thr Val Tyr His Gly Ala Gly Ser Lys Thr 50 55 60 104 63 PRT
artificial sequence anti-HCV immunogenic peptide 104 Ala Pro Ile
Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile
Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25
30 Val Gln Val Val Ser Thr Ala Thr Lys Ser Phe Leu Ala Thr Cys Val
35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys
Thr 50 55 60 105 63 PRT artificial sequence anti-HCV immunogenic
peptide 105 Ala Pro Ile Thr Ala Tyr Cys Gln Gln Thr Arg Gly Leu Leu
Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln
Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr
His Gly Ala Gly Ser Lys Thr 50 55 60 106 63 PRT artificial sequence
anti-HCV immunogenic peptide 106 Ala Pro Ile Thr Ala Tyr Ser Gln
Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr
Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val
Ser Thr Ala Lys Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly
Ala Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys Thr 50 55 60 107 63
PRT artificial sequence anti-HCV immunogenic peptide 107 Ala Pro
Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15
Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20
25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys
Val 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Thr
Lys Thr 50 55 60 108 63 PRT artificial sequence anti-HCV
immunogenic peptide 108 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Thr Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp
Thr Val Phe His Gly Ala Gly Ser Lys Thr 50 55 60 109 63 PRT
artificial sequence anti-HCV immunogenic peptide 109 Ala Pro Ile
Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile
Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25
30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Val
35 40 45 Asn Gly Ala Cys Trp Thr Val Tyr His Gly Ala Gly Thr Lys
Thr 50 55 60 110 63 PRT artificial sequence anti-HCV immunogenic
peptide 110 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu
Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln
Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp Thr Val Phe
His Gly Ala Gly Thr Lys Thr 50 55 60 111 63 PRT artificial sequence
anti-HCV immunogenic peptide 111 Ala Pro Ile Thr Ala Tyr Ser Gln
Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr
Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Glu Val
Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly
Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys Ile 50 55 60 112 63
PRT artificial sequence anti-HCV immunogenic peptide 112 Ala Pro
Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Phe Gly Cys 1 5 10 15
Ile Val Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20
25 30 Ala Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys
Val 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser
Lys Thr 50 55 60 113 63 PRT artificial sequence anti-HCV
immunogenic peptide 113 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Phe Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp
Thr Val Tyr His Gly Ala Gly Ser Lys Thr 50 55 60 114 63 PRT
artificial sequence anti-HCV immunogenic peptide 114 Ala Pro Ile
Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Ala Gly Cys 1 5 10 15 Ile
Val Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25
30 Ala Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Val
35 40 45 Asn Gly Ala Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys
Thr 50 55 60 115 63 PRT artificial sequence anti-HCV immunogenic
peptide 115 Ala Pro Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Val Leu
Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln
Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp Thr Val Phe
His Gly Ala Gly Ser Lys Thr 50 55 60 116 63 PRT artificial sequence
anti-HCV immunogenic peptide 116 Ala Pro Ile Thr Thr Tyr Ser Gln
Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr
Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val
Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly
Val Cys Trp Thr Val Tyr His Gly Ala Gly Thr Lys Thr 50 55 60 117 63
