U.S. patent application number 16/812322 was filed with the patent office on 2020-08-27 for methods of treating and preventing infections.
This patent application is currently assigned to Longhorn Vaccines and Diagnostics, LLC. The applicant listed for this patent is Longhorn Vaccines and Diagnostics, LLC. Invention is credited to Luke T. Daum, Gerald W. Fischer.
Application Number | 20200268874 16/812322 |
Document ID | / |
Family ID | 1000004827959 |
Filed Date | 2020-08-27 |
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United States Patent
Application |
20200268874 |
Kind Code |
A1 |
Fischer; Gerald W. ; et
al. |
August 27, 2020 |
Methods of Treating and Preventing Infections
Abstract
The invention relates to composite antigens comprising a peptide
with contiguous amino acid sequence derived from a plurality of
antigenic epitopes of one or more pathogens that induces an immune
response in a mammal that is protective against infection by the
one or more pathogens. In addition, the invention relates to
vaccines comprising composite antigens and to method for treating
and preventing an infection.
Inventors: |
Fischer; Gerald W.;
(Bethesda, MD) ; Daum; Luke T.; (San Antonio,
TX) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Longhorn Vaccines and Diagnostics, LLC |
Bethesda |
MD |
US |
|
|
Assignee: |
Longhorn Vaccines and Diagnostics,
LLC
Bethesda
MD
|
Family ID: |
1000004827959 |
Appl. No.: |
16/812322 |
Filed: |
March 8, 2020 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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15983215 |
May 18, 2018 |
10596250 |
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16812322 |
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15447972 |
Mar 2, 2017 |
10004799 |
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15983215 |
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15205476 |
Jul 8, 2016 |
9777045 |
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15447972 |
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14473605 |
Aug 29, 2014 |
9388220 |
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15205476 |
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12199729 |
Aug 27, 2008 |
8821885 |
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14473605 |
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13750771 |
Jan 25, 2013 |
9598462 |
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15447972 |
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60968145 |
Aug 27, 2007 |
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61591113 |
Jan 26, 2012 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 2039/55566
20130101; A61K 2039/5254 20130101; C12N 2760/16134 20130101; A61K
2039/525 20130101; C12N 2760/16161 20130101; C12N 2760/16122
20130101; C07K 16/1018 20130101; A61K 39/145 20130101; C07K 16/1289
20130101; C07K 14/35 20130101; C07K 14/005 20130101; A61K 2039/6043
20130101; A61K 2039/53 20130101; A61K 2039/70 20130101; C07K
2319/40 20130101; C12N 7/00 20130101; A61K 2039/6068 20130101; A61K
2039/6037 20130101; C07K 2319/42 20130101; C07K 14/33 20130101;
A61K 39/12 20130101; A61K 2039/5252 20130101; A61K 2039/545
20130101; A61K 39/04 20130101; A61K 2039/575 20130101; A61K
2039/543 20130101 |
International
Class: |
A61K 39/145 20060101
A61K039/145; C07K 14/35 20060101 C07K014/35; C07K 14/005 20060101
C07K014/005; C07K 16/12 20060101 C07K016/12; C07K 16/10 20060101
C07K016/10; A61K 39/04 20060101 A61K039/04; C12N 7/00 20060101
C12N007/00; A61K 39/12 20060101 A61K039/12; C07K 14/33 20060101
C07K014/33 |
Claims
1. A method of treating or preventing an infection of an influenza
virus comprising: providing an immunogenic composition that
contains a peptide which comprises an amino acid sequence that is
at least 95% identical to any one of SEQ ID NOs: 1-116; and a
T-cell stimulating epitope; and administering the immunogenic
composition to a patient to generate an immunological response to
the infection.
2. The method of claim 1, wherein the influenza virus comprises one
or more of H1N1, H1N7, H2N2, H3N1, H3N2, H3N8, H5N1, H5N2, H5N3,
H5N8, H5N9, H7N1, H7N2, H7N3, H7N4, H7N7 or H9N2.
3. The method of claim 1, wherein the peptide comprises the
sequence of one or more of SEQ ID NO 1, SEQ ID NO 2, SEQ ID NO 6,
SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, SEQ ID NO 12, SEQ ID NO 13,
SEQ ID NO 14, SEQ ID NO 15, SEQ ID NO 16, SEQ ID NO 17, SEQ ID NO
18, SEQ ID NO 19, SEQ ID NO 20, SEQ ID NO 47, SEQ ID NO 52, SEQ ID
NO 53, and SEQ ID NO 54.
4. An immunogenic composition that contains a peptide which
comprises an amino acid sequence that is at least 95% identical to
any one or more of SEQ ID NOs: 1-116; and a T-cell stimulating
epitope.
5. The composition of claim 4, which further contains an
adjuvant.
6.-45. (canceled)
Description
REFERENCE TO RELATED APPLICATIONS
[0001] This application is a Continuation of U.S. application Ser.
No. 15/983,215 filed May 18, 2018, which issued as U.S. Pat. No.
10,596,250 on Mar. 24, 2020, which is a Continuation of U.S.
application Ser. No. 15/447,972 filed Mar. 2, 2017, which issued as
U.S. Pat. No. 10,004,799 on Jun. 26, 2018, which is a
Continuation-in-Part of U.S. application Ser. No. 15/205,476 filed
Jul. 8, 2016, which issued as U.S. Pat. No. 9,777,045 on Oct. 3,
2017, which is a Continuation of U.S. application Ser. No.
14/473,605 filed Aug. 29, 2014, which issued as U.S. Pat. No.
9,388,220 Jul. 12, 2016, which is a Continuation of U.S.
application Ser. No. 12/199,729 filed Aug. 27, 2008, which issued
as U.S. Pat. No. 8,821,885 Sep. 2, 2014, and claims priority to
U.S. Provisional Application No. 60/968,145 filed Aug. 27, 2007,
and a Continuation of U.S. application Ser. No. 13/750,771 filed
Jan. 25, 2013 which issued as U.S. Pat. No. 9,598,462 Mar. 21,
2017, and claims priority to U.S. Provisional Application No.
61/591,113 filed Jan. 26, 2012, each of which is entirely
incorporated by reference.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which
has been submitted electronically in ASCII format and is hereby
incorporated by reference in its entirety. Said ASCII copy, created
on Sep. 30, 2019, is named 3022.015USCP03_SL.txt and is 39 kbytes
in size.
BACKGROUND
1. Field of the Invention
[0003] The present invention is directed to composite antigens
composed of a plurality of epitopes, and to tools and methods for
generating an immune response with the composite antigens of the
invention. The invention is also directed to compositions
comprising composite antigenic sequences derived from multiple
pathogens for the development of novel vaccines and to the vaccines
developed.
2. Description of the Background
[0004] Microbial and viral pathogens are a primary source of
infectious disease in animals. Pathogens and their hosts constantly
adapt to one another in an endless competition for survival and
propagation. Certain pathogens have become enormously successful at
infecting mammalian hosts and surviving exposure to the host immune
response, even over periods of years or decades. One example of an
extremely successful mammalian pathogen is the influenza virus.
[0005] Influenza viruses are etiologic agents for a contagious
respiratory illness (commonly referred to as the flu) that
primarily affects humans and other vertebrates. Influenza is highly
infectious and an acute respiratory disease that has plagued the
human race since ancient times. Infection is characterized by
recurrent annual epidemics and periodic major worldwide pandemics.
Influenza virus infection can cause mild to severe illness, and can
even lead to death. Every year in the United States, 5 to 20
percent of the population, on average, contracts the flu with more
than 200,000 hospitalizations from complications and over 36,000
deaths. Because of the high disease-related morbidity and
mortality, direct and indirect social economic impacts of influenza
are enormous. Four pandemics occurred in the last century, together
causing tens of millions of deaths worldwide.
[0006] Influenza virus spreads from host to host through coughing
or sneezing. Airborne droplets are the primary transmission vectors
between individuals. In humans, the virus typically spreads
directly from person to person, although persons can also be
infected from indirect contact with surfaces harboring the virus.
Infected adults become infectious to others beginning as little as
one day before primary symptoms of the disease develop. Thereafter,
these persons remain infectious for up to 5 days or more after.
Uncomplicated influenza illness is often characterized by an abrupt
onset of constitutional and respiratory symptoms, including fever,
myalgia, headache, malaise, nonproductive cough, sore throat,
rhinitis, or a combination of one or more of these symptoms.
[0007] Currently, the spread of pathogenic influenza virus is
controlled in animal populations by vaccination and/or treatment
with one or more anti-viral compounds. Vaccines containing
inactivated influenza virus or simply influenza antigen are
currently in use worldwide and especially promoted for use by
high-risk groups such as infants, the elderly, those without
adequate health care and immunocompromised individuals. Most all
viruses for vaccine use are propagated in fertile hen's eggs,
inactivated by chemical means, and the antigens purified. The
vaccines are usually trivalent, containing representative influenza
A viruses (H1N1 and H3N2) and influenza B strains. The World Health
Organization (WHO) regularly updates the specific strains targeted
for vaccine development to those believed to be most prevalent and
thereby maximize overall world efficacy. During inter-pandemic
periods, it typically takes eight months or more before an updated
influenza vaccine is ready for market. Historically, viral
pandemics are spread to most continents within four to six months,
and future viral pandemics are likely to spread even faster due to
increased international travel. It is likely inevitable that an
effective vaccine made by conventional means will be unavailable or
in very short supply during the first wave of any future widespread
outbreak or pandemic.
[0008] Pandemic flu can and does arise at any time. Although the
severity of the next Influenza pandemic cannot be accurately
predicted, modeling studies suggest that the impact of a pandemic
on the United States, and the world as a whole, would be
substantial. In the absence of any control measures (vaccination or
drugs), it has been estimated that in the United States a
"medium-level" pandemic could cause: 89,000 to 207,000 deaths;
314,000 and 734,000 hospitalizations; 18 to 42 million outpatient
visits; and another 20 to 47 million people being sick. According
to the Centers for Disease Control and Prevention (CDC) (Atlanta,
Ga., USA), between 15 percent and 35 percent of the U.S. population
could be affected by an influenza pandemic, and the economic impact
could range between approximately $71 and $167 billion.
[0009] Vaccines capable of producing a protective immune response
have been produced in the last half century. These include whole
virus vaccines, split-virus vaccines, and surface-antigen vaccines
and live attenuated virus vaccines. While formulations of any of
these vaccine types are capable of producing a systemic immune
response, live attenuated virus vaccines have the advantage of also
being able to stimulate local mucosal immunity in the respiratory
tract.
[0010] With the continual emergence (or re-emergence) of different
influenza strains, new influenza vaccines are continually being
developed. Because of the rapid mutation rate among Influenza
viruses, it has been extremely difficult and at times not possible
to identify the antigenic moieties of the emergent virus strains in
sufficient time to develop a suitable vaccine. Thus, polypeptides
and polynucleotides of newly emergent or re-emergent virus strains
(especially sequences of antigenic genes) are highly desirable.
[0011] Influenza is typically caused by infection of two genera of
influenza viruses: Influenzavirus A and Influenzavirus B. The third
genus of influenza viruses, Influenzavirus C, exists as a single
species, influenza C virus, which causes only minor common
cold-like symptoms in susceptible mammals. Infections by influenza
A virus and influenza B virus are typically initiated at the
mucosal surface of the upper respiratory tract of susceptible
mammals. Viral replication is primarily limited to the upper
respiratory tract but can extend to the lower respiratory tract and
cause bronchopneumonia that can be fatal.
[0012] Influenza A virus, in particular, has many different
serotypes. Presently, there are sixteen known variations of HA (the
hemaglutination antigen which is involved in virus to cell binding)
and nine known variations of NA (the neuraminidase antigen which is
involved in viral release) within influenza A viruses, thus
yielding 144 possible "HN" serotypes of influenza A virus based on
variations within these two proteins alone. Only a small number of
these combinations are believed to be circulating within
susceptible populations at any given time. Once a new influenza
strain or serotype emerges and spreads, the historical pattern is
that it becomes established within the susceptible population and
then moves around or "circulates" for many years causing seasonal
epidemics of the Flu.
[0013] Three genera of influenza viruses currently comprise the
Orthomyxoviridae Family: Influenza virus A, Influenza virus B, and
Influenza virus C. Each of these genera contains a single species
of influenza virus: The genus Influenza virus A consists of a
single species, influenza A virus, which includes all of the
influenza virus strains currently circulating among humans,
including, for example, but not limited to, H1N1, H1N2, H2N2, H3N1,
H3N2, H3N8, H5N1, H5N2, H5N3, H5N8, H5N9, H7N1, H7N2, H7N3, H7N4,
H7N7, H9N2, and H10N7 serotypes. Exemplary influenza A viral
strains include, but are not limited to, A/Aichi/2/68,
A/Alaska/6/77, A/Alice, A/Ann Arbor/6/60, A/Bayern/7/95,
A/Beijing/352/89, A/Beijing/353/89, A/Bethesda/1/85,
A/California/10/78, A/Chick/Germany/N/49, A/Chile/1/83,
A/Denver/1/57, A/Dunedin/6/83, A/Equine/Miami/1/63, A/FM/1/47,
A/Great Lakes/0389/65, A/Guizhou/54/89, A/Hong Kong/77, A/Hong
Kong/8/68, A/Hong Kong/483/97, A/Johannesburg/33/94,
A/Kawasaki/9/86, A/Kiev/59/79, A/Korea/1/82, A/Korea/426/68,
A/Leningrad/13/57, A/Los Angeles/2/87, A/MaI/302/54,
A/Memphis/8/88, A/Nanchang/933/95, A/New Jersey/8/76, A/NT/60/68,
A/NWS/33, A/Peking/2/79, A/Port Chalmers/1/73, A/PR/8/34,
A/Shanghai/11/87, A/Shanghai/16/89, A/Shanghai/31/80,
A/Singapore/1/57, A/Singapore/6/86, A/South Carolina/1/181918,
A/Swine/1976/31, A/Swine/Iowa/15/30, A/Swine/New Jersey/8/76,
A/Sydney/5/97, A/Taiwan/1/86, A/Taiwan/1/86A1, A/Texas/35/91,
A/Texas/36/91, A/USSR/90/77, A/Victoria/3/75, A/Vietnam/1203/04,
A/Washington D.C./897/80, A/Weiss/43, A/WS/33, A/WSN/33,
A/Wuhan/359/95, A/Wyoming/1/87, and A/Yamagata/32/89, as well as
derivatives, variants, and homologs thereof.
[0014] The genus Influenza virus B consists of a single species,
influenza B virus, of which there is currently only one known
serotype. Influenza B virus is almost exclusively a human pathogen,
but is significantly less common and less genetically diverse than
influenza A strains. Because of this limited genetic diversity,
most humans acquire a certain degree of immunity to influenza B
virus at an early age; however, the mutation frequency of the virus
is sufficiently high enough to prevent lasting immunity by most
humans, but not high enough to permit pandemic infection by
influenza B virus across human populations. Exemplary influenza B
viral serotypes include, but are not limited to, B/Allen/45, B/Ann
Arbor/1/86, B/Bangkok/163/90, B/Beijing/184/93, B/Brigit,
B/GL/1739/54, B/Hong Kong/330/2001, B/Hong Kong/5/72, B/Lee/40,
B/Maryland/1/59, B/Mass/3/66, B/Oman/16296/2001, B/Panama/45/90,
B/R22 Barbara, B/R5, B/R75, B/Russia/69, B/Shandong/7/97,
B/Sichuan/379/99, B/Taiwan/2/62, B/Tecumseh/63/80, B/Texas/1/84,
B/Victoria/2/87, and B/Yamagata/16/88, as well as derivatives,
variants, and homologs thereof.