PRT artificial sequence anti-HCV immunogenic peptide 117 Ala Ser
Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Val Leu Gly Cys 1 5 10 15
Ile Val Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20
25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys
Ile 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser
Lys Thr 50 55 60 118 63 PRT artificial sequence anti-HCV
immunogenic peptide 118 Ala Ser Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Val Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp
Thr Val Phe His Gly Ala Gly Ser Lys Thr 50 55 60 119 63 PRT
artificial sequence anti-HCV immunogenic peptide 119 Ala Pro Ile
Thr Ala Tyr Ala Gln Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile
Val Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu Gly Glu 20 25
30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Val
35 40 45 Asn Gly Val Cys Trp Thr Val Phe His Gly Ala Gly Ser Lys
Thr 50 55 60 120 63 PRT artificial sequence anti-HCV
immunogenic peptide 120 Ala Pro Ile Thr Ala Tyr Ala Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Lys Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp
Thr Val Tyr His Gly Ala Gly Ser Lys Thr 50 55 60 121 63 PRT
artificial sequence anti-HCV immunogenic peptide 121 Ala Pro Ile
Thr Ala Tyr Ala Gln Gln Thr Arg Gly Leu Phe Gly Cys 1 5 10 15 Ile
Ile Thr Ser Leu Thr Gly Arg Asp Arg Asn Gln Val Glu Gly Glu 20 25
30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Ile
35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ser Lys
Thr 50 55 60 122 63 PRT artificial sequence anti-HCV immunogenic
peptide 122 Ala Pro Ile Thr Ala Tyr Ala Gln Gln Thr Arg Gly Leu Phe
Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln
Val Glu Gly Glu 20 25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser
Phe Leu Ala Thr Cys Ile 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr
His Gly Ala Gly Ser Lys Thr 50 55 60 123 63 PRT artificial sequence
anti-HCV immunogenic peptide 123 Ala Pro Ile Thr Ala Tyr Ser Gln
Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr
Gly Arg Asp Arg Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Val Val
Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys Ile 35 40 45 Asn Gly
Val Cys Trp Thr Val Phe His Gly Ala Gly Ser Lys Thr 50 55 60 124 63
PRT artificial sequence anti-HCV immunogenic peptide 124 Ala Pro
Ile Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys 1 5 10 15
Ile Ile Thr Ser Leu Thr Gly Arg Asp Arg Asn Gln Val Glu Gly Glu 20
25 30 Val Gln Val Val Ser Thr Ala Thr Gln Ser Phe Leu Ala Thr Cys
Val 35 40 45 Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Ala
Lys Thr 50 55 60 125 63 PRT artificial sequence anti-HCV
immunogenic peptide 125 Ala His Ile Thr Ala Tyr Ser Gln Gln Thr Arg
Gly Leu Leu Gly Cys 1 5 10 15 Ile Ile Thr Ser Leu Thr Gly Arg Asp
Arg Asn Gln Val Glu Gly Glu 20 25 30 Val Gln Met Val Ser Thr Ala
Thr Gln Ser Phe Leu Ala Thr Cys Val 35 40 45 Asn Gly Val Cys Trp
Thr Val Tyr His Gly Ala Gly Ser Lys Thr 50 55 60 126 63 PRT
artificial sequence anti-HCV immunogenic peptide 126 Ala Pro Ile
Thr Ala Tyr Ser Gln Gln Thr Arg Gly Leu Phe Ser Thr 1 5 10 15 Ile
Ile Thr Ser Leu Thr Gly Arg Asp Thr Asn Glu Asn Cys Gly Glu 20 25
30 Val Gln Val Leu Ser Thr Ala Thr Gln Ser Phe Leu Gly Thr Cys Val
35 40 45 Asn Gly Val Met Trp Thr Val Tyr His Gly Ala Gly Ala Lys
Thr 50 55 60 127 32 PRT artificial sequence anti-HCV immunogenic
peptide MISC_FEATURE (2)..(2) Xaa = L or P MISC_FEATURE (4)..(4)
Xaa = A or S MISC_FEATURE (7)..(7) Xaa = A or S MISC_FEATURE
(8)..(8) Xaa = A or S MISC_FEATURE (9)..(9) Xaa = I or V
MISC_FEATURE (14)..(14) Xaa = T, S or A MISC_FEATURE (15)..(15) Xaa
= T or I MISC_FEATURE (16)..(16) Xaa = Q, S or G MISC_FEATURE
(17)..(17) Xaa = N, Q, H, S, Y or T MISC_FEATURE (20)..(20) Xaa = L
or M MISC_FEATURE (24)..(24) Xaa = L or W MISC_FEATURE (30)..(30)
Xaa = A or S MISC_FEATURE (32)..(32) Xaa = L, P or I 127 Ser Xaa
Met Xaa Phe Thr Xaa Xaa Xaa Thr Ser Pro Leu Xaa Xaa Xaa 1 5 10 15
Xaa Thr Leu Xaa Phe Asn Ile Xaa Gly Gly Trp Val Ala Xaa Gln Xaa 20
25 30 128 57 PRT artificial sequence anti-HCV immunogenic peptide