[0015] The genus Influenza virus C also consists of a single
species, denoted influenza C virus, of which there is also
currently only one known serotype. This serotype is known to infect
both primates and porcines, and while infections of influenza C
virus are rare, the resulting illness can be severe. Epidemics of
influenza C virus are not uncommon in exposed populations, however,
due to its rapid transmissibility in humans having close
contact.
[0016] Polynucleotide and polypeptide sequences from each of these
strains are contained within the publicly-available databases of
the National Center for Biotechnology Information (National Library
of Medicine, National Institutes of Health, Bethesda, Md., USA),
and viral stocks may be obtained from the American Type Culture
Collection (Manassas, Va., USA), or are otherwise publicly
available.
[0017] Human influenza virus usually refers to influenza virus
serotypes that are transmissible among humans. There are only three
known influenza A virus HN serotypes that have circulated widely
among humans in recent times: H1N1, H2N2, and H3N2. Many humans
have acquired at least some level of immunity to these subtypes.
All Influenza viruses, however, are known to mutate and change
frequently. Influenza viruses are known to infect waterfowl and
swine and to circulate among those hosts forming a breeding ground
for new subtypes and strains separate from human populations.
Because many serotypes (and particularly newly-arising subtypes)
have a zero or low prevalence in human populations, there is little
or no natural immunity against them in human populations. Such a
population is referred to as being "naive" to such serotypes.
Accordingly, Influenza viruses might be expected to adapt over time
to generate one or more highly virulent strains that will infect
and spread catastrophically among naive humans, as has been widely
reported in the mainstream press.
[0018] The highly-virulent influenza H5N1 subtype (publicly
referred to as the bird flu virus), for example, has been reported
as having mutated sufficiently to become transmissible from avian
hosts to humans. As this subtype has been limited to infecting
avian populations in the past, there is little or no legacy of
infection to have generated immunity within the human population.
Thus, the human population is expected to be highly susceptible to
H5N1.
[0019] To date, the H5N1 serotype does not appear to have mutated
sufficiently to become efficiently transmitted from human to human.
Nonetheless, because influenza viruses are constantly adapting,
there is concern that H5N1 virus or another virulent influenza
strain or serotype will arise that will be able to infect humans
and spread easily from one person to another. It has been commonly
suggested that if H5N1 virus were to gain the capacity to spread
easily from person to person, a worldwide outbreak of disease
(i.e., pandemic) would likely begin, resulting in millions of
deaths.
[0020] Annual influenza outbreaks occur as a result of "antigenic
drift." Antigenic drift is caused by mutations within antigenic
(i.e., immunity stimulating) portions of viral proteins within
viral subtypes circulating in host populations that alter the
host's ability to recognize and defend effectively against the
infecting virus, even when the virus has been circulating in the
community for several years. The antigenic drift that diminishes
existing immunity in a host population generally occurs within
so-called immunodominant antigens or regions.
[0021] Immunodominant antigens are those antigens belonging to a
pathogen that are the most-easily and most-quickly recognized by
the host immune system and, consequently, account for the vast
majority of immune response to the invading pathogen. Typically,
immunodominant antigens exist within regions of the pathogen that
are most exposed to the environment, i.e., are on the external
surfaces or on protruding elements of the pathogen, and so are most
readily accessible to the host immune system.
[0022] In the case of influenza, the immunodominant HA and NA
proteins protrude from the central capsid of the viral particle,
and so they tend to interact most strongly with the host's internal
environment and dominate the host immune response. Mutations
occurring in the microbial genome that protect the microbe from the
host immune system, these mutations are most readily found to
affect the immunodominant antigens.
[0023] Conversely, non-immunodominant antigens are those that are
capable of raising a host immune response but account for only a
small amount of the total immune response. This is thought to
happen because the non-immunodominant antigens are at least
partially shielded from the host immune system, as in the case of
an antigen that is located in a cleft or fold of the microbial
surface or is surrounded by protruding elements of the microbe. In
the case of influenza, non-immunodominant antigens occurring near
the capsid surface are shielded from the host immune system by the
immunodominant HA and NA spikes protruding from the surface.
Non-immunodominant antigens tend to show less mutation in response
to host immune pressure than do immunodominant antigens.
[0024] Antigenic shift occurs when there is an abrupt or sudden,
major change in a virus. Antigenic shift is typically caused by the
occurrence of new combinations of the HA and/or NA proteins on the
surface of the virus, i.e., the creation of a new Influenza
subtype. The appearance of a new influenza A virus subtype, to
which most of the world's population is naive, is the first step
toward a pandemic. If the new Influenza subtype also has the
capacity to spread easily from person to person, then a full-blown
pandemic may be expected resulting in a global influenza outbreak
infecting millions of humans.
[0025] The CDC and the leading authorities on disease prevention in
the world recommend the single best way of preventing the flu is
through annual vaccination. Conventional vaccines however,
typically target the HA and NA antigens, and have been neither
universally protective nor 100 percent effective at preventing the
disease. Antigenic shift prevents flu vaccines from being
universally protective or from maintaining effectiveness over many
years. The ineffectiveness of conventional vaccines may also be
due, in part, to antigenic drift and the resulting variation within
antigenic portions of the HA and NA proteins most commonly
recognized by the immune system (i.e., immunodominant antigens). As
a result, many humans may find themselves susceptible to the flu
virus without an effective method of treatment available since
influenza is constantly improving its resistant to current
treatments. This scenario is particularly concerning with respect
to the H5N1 virus, which is highly virulent but for which there is
currently no widely available commercial vaccine to immunize
susceptible human populations.
[0026] Currently, flu vaccines are reformulated each year due to
the yearly emergence of new strains, and generally induce limited
immunity. In addition, to achieve a protective immune response,
some vaccines are administered with high doses of antigen. This is
particularly true for H5N1 vaccines. In addition, influenza
vaccines, including H5N1 vaccines, typically present epitopes in
the same order as the epitopes are found in nature, generally
presenting as whole-viral proteins; consequently, relatively large
amounts of protein are required to make an effective vaccine. As a
result, each administration includes an increased cost associated
with the dose amount, and there is increased difficulty in
manufacturing enough doses to vaccinate the general public.
Further, the use of larger proteins elevates the risk of
undesirable immune responses in the recipient host.
[0027] Accordingly, it would be advantageous to administer a
vaccine that provides protection against an infection over a broad
range of different strains and/or variations of a pathogen, and a
vaccine that is effective against multiple pathogens. It would also
be advantageous to administer a single or limited number of
vaccinations that would provide effective protection across a
selection of different pathogens and a vaccine that could be
effective in those individuals with limited immune system function.
Such vaccines would be useful to treat many individuals and
populations and may be useful to compliment conventional vaccines,
all to provide comprehensive protection to as many individuals as
possible against existing as well as new and emerging pathogens
across a population.
SUMMARY OF THE INVENTION
[0028] The present invention provides new and useful compositions,
as well as tools and methods for generating an immune response. In
particular, the invention provides vaccines and methods developed
from multiple antigenic regions of one or more pathogens.
[0029] One embodiment of the invention is directed to a composite
antigen comprising a peptide with contiguous amino acid sequence
derived from a plurality of antigenic epitopes of one or more
pathogens that induces an immune response in a mammal that is
protective against infection by the one or more pathogens.
Preferably the plurality of epitopes contains one or more composite
epitopes. Preferably the composite antigen contains a plurality of
antigenic epitopes, comprising one or more repetitions of a same
epitope, one or more repetitions of different epitopes, one or more
repetitions of composite epitopes, or a combination thereof. Also
preferably, the amino acid sequence of at least one epitope of the
composite antigen does not exist naturally. Composite antigens can
be used to treat or preferably prevent infection and disease
associated with one or more pathogens including but not limited to
viruses, bacteria, parasites, yeast, fungi, or a combination
thereof. Preferably the pathogen is an influenza virus and the one
or more antigenic epitopes are amino acid sequences of M1, M2, HA,
NA, PB1, or PB2 protein, or a combination thereof. Exemplary
composite sequences include, but are not limited to, SEQ ID NOs 4,
5, 8, 19, 20, 52, 53, 56 and 54, and SEQ ID NO 16, 65, 66, 67, 70
and 73.
[0030] Another embodiment of the invention is directed to composite
antigens comprising an amino acid sequence containing amino acids
that are in common between sequences of epitopes of multiple
conserved regions, and the amino acids that differ between the
sequences of epitopes of multiple conserved regions. Preferably,
the amino acid sequence has the formula An1BCAn2, wherein A
represents the amino acids that are in common between sequences of
epitopes of multiple conserved regions, B and C represent the amino
acids that differ between the sequences of epitopes of multiple
conserved regions, wherein A, B, and C are naturally occurring
amino acids, N1 and N2 total to less than 25, and the number of B
and C amino acids is less than 3. Exemplary composite sequences
include, but are not limited to SEQ ID NO. 6, 7, 21, 22, 54, 55, 58
or 59. Preferably the composite antigen contains multiple conserved
regions of a peptide sequence derived from multiple serotypes of a
same pathogen. Preferably the pathogen is influenza virus.
[0031] Another embodiment of the invention is directed to an
antibody that is specifically reactive to the composite antigen of
the invention.
[0032] Another embodiment of the invention is directed to
polynucleotides that encode composite antigens of the
invention.
[0033] Another embodiment of the invention is directed to methods
for generating an immune response in a mammal comprising
administering to the mammal the composite antigen of the invention.
Preferably the immune response generated is protective against a
number of different strains, serotypes or species of the one or
more pathogens.
[0034] Another embodiment of the invention is directed to a vaccine
comprising the composite antigen of the invention. Preferably the
composite antigen is has the formula An1BCAn2, wherein A represents
the amino acids that are in common between sequences of epitopes of
multiple conserved regions, B and C represent the amino acids that
differ between the sequences of epitopes of multiple conserved
regions, wherein A, B, and C are naturally occurring amino acids,
N1 and N2 total to less than 25, and the number of B and C amino
acids is less than 3.
[0035] Other embodiments and advantages of the invention are set
forth in part in the description, which follows, and in part, may
be obvious from this description, or may be learned from the
practice of the invention.
DESCRIPTION OF THE DRAWINGS
[0036] FIG. 1 Summary of ELISA antisera titers of peptides (or Pep)
3, 6, 9, 10 and 11 of H3N2 Influenza virus (Wuhan).
[0037] FIG. 2 Antisera mean ODs of mice immunized with different
peptides of H3N2 Influenza virus (Wuhan 1:40).
[0038] FIG. 3 Mean ODs of antisera (1:100) on virus and peptide
following immunization with Pep 6
[0039] FIG. 4 Antisera titers for mice immunized with Pep 9 on H1N1
(Caledonia) virus at various dilutions.
[0040] FIG. 5A Assay of Pep 9 sera on fresh Wuhan coating.
[0041] FIG. 5B Assay of Pep 9 sera on fixed Wuhan.
[0042] FIG. 5C Assay of Pep 9 sera on fresh Caledonia coating.
[0043] FIG. 5D Assay of Pep 9 sera on fixed Caledonia coating.
[0044] FIG. 6 Antisera titers from mice immunized with Pep 9
captured on fixed vs. Fresh coatings of Wuhan and Caledonia.
[0045] FIG. 7 Antisera titers for mice immunized with Pep 11 on
H5N1 at various dilutions.
[0046] FIG. 8-1 Additional sequences and related information.
[0047] FIG. 8-2 Additional sequences and related information.
[0048] FIG. 8-3 Additional sequences and related information.
[0049] FIG. 8-4 Additional sequences and related information.
[0050] FIG. 8-5 Additional sequences and related information.
[0051] FIG. 8-6 Additional sequences and related information.
DESCRIPTION OF THE INVENTION
[0052] Vaccinations and vaccines are often the best mechanism for
avoiding an infection and preventing the spread of debilitating and
dangerous pathogens. With respect to viral infections and many
bacterial infections, vaccinations are often the only effective
option as treatment options are few and those that are available
provide only limited effectiveness. Conventional vaccinations
require a priori understanding or general identification of the
existing antigenic regions of the pathogen. The pathogen itself is
propagated and a suitable vaccine developed from heat-killed or
otherwise attenuated microorganisms. Alternatively, an antigen or
collection of antigens is identified that will generate a
protective immune response upon administration. The need for a
vaccine is especially urgent with respect to preventing infection
by certain bacteria and viruses. Many bacteria and especially
certain viruses mutate constantly often rendering the vaccine
developed to the prior or originating bacteria or virus useless
against the new strains that emerge. As a consequence, vaccines
against infections are reformulated yearly and often administered
at fairly high doses. The manufacturing costs are high and
administering vaccines against pose a great many complications and
associated risks to patients.
[0053] It has been surprisingly discovered that an effective
vaccine can be produced from a composite antigen of the invention.
Composite antigens are antigens that contains or are derived from a
plurality of antigenic regions of a pathogen. Composite antigens of
the invention may contain one epitope that represents a combination
of conserved regions of two or more epitopes (e.g., the composite
epitope as outlined herein), or a plurality of immunologically
responsive regions derived from one or multiple sources (e.g.,
virus particles, parasites, bacteria, fungi, cells). These
immunological regions are amino acid sequences or epitopes that are
representative of sequences found at those antigenic regions of a
pathogen or other antigen associated with an infection or a disease
or, importantly, associated with stimulation of the immune system
to provide protection against the pathogen.
[0054] One embodiment of the invention is directed to composite
antigens. Composite antigens of the invention contain non-naturally
occurring amino acid sequences that do not exist in nature and are
otherwise artificially constructed. Each sequence of a composite
antigen contains a plurality of immunologically responsive regions
or epitopes of a pathogen, which are artificially arranged,
preferably along a single amino acid sequence. The plurality may
contain multiples of the same epitope, although not in a naturally
occurring order, or multiples of a variety of different epitopes
from one or more pathogens. Epitopes may be identical to known
immunological regions of a pathogen, or entirely new constructs
that have not previously existed and therefore artificially
constructed. Preferably, the composite antigen of the invention
induces killer T-cell (Tc or CTL) responses, helper T-cell (Tx)
responses, and specific antibody production in an inoculated
mammal.
[0055] A "composite" antigen is an engineered, artificially created
antigen made from two or more constituent epitopes, such that the
resulting composite antigen has physical and/or chemical properties
that differ from or are additive of the individual constituent
epitopes. Preferable the composite antigen, when exposed to the
immune system of a mammal, is capable of simultaneously generating
an immunological response to each of the constituent epitope of the
composite and preferably to a greater degree (e.g., as measurable
from a cellular or humoral response to an identified pathogen) than
the individual constituent epitopes. In addition, the composite
antigen provides the added function of generating a protective
immunological response in a patient when used as a vaccine and
against each of the constituent epitopes. Preferably, the composite
has the additional function of providing protection against not
only the pathogens from which the constituents were derived, but
related pathogens as well. These related pathogenic organisms may
be strains or serotypes of the same species of organism, or
different species of the same genus of organism, or different
organisms entirely that are only related by a common epitope.
[0056] Another embodiment of the invention is directed to isolated
epitopes. Isolated epitopes are regions obtained or derived from a
protein or peptide of a pathogen that elicit a robust immunological
response when administered to a mammal. Preferably, that robust
response provides the mammal with an immunological protection
against developing disease from exposure to the pathogen. A
preferred example of an isolated epitope is a composite epitope,
which is one artificially created from a combination of two or more
highly conserved, although not identical, amino acid sequences of
two or more different, but otherwise related pathogens. The
pathogens may be of the same type, but of a different strain,
serotype, or species or other relation. The composite epitope
contains the conserved region that is in common between the related
epitopes, and also contains the variable regions which differ. The
sequences of a composite epitope that represents a combination of
two conserved, but not identical sequences, may be illustrated as
follows:
TABLE-US-00001 Sequence of Epitope 1 . . . AAAAABAAAAA . . .
Sequence of Epitope 2 . . . AAAAACAAAAA . . . Composite Epitope . .