MISC_FEATURE (4)..(4) Xaa = T or S MISC_FEATURE (6)..(6) Xaa = I, T
or V MISC_FEATURE (11)..(11) Xaa = G or A MISC_FEATURE (19)..(19)
Xaa = P or L MISC_FEATURE (21)..(21) Xaa = I or M MISC_FEATURE
(22)..(22) Xaa = A, V or R MISC_FEATURE (24)..(24) Xaa = L or P
MISC_FEATURE (26)..(26) Xaa = A or S MISC_FEATURE (29)..(29) Xaa =
A or S MISC_FEATURE (30)..(30) Xaa = A or S MISC_FEATURE (31)..(31)
Xaa = I or V MISC_FEATURE (36)..(36) Xaa = T, S or A MISC_FEATURE
(37)..(37) Xaa = T or I MISC_FEATURE (38)..(38) Xaa = Q, S or G
MISC_FEATURE (39)..(39) Xaa = N, Q, H, S, Y or T MISC_FEATURE
(42)..(42) Xaa = L or M MISC_FEATURE (46)..(46) Xaa = L or W
MISC_FEATURE (52)..(52) Xaa = A or S MISC_FEATURE (54)..(54) Xaa =
L, P or I MISC_FEATURE (55)..(55) Xaa = A or R MISC_FEATURE
(56)..(56) Xaa = P, A or D MISC_FEATURE (57)..(57) Xaa = P, A or S
128 Asn Phe Ile Xaa Gly Xaa Gln Tyr Leu Ala Xaa Leu Ser Thr Leu Pro
1 5 10 15 Gly Asn Xaa Ala Xaa Xaa Ser Xaa Met Xaa Phe Thr Xaa Xaa
Xaa Thr 20 25 30 Ser Pro Leu Xaa Xaa Xaa Xaa Thr Leu Xaa Phe Asn
Ile Xaa Gly Gly 35 40 45 Trp Val Ala Xaa Gln Xaa Xaa Xaa Xaa 50 55
129 32 PRT artificial sequence anti-HCV immunogenic peptide 129 Ser
Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu Thr Thr Gln 1 5 10
15 Asn Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln Leu
20 25 30 130 32 PRT artificial sequence anti-HCV immunogenic
peptide 130 Ser Leu Met Ala Phe Thr Ala Ala Val Thr Ser Pro Leu Thr
Thr Ser 1 5 10 15 Gln Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val
Ala Ala Gln Leu 20 25 30 131 32 PRT artificial sequence anti-HCV
immunogenic peptide 131 Ser Leu Met Ala Phe Thr Ala Ala Val Thr Ser
Pro Leu Thr Thr Gly 1 5 10 15 Gln Thr Leu Leu Phe Asn Ile Leu Gly
Gly Trp Val Ala Ala Gln Leu 20 25 30 132 32 PRT artificial sequence
anti-HCV immunogenic peptide 132 Ser Leu Met Ala Phe Thr Ala Ser
Ile Thr Ser Pro Leu Thr Thr Gln 1 5 10 15 His Thr Leu Leu Phe Asn
Ile Leu Gly Gly Trp Val Ala Ala Gln Leu 20 25 30 133 32 PRT
artificial sequence anti-HCV immunogenic peptide 133 Ser Leu Met
Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu Thr Thr Gln 1 5 10 15 Ser
Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln Leu 20 25
30 134 32 PRT artificial sequence anti-HCV immunogenic peptide 134
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu Thr Thr Gln 1 5
10 15 Tyr Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln
Leu 20 25 30 135 32 PRT artificial sequence anti-HCV immunogenic
peptide 135 Ser Leu Met Ala Phe Thr Ala Ser Val Thr Ser Pro Leu Thr
Thr Gln 1 5 10 15 Asn Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val
Ala Ala Gln Leu 20 25 30 136 32 PRT artificial sequence anti-HCV
immunogenic peptide 136 Ser Leu Met Ala Phe Thr Ala Ser Val Thr Ser
Pro Leu Thr Thr Gln 1 5 10 15 Ser Thr Leu Leu Phe Asn Ile Leu Gly
Gly Trp Val Ala Ala Gln Leu 20 25 30 137 32 PRT artificial sequence
anti-HCV immunogenic peptide 137 Ser Leu Met Ala Phe Thr Ala Ser
Ile Thr Ser Pro Leu Thr Thr Gln 1 5 10 15 Thr Thr Leu Met Phe Asn
Ile Leu Gly Gly Trp Val Ala Ala Gln Leu 20 25 30 138 32 PRT
artificial sequence anti-HCV immunogenic peptide 138 Ser Leu Met
Ala Phe Thr Ala Ser Val Thr Ser Pro Leu Thr Thr Gln 1 5 10 15 Tyr
Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln Ile 20 25
30 139 32 PRT artificial sequence anti-HCV immunogenic peptide 139
Ser Pro Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu Thr Thr Gln 1 5
10 15 His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln
Leu 20 25 30 140 32 PRT artificial sequence anti-HCV immunogenic
peptide 140 Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu Thr
Ile Gln 1 5 10 15 His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val
Ala Ala Gln Pro 20 25 30 141 32 PRT artificial sequence anti-HCV
immunogenic peptide 141 Ser Leu Met Ala Phe Thr Ser Ser Ile Thr Ser
Pro Leu Thr Thr Gln 1 5 10 15 Ser Thr Leu Leu Phe Asn Ile Leu Gly
Gly Trp Val Ala Ala Gln Leu 20 25 30 142 32 PRT artificial sequence
anti-HCV immunogenic peptide 142 Ser Leu Met Ala Phe Thr Ala Ser
Ile Thr Ser Pro Leu Ser Thr Gln 1 5 10 15 Asn Thr Leu Leu Phe Asn
Ile Trp Gly Gly Trp Val Ala Ala Gln Leu 20 25 30 143 32 PRT
artificial sequence anti-HCV immunogenic peptide 143 Ser Leu Met
Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu Thr Thr Gln 1 5 10 15 Asn
Thr Leu Met Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln Leu 20 25
30 144 32 PRT artificial sequence anti-HCV immunogenic peptide 144
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu Thr Thr Gln 1 5
10 15 His Thr Leu Met Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln
Leu 20 25 30 145 32 PRT artificial sequence anti-HCV immunogenic
peptide 145 Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu Ala
Thr Gln 1 5 10 15 Tyr Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val
Ala Ala Gln Leu 20 25 30 146 32 PRT artificial sequence anti-HCV
immunogenic peptide 146 Ser Leu Met Ser Phe Thr Ala Ala Val Thr Ser
Pro Leu Thr Thr Gln 1 5 10 15 Gln Thr Leu Leu Phe Asn Ile Leu Gly
Gly Trp Val Ala Ser Gln Ile 20 25 30 147 57 PRT artificial sequence
anti-HCV immunogenic peptide 147 Asn Phe Ile Thr Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Asn Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 148 57 PRT artificial sequence
anti-HCV immunogenic peptide 148 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ala Val Thr 20 25 30 Ser Pro Leu Thr
Thr Ser Gln Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Ala Pro 50 55 149 57 PRT artificial sequence
anti-HCV immunogenic peptide 149 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ala Val Thr 20 25 30 Ser Pro Leu Thr
Thr Gly Gln Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Ala Pro 50 55 150 57 PRT artificial sequence
anti-HCV immunogenic peptide 150 Asn Phe Ile Ser Gly Thr Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ala Val Thr 20 25 30 Ser Pro Leu Thr
Thr Ser Gln Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Ala Pro 50 55 151 57 PRT artificial sequence
anti-HCV immunogenic peptide 151 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 152 57 PRT artificial sequence
anti-HCV immunogenic peptide 152 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Ser Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 153 57 PRT artificial sequence
anti-HCV immunogenic peptide 153 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Asn Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 154 57 PRT artificial sequence
anti-HCV immunogenic peptide 154 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Tyr Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 155 57 PRT artificial sequence
anti-HCV immunogenic peptide 155 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Val
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 156 57 PRT artificial sequence
anti-HCV immunogenic peptide 156 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Val Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Asn Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 157 57 PRT artificial sequence
anti-HCV immunogenic peptide 157 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Val Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Ser Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 158 57 PRT artificial sequence
anti-HCV immunogenic peptide 158 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Thr Thr Leu Met Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 159 57 PRT artificial sequence
anti-HCV immunogenic peptide 159 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Val Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Tyr Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Ile Ala Pro Pro 50 55 160 57 PRT artificial sequence
anti-HCV immunogenic peptide 160 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Arg
Ser Pro Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 161 57 PRT artificial
sequence
anti-HCV immunogenic peptide 161 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Ile Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Pro Ala Pro Pro 50 55 162 57 PRT artificial sequence