. AAAAABCAAAAA . . .
wherein, "A" represents the amino acids in common between the two
highly conserved epitopes, "B" and "C" represent the amino acids
that differ, respectively, between two epitopes, each of "A"," "B"
and "C" can be any amino acid and any number of amino acids.
Preferably the conserved region contains about 20 or less amino
acids on each side of the variable amino acids, preferably about 15
or less, preferably about 10 or less, preferably about 8 or less,
preferably about 6 or less, and more preferably about 4 or less.
Preferably the amino acids that vary between two similar but not
identical conserved regions are 5 or less, preferably 4 or less,
preferably 3 or less, preferably 2 or less, and more preferably
only 1.
[0057] A "composite epitope," similar to the composite antigen, is
an engineered, artificially created single epitope made from two or
more constituent epitopes, such that the resulting composite
epitope has physical and/or chemical properties that differ from or
are additive of the constituent epitopes. Preferable the composite
epitope, when exposed to the immune system of a mammal, is capable
of simultaneously generating an immunological response to each of
the constituent epitopes of the composite and preferably to a
greater degree than that achieved by either of the constituent
epitopes individually. In addition, the composite epitope provides
the added function of generating a protective immunological
response in a patient when used as a vaccine and against each of
the constituent epitopes. Preferably, the composite has the
additional function of providing protection against not only the
pathogens from which the constituents were derived, but related
pathogens as well. These related pathogenic organisms may be
strains or serotypes of the same species of organism, or different
species of the same genus of organism, or different organisms
entirely that are only related by a common epitope.
[0058] Composite epitopes of the invention are entirely artificial
molecules that do not otherwise exist in nature and to which an
immune system has not been otherwise exposed. Preferably, these
conserved immunological regions that are combined as a composite
epitope represent immunologically responsive regions of proteins
and/or polypeptides that are highly conserved between related
pathogens. Although a vaccine can be developed from a single
composite epitope, in many instances the most effective vaccine may
be developed from multiple, different composite epitopes.
[0059] Accordingly, composite antigens of the invention may contain
one or more composite epitopes and/or one or more known epitopes to
provide an effective vaccine. Although composite antigens may
comprise a single composite epitope, a composite antigen would not
comprise only a single known epitope. Preferably, the immunological
response achieved from a vaccination with a composite antigen, or
group of composite antigens, provides protection against infection
caused by the original strains from which the sequence of the
composite antigen was derived, and also provides immunological
protection against other strains, serotypes and/or species that
share one or more of the general conserved regions represented in
the composite antigen. Thus, the resulting immune response achieved
from a vaccination with a composite antigen is more broadly
protective than can be achieved from a conventional single antigen
vaccination against multiple strains, serotypes, and species or
otherwise related pathogens regardless of antigenic drift that may
take place in the evolution of the pathogen. Preferably, vaccines
developed from composite antigens of the invention avoid any need
for repeated or annual vaccinations, the associated complications
and expenses of manufacture, and the elevated risks to the patient.
These vaccines are useful to treat individuals and populations,
thereby preventing infection, mortality and pandemics, and are
useful to compliment conventional vaccines.
[0060] As discussed herein, the composite antigen preferably
comprises a single chain of amino acids with a sequences derived
from one or more composite eptiopes, one or more composite epitopes
plus one or more known epitopes, or a plurality of known epitopes,
that may be the same or different. Epitope sequences may be
repeated consecutively and uninterrupted along a composite sequence
or interspersed among other sequences that may be single or a few
amino acids as spacers or sequences that encode peptides
(collectively spacers), and may be nonimmunogenic or immunogenic
and capable of inducing a cellular (T cell) or humoral (B cell)
immune response in a mammal. Peptides sequence from unrelated
microbes may be combined into a single composite antigen. For
example, viral sequences of selected immunoresponsive peptides may
be interspersed with conserved sequences or epitopes selected from
other microbes, such as, for example, bacteria such as S.
pneumococcus, P. auriginosa or S. aureus. Preferred viral proteins,
from which preferred epitopes may be selected, include, but are not
limited to the influenza virus proteins PspA, PspC, HA, NA, M2e, H.
influenza protein D, and coagulase.
[0061] An epitope of the composite antigen may be of any sequence
and size, but is preferable composed of natural amino acids and is
more than 5 but less than 35 amino acids in length, preferably less
than 30, preferably between 5 and 25 amino acids in length,
preferably between 8 and 20 amino acids in length, and more
preferably between 5 and 15 amino acids in length. Composite
antigens preferably contain any number of composite and/or other
epitopes. The most effective number of epitopes of a composite
antigen against a particular pathogen, pathogen family, or group of
pathogens may be determined by one skilled in the art from the
disclosures of this application and using routine testing
procedures. Composite antigens may be effective with one epitope,
preferably with 2 or more, 3 or more 4 or more, 5 or more or
greater. Optionally, composite antigens may include one or more
spacers between epitopes which may be sequences of antigenic
regions derived from the same or from one or more different
pathogens, or sequences that serve as immunological primers or that
otherwise provide a boost to the immune system. That boost may be
generated from a sequence of amino acids that are known to
stimulate the immune system, either directly or as an adjuvant. In
one preferred embodiment, composite antigens useful to generate an
immunological response against influenza virus comprise epitopes of
HA and/or NA proteins, and/or new epitopes derived from similar
conserved regions of different serotypes of influenza virus. Also
preferred are composite antigens containing epitopes of proteins of
Mycobacterium tuberculosis and Clostridium tetani, and/or new
epitopes derived from similar conserved regions of different
serotypes of these bacteria.
[0062] Another embodiment of the invention is directed to a
contiguous sequence of one or more epitopes, which may comprise
composite and/or known epitopes, from one or more pathogens in a
sequence that does not exist naturally and must be artificially
constructed. Preferably, a contiguous sequence of the invention
contains one or more composite epitopes, which is a combination of
the sequences of the conserved regions of epitopes that are common
to multiple pathogens plus those amino acids that differ between
the two conserved regions. For example, where two pathogens contain
similar conserved regions that differ by only a single amino acid,
the composite sequences would include the conserved region amino
acids and each of the amino acids that differ between the two
regions as discussed herein.
[0063] It is also preferable that a composite antigen of the
invention contain a plurality of repeated epitopes and, optionally,
epitopes conjugated with linker regions between or surrounding each
epitope, and the plurality of epitopes be the same or different.
Preferred linkers include amino acid sequences of antigenic regions
of the same or of different pathogens, or amino acids sequences
that aid in the generation of an immune response. Preferred
examples include, but are not limited to any of the various
antigenic regions of bacteria such as, but not limited to
tuberculosis and virus such as, but not limited to influenza. It is
also preferred that composite antigens contain epitopes that
generate a T cell response, a B cell response, or both in
conjunction with a specific response to the pathogen.
[0064] Another embodiment of the invention is directed to
immunizing animals with the composite antigens of the invention.
Antisera obtained from the immunized animals are reactive against
the pathogens from which the composite antigen was derived. Another
embodiment of the invention is therefore antisera obtained from the
immunized animals, which may be further purified for testing or
utilized therapeutically for administration to another mammal and
thereby provides protection against infection from the one or more
pathogens. It is also preferred that the antisera obtained provide
a broad protection, not just against the pathogens from which the
composite antigen was derived, but also from additional pathogens
that may differ by strain, serotype, or even species.
[0065] Another embodiment of the invention is a vaccine composed of
the composite antigen or antisera developed from the composite
antigen of the invention. Preferably, the vaccines of the invention
are less susceptible to variation of antigenicity due to antigenic
shift of pathogens which reduces or eliminates the need for annual
or repeated vaccination to maintain protection of patient
populations against potential outbreaks of infection from new viral
isolates. In addition, the vaccines of the invention generally and
advantageously provide increased safety considerations, both in
their manufacture and administration (due in part to a
substantially decreased need for repeated administration), a
relatively long shelf life in part due to minimized need to
reformulate due to strain-specific shift and drift, an ability to
target immune responses with high specificity for particular
microbial epitopes, and an ability to prepare a single vaccine that
is effective against multiple pathogens, each of which may be a
different but know strain or species of the same pathogen. The
invention encompasses antigenic and antibody compositions, methods
of making such compositions, and methods for their use in the
prevention, treatment, management, and/or prophylaxis of an
infection. The compositions disclosed herein, as well as methods
employing them, find particular use in the treatment or prevention
of viral, bacterial, parasitic and/or fungal pathogenesis and
infection using immunogenic compositions and methods superior to
conventional treatments presently available in the art.
[0066] Another embodiment of the invention is directed to methods
of preventing or controlling infection, such as, for example, an
outbreak of viral, parasitic, fungal or bacterial infection,
preferably but not limited to an influenza virus and/or a
tuberculosis bacterial infection, in a selected mammalian
population. The method includes at least the step of providing an
immunologically effective amount of one or more of the disclosed
immunogenic or vaccine compositions to a susceptible or an at-risk
member of the population, for a time sufficient to prevent, reduce,
lessen, alleviate, control, or delay the outbreak of such an
infection in the general population.
[0067] Another embodiment of the invention is directed to methods
for producing a protective immune response against infection, for
example by influenza virus, in a mammal in need thereof. Such a
method generally includes a step of providing to a mammal in need
thereof, an immunologically-effective amount of one or more of the
immunogenic compositions disclosed herein under conditions and for
a time sufficient to produce such a protective immune response
against one or more species, strains, or serotypes of an infectious
organism. Additionally, the invention also provides a method for
administering a prophylactic antiviral or antimicrobial composition
to at least a first cell, tissue, organ, or organ system in a
mammal that generally involves providing to such a mammal a
prophylactically-effective amount of at least a first immunogenic
composition as disclosed herein.
[0068] Another embodiment of the invention is directed to an
immunogenic composition comprising the composite antigens of the
invention having one or more repeated peptide sequences, or
fragments, variants, or derivatives of such peptide sequences that
are conserved across a plurality of proteins in the same or
different pathogen. The conserved regions from which the composite
sequence is derived may be conserved within subtypes of the same
pathogen or different pathogens. Preferred pathogens include, but
are not limited to bacteria, viruses, parasites, fungi and
viruses.
[0069] Composite antigens of the invention may also be obtained or
derived from the sequences of bacteria such as, for example,
multiple or combined epitopes of the proteins and/or polypeptides
of, for example, but not limited to Streptococcus, Pseudomonas,
Mycobacterium such as M. tuberculosis, Shigella, Campylobacter,
Salmonella, Haemophilus influenza, Chlamydophila pneumonia,
Corynebacterium diphtheriae, Clostridium tetani, Mycoplasma
pneumonia, Staphylococcus aureus, Moraxella catarrhalis, Legionella
pneumophila, Bordetella pertussis, Escherichia coli, such as E.
coli 0157, and multiple or combined epitomes of conserved regions
of any of the foregoing. Exemplary parasites from which sequences
may be obtained or derived include but are not limited to
Plasmodium such as Plasmodium falciparum and Trypanosoma. Exemplary
fungi include, but are not limited to Aspergillus fumigatus or
Aspergillus flavus. Exemplary viruses include, but are not limited
to arena viruses, bunyaviruses, coronaviruses, filoviruses, hepadna
viruses, herpes viruses, orthomyxoviruses, parvoviruses,
picornaviruses, papillomaviruses, reoviruses, retroviruses,
rhabdoviruses, and togaviruses. Preferably, the virus epitopes are
obtained or derived from sequences of Influenza viruses (e.g., the
paramyxoviruses).
[0070] In another preferred embodiment, the composite antigens
contain a conserved region derived from an influenza virus subtypes
(e.g., influenza viruses with varying HA or NA compositions, such
as H1N1, H5N1, H3N2, and H2N2). Epitopes of conserved regions on NA
or HA may also confer cross-subtype immunity. As an example,
conserved epitopes on NA(N1) may confer enhanced immunity to H5N1
and H1N1. With respect to similar or homologous chemical compounds
among influenza A subtypes and/or strains within a subtype,
preferably these are at least about 80 percent, more preferably at
least about 90 percent, more preferably at least about 95 percent
identical, more preferably at least about 96 percent identical,
more preferably at least about 97 percent identical, more
preferably at least about 98 percent identical, more preferably at
least about 99 percent identical, and even more preferably 100
percent identical (invariant). Preferably, at least one peptide
sequence within the composite antigen is also conserved on
homologous proteins (e.g., protein subunits) of at least two viral
particles, preferably influenza particles. Proteins of influenza
virus include, for example, expressed proteins in the virus
structure, such as HA, NA, protein polymerases (PB1, PB2, PA),
matrix proteins (M1, M2), and nucleoprotein ("NP"). Preferably, the
conserved peptide sequences are conserved on at least two or more
of the M1, M2, HA, NA, or one or more polymerase proteins.
[0071] In a preferred example, a selected sequence in the M1 and M2
proteins of the H5N1 influenza virus corresponds to the M1 and M2
proteins found in other H5N1 particles, and to the same sequence in
the M1 and M2 proteins of the H3N2 influenza virus. In addition,
while HA and NA proteins have highly variable regions, conserved
sequences from HA and NA are found across many influenza strains
and many subtypes (e.g., HA and NA sequences are conserved across
H5N1 and H1N1). In a preferred embodiment of the invention, the
composite sequences is derived from a conserved sequence present
within variants or strains (viral isolates expressing substantially
the same HA and NA proteins, but wherein the HA and NA protein
amino acid sequences show some minor drift), of a single
influenzavirus subtype and more preferably across at least two
influenzavirus subtypes, e.g., subtypes of influenza A virus.
[0072] In another embodiment, the invention provides a composite
peptide or polypeptide that includes at least one epitopic antigen,
which comprises one or more repeatedly occurring epitope sequences,
each of which is conserved across a plurality of homologous
proteins that is conserved in a population of influenzavirus
strains or serotypes, and a pharmaceutically acceptable carrier. In
exemplary composite antigens, at least one epitopic sequence is
repeated at least once, preferably at least twice times, more
preferably at least three times. In other embodiments, the at least
one epitopic sequence is repeated four or more times. Preferably,
the composite sequences are identical with the sequences in the
homologous protein subunits of at least two circulating viral
isolates. In each embodiment, the compositions may include a
pharmaceutically acceptable carrier.
[0073] In additional preferred embodiments, the composite peptide
sequences include sequences derived from genome (i.e., RNA) segment
7 of the influenza virus, while in a more preferred embodiment, the
sequences include at least portions of the M1 and M2 proteins. In
other preferred embodiments, the composite sequences include
sequences expressed from genome segments encoding the HA or NA
proteins. Such sequences are less affected by subtype drift and
more broadly protective against infections.
[0074] In additional embodiments, the composite antigen includes
one or more T-cell stimulating epitopes, such as diphtheria toxoid,
tetanus toxoid, a polysaccharide, a lipoprotein, or a derivative or
any combination thereof (including fragments or variants thereof).
Typically, the at least one repeated sequence of the composite
antigen is contained within the same molecule as the T-cell
stimulating epitopes. In the case of protein-based T-cell
stimulating epitopes, the at least one repeated sequence of the
composite antigen may be contained within the same polypeptide as
the T-cell stimulating epitopes, may be conjugated thereto, or may
be associated in other ways. Preferably, at least one repeated
sequence is incorporated within or alongside the one or more T-cell
stimulating epitopes in a composite antigen of the invention.
[0075] In additional embodiments, the composite antigens, with or
without associated T-cell stimulating epitopes may include one or
more polysaccharides or portions thereof. In preferred embodiments,
at least one sequence of a composite antigen is conjugated to one
or more polysaccharides. In other embodiments, one or more
polysaccharides are conjugated to other portions of the composite
antigen. Certain embodiments of the present invention are selected
from polysaccharide vaccines, protein-polysaccharide conjugate
vaccines, or combinations thereof.
[0076] Composite antigens of the invention may be synthesizing by
in vitro chemical synthesis, solid-phase protein synthesis, and in
vitro (cell-free) protein translation, or recombinantly engineered
and expressed in bacterial cells, fungi, insect cells, mammalian
cells, virus particles, yeast, and the like.