anti-HCV immunogenic peptide 162 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Ala Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Asn Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Ala 50 55 163 57 PRT artificial sequence
anti-HCV immunogenic peptide 163 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ser Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Ser Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 164 57 PRT artificial sequence
anti-HCV immunogenic peptide 164 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Ser
Thr Gln Asn Thr Leu Leu Phe Asn Ile Trp Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 165 57 PRT artificial sequence
anti-HCV immunogenic peptide 165 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Asn Thr Leu Met Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 166 57 PRT artificial sequence
anti-HCV immunogenic peptide 166 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Met Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln His Thr Leu Met Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 167 57 PRT artificial sequence
anti-HCV immunogenic peptide 167 Asn Phe Ile Ser Gly Val Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Asn Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 168 57 PRT artificial sequence
anti-HCV immunogenic peptide 168 Asn Phe Ile Ser Gly Val Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 169 57 PRT artificial sequence
anti-HCV immunogenic peptide 169 Asn Phe Ile Ser Gly Val Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Tyr Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 170 57 PRT artificial sequence
anti-HCV immunogenic peptide 170 Asn Phe Ile Ser Gly Val Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Val Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Ser Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 171 57 PRT artificial sequence
anti-HCV immunogenic peptide 171 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Leu Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Thr
Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 172 57 PRT artificial sequence
anti-HCV immunogenic peptide 172 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ala Phe Thr Ala Ser Ile Thr 20 25 30 Ser Pro Leu Ala
Thr Gln Tyr Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ala Gln Leu Ala Pro Pro 50 55 173 57 PRT artificial sequence
anti-HCV immunogenic peptide 173 Asn Phe Ile Ser Gly Ile Gln Tyr
Leu Ala Gly Leu Ser Thr Leu Pro 1 5 10 15 Gly Asn Pro Ala Ile Ala
Ser Leu Met Ser Phe Thr Ala Ala Val Thr 20 25 30 Ser Pro Leu Thr
Thr Gln Gln Thr Leu Leu Phe Asn Ile Leu Gly Gly 35 40 45 Trp Val
Ala Ser Gln Ile Arg Asp Ser 50 55 174 29 PRT artificial sequence
anti-HCV immunogenic peptide MISC_FEATURE (1)..(1) Xaa = R or Q
MISC_FEATURE (3)..(3) Xaa = P or A MISC_FEATURE (6)..(6) Xaa = L or
F MISC_FEATURE (9)..(9) Xaa = F or Y MISC_FEATURE (11)..(11) Xaa =
D or E MISC_FEATURE (14)..(14) Xaa = V or S MISC_FEATURE (20)..(20)
Xaa = M or R MISC_FEATURE (23)..(23) Xaa = Y or H MISC_FEATURE
(24)..(24) Xaa = D or N MISC_FEATURE (26)..(26) Xaa = V or I
MISC_FEATURE (27)..(27) Xaa = S, T or K MISC_FEATURE (28)..(28) Xaa
= T, K, I or N MISC_FEATURE (29)..(29) Xaa = L or T 174 Xaa Lys Xaa
Ala Arg Xaa Ile Val Xaa Pro Xaa Leu Gly Xaa Arg Val 1 5 10 15 Cys
Glu Lys Xaa Ala Leu Xaa Xaa Val Xaa Xaa Xaa Xaa 20 25 175 44 PRT
artificial sequence anti-HCV immunogenic peptide MISC_FEATURE
(4)..(4) Xaa = R or Q MISC_FEATURE (6)..(6) Xaa = P or A
MISC_FEATURE (9)..(9) Xaa = L or F MISC_FEATURE (12)..(12) Xaa = F
or Y MISC_FEATURE (14)..(14) Xaa = D or E MISC_FEATURE (17)..(17)
Xaa = V or S MISC_FEATURE (23)..(23) Xaa = M or R MISC_FEATURE
(26)..(26) Xaa = Y or H MISC_FEATURE (27)..(27) Xaa = D or N
MISC_FEATURE (29)..(29) Xaa = V or I MISC_FEATURE (30)..(30) Xaa =
S, T or K MISC_FEATURE (31)..(31) Xaa = T, K, I or N MISC_FEATURE
(32)..(32) Xaa = L or T MISC_FEATURE (33)..(33) Xaa = P or A
MISC_FEATURE (34)..(34) Xaa = Q, L, H, R, K or P MISC_FEATURE
(35)..(35) Xaa = A, T, V or P MISC_FEATURE (39)..(39) Xaa = P, S or
A MISC_FEATURE (40)..(40) Xaa = S or A MISC_FEATURE (42)..(42) Xaa