[0077] A preferred composite antigen includes at least one of the
following elements: at least one repeated composite sequence; at
least one T-cell epitope; at least one polysaccharide; at least one
polynucleotide; at least one structural component; or a combination
thereof. The at least one structural component may include one or
more of: at least one linker segment; at least one sugar-binding
moiety; at least one nucleotide-binding moiety; at least one
protein-binding moiety; at least one enzymatic moiety; or a
combination thereof. The invention encompasses methods of preparing
an immunogenic composition, preferably a pharmaceutical
composition, more preferably a vaccine, wherein a target antigen of
the present invention is associated with a pharmaceutically
acceptable diluent, excipient, or carrier, and may be used with
most any adjuvant.
[0078] Within the context of the present invention, that a
relatively small number of conservative or neutral substitutions
(e.g., 1 or 2) may be made within the sequence of the composite
antigen or epitope sequences disclosed herein, without
substantially altering the immunological response to the peptide.
In some cases, the substitution of one or more amino acids in a
particular peptide may in fact serve to enhance or otherwise
improve the ability of the peptide to elicit an immune or T-cell
response in an animal that has been provided with a composition
that comprises the modified peptide, or a polynucleotide that
encodes the peptide. Suitable substitutions may generally be
identified using computer programs and the effect of such
substitutions may be confirmed based on the reactivity of the
modified peptide with antisera and/or T-cells. Accordingly, within
certain preferred embodiments, a peptide for use in the disclosed
diagnostic and therapeutic methods may comprise a primary amino
acid sequence in which one or more amino acid residues are
substituted by one or more replacement amino acids, such that the
ability of the modified peptide to react with antigen-specific
antisera and/or T-cell lines or clones is not significantly less
than that for the unmodified peptide.
[0079] As described above, preferred peptide variants are those
that contain one or more conservative substitutions. A
"conservative substitution" is one in which an amino acid is
substituted for another amino acid that has similar properties,
such that one skilled in the art of peptide chemistry would expect
the secondary structure and hydropathic nature of the peptide to be
substantially unchanged. Amino acid substitutions may generally be
made on the basis of similarity in polarity, charge, solubility,
hydrophobicity, hydrophilicity and/or the amphipathic nature of the
residues. For example, negatively charged amino acids include
aspartic acid and glutamic acid; positively charged amino acids
include lysine and arginine; and amino acids with uncharged polar
head groups having similar hydrophilicity values include leucine,
isoleucine and valine; glycine and alanine; asparagine and
glutamine; and serine, threonine, phenylalanine and tyrosine.
Examples of amino acid substitutions that represent a conservative
change include: (1) replacement of one or more Ala, Pro, Gly, Glu,
Asp, Gln, Asn, Ser, or Thr; residues with one or more residues from
the same group; (2) replacement of one or more Cys, Ser, Tyr, or
Thr residues with one or more residues from the same group; (3)
replacement of one or more Val, Ile, Leu, Met, Ala, or Phe residues
with one or more residues from the same group; (4) replacement of
one or more Lys, Arg, or His residues with one or more residues
from the same group; and (5) replacement of one or more Phe, Tyr,
Trp, or His residues with one or more residues from the same group.
A variant may also, or alternatively, contain non-conservative
changes, for example, by substituting one of the amino acid
residues from group (1) with an amino acid residue from group (2),
group (3), group (4), or group (5). Variants may also (or
alternatively) be modified by, for example, the deletion or
addition of amino acids that have minimal influence on the
immunogenicity, secondary structure and hydropathic nature of the
peptide.
[0080] Epitopes may be arranged in any order relative to one
another in the composite sequence. The number of spacer amino acids
between two or more of the epitopic sequences can be of any
practical range, including, for example, from 1 or 2 amino acids to
3, 4, 5, 6, 7, 8, 9, or even 10 or more amino acids between
adjacent epitopes.
[0081] Another embodiment of the invention is directed to
polynucleotides including DNA, RNA and PNA constructs that encode
the composite sequences of the invention. These polynucleotides may
be single-stranded (coding or antisense) or double-stranded, and
may be DNA (genomic, cDNA or synthetic) or RNA molecules.
Additional coding or non-coding sequences may, but need not, be
present within a polynucleotide of the present invention, and a
polynucleotide may, but need not, be linked to other molecules
and/or support materials. As is appreciated by those of ordinary
skill in the art that, as a result of the degeneracy of the genetic
code, there are many nucleotide sequences that encode a given
primary amino acid sequence. Some of these polynucleotides bear
minimal homology to the nucleotide sequence of any native gene.
Nonetheless, polynucleotides that vary due to differences in codon
usage are specifically contemplated by the present invention.
Polynucleotides that encode an immunogenic peptide may generally be
used for production of the peptide, in vitro or in vivo. Any
polynucleotide may be further modified to increase stability in
vivo. Possible modifications include, but are not limited to, the
addition of flanking sequences at the 5' and/or 3'-ends; the use of
phosphorothioate or 2'-o-methyl rather than phosphodiesterase
linkages in the backbone; and/or the inclusion of nontraditional
bases such as inosine, queosine and wybutosine, as well as acetyl-
methyl-, thio- and other modified forms of adenine, cytidine,
guanine, thymine and uridine.
[0082] Another embodiment of the invention encompasses methods of
vaccinating a subject against Influenza that includes administering
to a patient in need of influenza vaccination a therapeutically or
prophylactically effective amount of an influenza vaccine, which
influenza vaccine includes a composite antigen comprising one or
more repeatedly occurring composite or other sequences, each of
which is conserved across a plurality of homologous proteins in a
plurality of influenza virus particles, and a pharmaceutically
acceptable carrier, to provide a detectable immune response in the
patient against influenza.
[0083] Another embodiment of the invention is directed to
nucleotide or DNA vaccines encoding composite antigens of the
invention. A DNA vaccine of the invention contains the genetic
sequence of a composite antigen, plus other necessary sequences
that provide for the expression of the composite antigen in cells.
By injecting the mammal with the genetically engineered DNA, the
composite antigen is produced in or preferably on cells, which the
mammal's immune system recognizes and thereby generates a humoral
or cellular response to the composite antigen, and therefore the
pathogen. DNA vaccines have a number of advantages over
conventional vaccines, including the ability to induce a more
general and complete immune response in the mammal. Accordingly,
DNA vaccines can be used to protect a mammal against disease caused
from many different pathogenic organisms of viral, bacterial, and
parasitic origin as well as certain tumors.
[0084] DNA vaccines typically comprise a bacterial DNA contained
that encodes the composite antigen contained in vectors or plasmids
that have been genetically modified to transcribe and translate the
composite antigenic sequences into specific protein sequences
derived from a pathogen. By way of example, the vaccine DNA is
injected into the cells of the body, where the cellular machinery
transcribed and translates the DNA into the composite antigen.
Composite antigens, being non-natural and unrecognized by the
mammalian immune system, are processed by cells and the processed
proteins, preferably the epitopes, displayed on cell surfaces. Upon
recognition of these composite antigens as foreign, the mammal's
immune system generates an appropriate immune response that
protects the mammal from infection. In addition, DNA vaccine of the
invention are preferably codon optimized for expression in the
mammalian cells of interest, such as but not limited to mouse or
human cells. In a preferred embodiment, codon optimization involves
selecting a desired codon usage bias (the frequency of occurrence
of synonymous codons in coding DNA) for the particular cell type so
that the desired peptide sequence is expressed.
[0085] Another embodiment of the invention is directed to
therapeutic and prophylactic agents in a pharmaceutically
acceptable composition for administration to a cell or an animal,
either alone, or in combination with one or more other modalities
of prophylaxis and/or therapy. Therapeutic and prophylactic agents
of the invention include composite antigens, composite epitopes,
compositions containing composite antigens and epitopes, composite
sequences, DNA vaccines of the invention, antibodies of the
invention, and/or T cells primed or exposed to composite antigens
of the invention. The formulation of pharmaceutically-acceptable
excipients and carrier solutions is well known to those of ordinary
skill in the art, as is the development of suitable dosing and
treatment regimens for using the particular compositions described
herein in a variety of treatment regimens.