= G or R MISC_FEATURE (43)..(43) Xaa = F or C 175 Lys Gly Gly Xaa
Lys Xaa Ala Arg Xaa Ile Val Xaa Pro Xaa Leu Gly 1 5 10 15 Xaa Arg
Val Cys Glu Lys Xaa Ala Leu Xaa Xaa Val Xaa Xaa Xaa Xaa 20 25 30
Xaa Xaa Xaa Val Met Gly Xaa Xaa Tyr Xaa Xaa Gln 35 40 176 29 PRT
artificial sequence anti-HCV immunogenic peptide 176 Arg Lys Pro
Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25 177 29 PRT
artificial sequence anti-HCV immunogenic peptide 177 Arg Lys Pro
Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Lys Leu 20 25 178 29 PRT
artificial sequence anti-HCV immunogenic peptide 178 Arg Lys Pro
Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Thr Lys Leu 20 25 179 29 PRT
artificial sequence anti-HCV immunogenic peptide 179 Gln Lys Pro
Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25 180 29 PRT
artificial sequence anti-HCV immunogenic peptide 180 Arg Lys Ala
Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25 181 29 PRT
artificial sequence anti-HCV immunogenic peptide 181 Arg Lys Pro
Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Ile Leu 20 25 182 29 PRT
artificial sequence anti-HCV immunogenic peptide 182 Arg Lys Pro
Ala Arg Leu Ile Val Phe Pro Glu Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25 183 29 PRT
artificial sequence anti-HCV immunogenic peptide 183 Arg Lys Pro
Ala Arg Phe Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25 184 29 PRT
artificial sequence anti-HCV immunogenic peptide 184 Arg Lys Ala
Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asn Val Val Ser Thr Leu 20 25 185 29 PRT
artificial sequence anti-HCV immunogenic peptide 185 Arg Lys Pro
Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Asn Leu 20 25 186 29 PRT
artificial sequence anti-HCV immunogenic peptide 186 Arg Lys Pro
Ala Arg Leu Ile Val Tyr Pro Asp Leu Gly Val Arg Val 1 5 10 15 Cys
Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25 187 29 PRT
artificial sequence anti-HCV immunogenic peptide 187 Arg Lys Pro
Ala Arg Leu Ile Val Tyr Pro Asp Leu Gly Ser Arg Val 1 5 10 15 Cys
Glu Lys Arg Ala Leu His Asp Val Ile Lys Lys Thr 20 25 188 44 PRT
artificial sequence anti-HCV immunogenic peptide 188 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Ala Val Met Gly Pro Ser Tyr Gly Phe Gln 35 40 189 44 PRT
artificial sequence anti-HCV immunogenic peptide 189 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Lys Leu 20 25
30 Pro Leu Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 190 44 PRT
artificial sequence anti-HCV immunogenic peptide 190 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Lys Leu 20 25
30 Pro Pro Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 191 44 PRT
artificial sequence anti-HCV immunogenic peptide 191 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Thr Lys Leu 20 25
30 Pro Leu Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 192 44 PRT
artificial sequence anti-HCV immunogenic peptide 192 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 193 44 PRT
artificial sequence anti-HCV immunogenic peptide 193 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Ala Val Met Gly Ala Ser Tyr Gly Phe Gln 35 40 194 44 PRT
artificial sequence anti-HCV immunogenic peptide 194 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Val Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 195 44 PRT
artificial sequence anti-HCV immunogenic peptide 195 Lys Gly Gly
Gln Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 196 44 PRT
artificial sequence anti-HCV immunogenic peptide 196 Lys Gly Gly
Arg Lys Ala Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 197 44 PRT
artificial sequence anti-HCV immunogenic peptide 197 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Ile Leu 20 25
30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 198 44 PRT
artificial sequence anti-HCV immunogenic peptide 198 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Glu Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 199 44 PRT
artificial sequence anti-HCV immunogenic peptide 199 Lys Gly Gly
Arg Lys Pro Ala Arg Phe Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Lys Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 200 44 PRT
artificial sequence anti-HCV immunogenic peptide 200 Lys Gly Gly
Arg Lys Pro Ala Arg Phe Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 201 44 PRT
artificial sequence anti-HCV immunogenic peptide 201 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Arg Ala Val Met Gly Ser Ser Tyr Gly Cys Gln 35 40 202 44 PRT
artificial sequence anti-HCV immunogenic peptide 202 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Pro Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 203 44 PRT
artificial sequence anti-HCV immunogenic peptide 203 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro His Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 204 44 PRT
artificial sequence anti-HCV immunogenic peptide 204 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15 Val
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20 25
30 Pro Gln Ala Val Met Gly Ser Ala Tyr Gly Phe Gln 35 40 205 44 PRT
artificial sequence anti-HCV immunogenic peptide 205 Lys Gly Gly
Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1
5 10 15 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr
Leu 20 25 30 Pro Gln Ala Val Met Gly Ser Ser Tyr Arg Phe Gln 35 40
206 44 PRT artificial sequence anti-HCV immunogenic peptide 206 Lys
Gly Gly Arg Lys Ala Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10
15 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu
20 25 30 Pro Gln Ala Val Met Gly Pro Ser Tyr Gly Phe Gln 35 40 207
44 PRT artificial sequence anti-HCV immunogenic peptide 207 Lys Gly
Gly Arg Lys Ala Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15
Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asn Val Val Ser Thr Leu 20
25 30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 208 44
PRT artificial sequence anti-HCV immunogenic peptide 208 Lys Gly
Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15
Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Asn Leu 20
25 30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 209 44
PRT artificial sequence anti-HCV immunogenic peptide 209 Lys Gly
Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Glu Leu Gly 1 5 10 15
Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20
25 30 Pro Gln Ala Val Met Gly Pro Ser Tyr Gly Phe Gln 35 40 210 44
PRT artificial sequence anti-HCV immunogenic peptide 210 Lys Gly
Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1 5 10 15
Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20
25 30 Pro His Thr Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 211 44
PRT artificial sequence anti-HCV immunogenic peptide 211 Lys Gly
Gly Arg Lys Pro Ala Arg Leu Ile Val Tyr Pro Asp Leu Gly 1 5 10 15
Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 20
25 30 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln 35 40 212 44
PRT artificial sequence anti-HCV immunogenic peptide 212 Lys Gly
Gly Arg Lys Pro Ala Arg Leu Ile Val Tyr Pro Asp Leu Gly 1 5 10 15
Ser Arg Val Cys Glu Lys Arg Ala Leu His Asp Val Ile Lys Lys Thr 20
25 30 Ala Leu Ala Val Met Gly Ala Ala Tyr Gly Phe Gln 35 40 213 10
PRT artificial sequence anti-HCV immunogenic peptide MISC_FEATURE
(2)..(2) Xaa = L or V MISC_FEATURE (3)..(3) Xaa = L or F
MISC_FEATURE (4)..(4) Xaa = G or S MISC_FEATURE (5)..(5) Xaa = C or
T MISC_FEATURE (6)..(6) Xaa = I or V MISC_FEATURE (7)..(7) Xaa = I
or V 213 Gly Xaa Xaa Xaa Xaa Xaa Xaa Thr Ser Leu 1 5 10 214 10 PRT
artificial sequence anti-HCV immunogenic peptide 214 Gly Leu Leu
Gly Cys Ile Ile Thr Ser Leu 1 5 10 215 10 PRT artificial sequence
anti-HCV immunogenic peptide 215 Gly Leu Phe Gly Cys Ile Ile Thr
Ser Leu 1 5 10 216 10 PRT artificial sequence anti-HCV immunogenic
peptide 216 Gly Val Leu Gly Cys Val Ile Thr Ser Leu 1 5 10 217 10
PRT artificial sequence anti-HCV immunogenic peptide 217 Gly Val
Leu Gly Cys Ile Ile Thr Ser Leu 1 5 10 218 10 PRT artificial
sequence anti-HCV immunogenic peptide 218 Gly Leu Leu Gly Cys Ile
Val Thr Ser Leu 1 5 10 219 10 PRT artificial sequence anti-HCV
immunogenic peptide 219 Gly Leu Phe Gly Cys Ile Val Thr Ser Leu 1 5
10 220 10 PRT artificial sequence anti-HCV immunogenic peptide 220
Gly Leu Phe Ser Thr Ile Ile Thr Ser Leu 1 5 10 221 9 PRT artificial
sequence anti-HCV immunogenic peptide MISC_FEATURE (4)..(4) Xaa =
T, S or A MISC_FEATURE (5)..(5) Xaa = T or I MISC_FEATURE (6)..(6)
Xaa = Q, S or G MISC_FEATURE (7)..(7) Xaa = N, Q, H, S, Y or T 221
Ser Pro Leu Xaa Xaa Xaa Xaa Thr Leu 1 5 222 9 PRT artificial
sequence anti-HCV immunogenic peptide 222 Ser Pro Leu Thr Thr Gln
Asn Thr Leu 1 5 223 9 PRT artificial sequence anti-HCV immunogenic
peptide 223 Ser Pro Leu Thr Thr Ser Gln Thr Leu 1 5 224 9 PRT
artificial sequence anti-HCV immunogenic peptide 224 Ser Pro Leu
Thr Thr Gly Gln Thr Leu 1 5 225 9 PRT artificial sequence anti-HCV
immunogenic peptide 225 Ser Pro Leu Thr Thr Gln His Thr Leu 1 5 226
9 PRT artificial sequence anti-HCV immunogenic peptide 226 Ser Pro
Leu Thr Thr Gln Ser Thr Leu 1 5 227 9 PRT artificial sequence
anti-HCV immunogenic peptide 227 Ser Pro Leu Thr Thr Gln Tyr Thr
Leu 1 5 228 9 PRT artificial sequence anti-HCV immunogenic peptide
228 Ser Pro Leu Thr Thr Gln Thr Thr Leu 1 5 229 9 PRT artificial
sequence anti-HCV immunogenic peptide 229 Ser Pro Leu Thr Ile Gln
His Thr Leu 1 5 230 9 PRT artificial sequence anti-HCV immunogenic
peptide 230 Ser Pro Leu Ser Thr Gln Asn Thr Leu 1 5 231 9 PRT
artificial sequence anti-HCV immunogenic peptide 231 Ser Pro Leu
Ala Thr Gln Tyr Thr Leu 1 5 232 9 PRT artificial sequence anti-HCV
immunogenic peptide 232 Ser Pro Leu Thr Thr Gln Gln Thr Leu 1 5 233
16 PRT artificial sequence anti-HCV immunogenic peptide 233 Arg Tyr
Met Ile Leu Gly Leu Leu Ala Leu Ala Ala Val Cys Ser Ala 1 5 10 15
234 137 PRT artificial sequence anti-HCV immunogenic peptide 234
Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn 1 5
10 15 Gln Val Glu Gly Glu Val Gln Ile Val Ser Thr Ala Thr Gln Thr
Phe 20 25 30 Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr
Tyr Ala Ala 35 40 45 Gln Gly Tyr Lys Val Arg Val Leu Asn Pro Ser
Val Ala Ala Thr Leu 50 55 60 Gly Phe Gly Ala Tyr Met Ser Lys Ala
His Gly Ile Ser Leu Met Ala 65 70 75 80 Phe Thr Ala Ser Ile Thr Ser
Pro Leu Thr Thr Gln Asn Thr Leu Leu 85 90 95 Phe Asn Ile Leu Gly
Gly Trp Val Ala Ala Gln Leu Arg Lys Pro Ala 100 105 110 Arg Leu Ile
Val Phe Pro Asp Leu Gly Val Arg Val Cys Glu Lys Met 115 120 125 Ala
Leu Tyr Asp Val Val Ser Thr Leu 130 135 235 417 DNA artificial
sequence polyEp-WT coding sequence 235 atgggcctgc ttggctgtat
catcactagc ctcacaggtc gggacaagaa ccaggtcgat 60 ggggaggttc
aggtgctctc caccgcaacg caatctttcc tggcgacctg cgtcaatggc 120
gtgtgttgga ccgtctacta tgcagcccaa gggtacaagg tgcgcgtcct aaacccgtcc
180 gttgccgcca cattgggctt tggagcgtat atgtccaagg cacatggcat
ctcactgatg 240 gcattcacag cctctatcac cagtccgctc accacccaga
ataccctcct attcaacatc 300 ttagggggat gggtggctgc tcaactccgc
aagccagctc gccttatcgt attcccagac 360 ctgggggtac gtgtatgcga
gaagatggcc ctttacgacg tggtctccac cctttag 417 236 417 DNA artificial
sequence polyEp-C coding sequence 236 atgggactgc tgggctgcat
catcaccagc ctcaccggca gagacaagaa tcaggtggac 60 ggcgaggtgc
aggtgctgag caccgccaca cagagcttcc tggccacctg cgtgaacggc 120
gtgtgctgga ccgtgtacta cgccgcccag ggctacaagg tgagagtgct gaaccccagc
180 gtggccgcta ccctgggctt cggcgcctac atgagcaagg cccacggcat
cagcctcatg 240 gccttcaccg ccagcatcac aagccctctc accacccaga
acaccctgct gttcaacatc 300 ctgggcggct gggtggccgc tcagctgaga
aagcccgcca gactcatcgt gttccccgac 360 ctgggcgtga gagtgtgcga
gaagatggcc ctgtacgacg tggtgagcac cctgtga 417 237 465 DNA artificial
sequence polyEp-E3 coding sequence 237 atgaggtaca tgattttagg
cttgctcgcc cttgcggcag tctgcagcgc tggcctgctt 60 ggctgtatca
tcactagcct cacaggtcgg gacaagaacc aggtcgatgg ggaggttcag 120
gtgctctcca ccgcaacgca atctttcctg gcgacctgcg tcaatggcgt gtgttggacc
180 gtctactatg cagcccaagg gtacaaggtg cgcgtcctaa acccgtccgt
tgccgccaca 240 ttgggctttg gagcgtatat gtccaaggca catggcatct
cactgatggc attcacagcc 300 tctatcacca gtccgctcac cacccagaat
accctcctat tcaacatctt agggggatgg 360 gtggctgctc aactccgcaa
gccagctcgc cttatcgtat tcccagacct gggggtacgt 420 gtatgcgaga
agatggccct ttacgacgtg gtctccaccc tttag 465
* * * * *