[0086] The amount of immunogenic composition(s) and the time needed
for the administration of such immunogenic composition(s) will be
within the purview of the ordinary-skilled artisan having benefit
of the present teachings. The administration of a
therapeutically-effective, pharmaceutically-effective, and/or
prophylactically-effective amount of the disclosed immunogenic
compositions may be achieved by a single administration, such as
for example, a single injection of a sufficient quantity of the
delivered agent to provide the desired benefit to the patient
undergoing such a procedure. Alternatively, in some circumstances,
it may be desirable to provide multiple, or successive
administrations of the immunogenic compositions, either over a
relatively short, or even a relatively prolonged period of time, as
may be determined by the medical practitioner overseeing the
administration of such compositions to the selected individual.
[0087] The immunogenic compositions and vaccines of the present
invention are preferably administered in a manner compatible with
the dosage formulation, and in such an amount as will be
prophylactically or therapeutically effective and preferably
immunogenic. The quantity to be administered depends on the subject
to be treated, including, e.g., the capacity of the patient's
immune system to mount an immune response, and the degree of
protection desired. Suitable dosage ranges may be on the order of
several hundred micrograms (.mu.g) of active ingredient per
vaccination with a preferred range from about 0.1 .mu.g to 2000
.mu.g (even though higher amounts, such as, e.g., in the range of
about 1 to about 10 mg are also contemplated), such as in the range
from about 0.5 .mu.g to 1000 .mu.g, preferably in the range from
about 1 .mu.g to about 500 .mu.g and especially in the range from
about 10 .mu.g to about 100 .mu.g. Suitable regimens for initial
administration and booster shots are also variable but are typified
by an initial administration followed by optional but preferred
subsequent inoculations or other periodic administrations.
[0088] In certain embodiments, the dose would consist of the range
of about 1 .mu.g to about 1 mg total protein or target antigen. In
one exemplary embodiment, the vaccine dosage range is about 0.1
.mu.g to about 10 mg. However, one may prefer to adjust dosage
based on the amount of peptide delivered. In either case, these
ranges are merely guidelines from which one of ordinary skill in
the art may deviate according to conventional dosing techniques.
Precise dosages may be determined by assessing the immunogenicity
of the conjugate produced in the appropriate host so that an
immunologically effective dose is delivered. An immunologically
effective dose is one that stimulates the immune system of the
patient to establish an immune response to the immunogenic
composition or vaccine. Preferably, a level of immunological memory
sufficient to provide long-term protection against disease caused
by microbial infection is obtained. The immunogenic compositions or
vaccines of the invention may be preferably formulated with an
adjuvant. By "long-term" it is preferably meant over a period of
time of at least about 6 months, over at least about 1 year, over
at least about 2 to 5 or even at least about 2 to about 10 years or
longer.
[0089] Another embodiment of the invention is directed to
antibodies that are specific for the composite antigens as
described here and conservative variants thereof. Antibodies
specific for these polypeptides are useful, e.g., in both
diagnostic and therapeutic purposes, e.g., related to the activity,
distribution, and expression of target polypeptides. Antibodies of
the invention may be classes IgG, IgM, IgA, IgD or IgE and include,
but are not limited to, polyclonal antibodies, monoclonal
antibodies, multiple or single chain antibodies, including single
chain Fv (sFv or scFv) antibodies in which a variable heavy and a
variable light chain are joined together (directly or through a
peptide linker) to form a continuous polypeptide, and humanized or
chimeric antibodies.
[0090] Antibodies specific for the composite peptides of the
invention can be generated by methods well known in the art. Such
antibodies can include, but are not limited to, polyclonal,
monoclonal, chimeric, humanized, single chain, Fab fragments and
fragments produced by an Fab expression library. Numerous methods
for producing polyclonal and monoclonal antibodies are known to
those of skill in the art, and can be adapted to produce antibodies
specific for the polypeptides of the invention, and/or encoded by
the polynucleotide sequences of the invention (see, e.g., Coligan
Current Protocols in Immunology Wiley/Greene, NY; Paul (ed.)
(1991); (1998) Fundamental Immunology Fourth Edition,
Lippincott-Raven, Lippincott Williams & Wilkins; Harlow and
Lane (1989) Antibodies: A Laboratory Manual, Cold Spring Harbor
Press, NY, USA; Stites et al. (Eds.) Basic and Clinical Immunology
(4th ed.) Lange Medical Publications, Los Altos, Calif., USA and
references cited therein; Goding, Monoclonal Antibodies: Principles
and Practice (2d ed.) Academic Press, New York, N.Y., USA; 1986;
and Kohler and Milstein (1975).
[0091] The following examples illustrate embodiments of the
invention, but should not be viewed as limiting the scope of the
invention.
EXAMPLES
Sequences
[0092] The following is a list of exemplary sequence. These
sequences include composite sequences as well as sequences of
interest that can be combined to form composite sequences of the
invention:
TABLE-US-00002 SEQ ID NO 1 DWSGYSGSFVQHPELTGLD (N1 sequence; H1 N5
SEQ ID NO 2 ETPIRNE (N1 epitope) SEQ ID NO 3 FVIREPFISCSHLEC (Pep
5) SEQ ID NO 4 GNFIAP (HA epitope; Pep 1) SEQ ID NO 5 GNLIAP (HA
epitope; Pep 2) SEQ ID NO 6 GNLFIAP (composite sequence of SEQ ID
NOs 4 and 5; Pep 3) SEQ ID NO 7 GNLIFAP (composite sequence of SEQ
ID NOs 4 and 5) SEQ ID NO 8 HYEECSCY (NA epitope; Pep 10) SEQ ID NO
9 LLTEVETPIR SEQ ID NO 10 LLTEVETPIRN SEQ ID NO 11 LLTEVETPIRNE SEQ
ID NO 12 DWSGYSGSFVQHPELTGL (N1 sequence; H1 N5) SEQ ID NO 13
EVETPIRNE SEQ ID NO 14 FLLPEDETPIRNEWGLLTDDETPIRYIKANSKFIGITE SEQ
ID NO 15
GNLFIAPGNLFIAPHYEECSCYHYEECSCYQYIKANSKFIGITEHYEECSCYTPIRNETPIRNE
SEQ ID NO 16 GNLFIAPGNLFIAPQYIKANSKFIGITEGNLFIAP (composite of SEQ
ID NO 6, SEQ ID NO 6, SEQ ID NO 60, and SEQ ID NO 6) SEQ ID NO 17
HYEECSCYDWSGYSGSFVQHPELTGLHYEECSCYQYIKANSKFIGITE SEQ ID NO 18
ITGFAPFSKDNSIRLSAGGDIWVTREPYVSCDP SEQ ID NO 19 IWGIHHP (HA epitope)
SEQ ID NO 20 IWGVHHP (HA epitope) SEQ ID NO 21 IWGVIHHP (composite
of SEQ ID NOs. 19 and 20) SEQ ID NO 22 IWGIVHHP (composite of SEQ
ID NOs. 19 and 20) SEQ ID NO 23 KSCINRCFYVELIRGR SEQ ID NO 24
LLTEVETPIRNESLLTEVETPIRNEWG (M2e epitope) SEQ ID NO 25
LLTEVETPIRNEW (M2e epitope) SEQ ID NO 26 LLTEVETPIRNEWG (M2e
epitope) SEQ ID NO 27 LTEVETPIRNE (M2e epitope) SEQ ID NO 28
LTEVETPIRNEW (M2e epitope) SEQ ID NO 29 LTEVETPIRNEWG (M2e epitope)
SEQ ID NO 30 MSLLTEVET (M2e epitope) SEQ ID NO 31 MSLLTEVETP (M2e
epitope) SEQ ID NO 32 MSLLTEVETPI (M2e epitope) SEQ ID NO 33
MSLLTEVETPIR (M2e epitope) SEQ ID NO 34 MSLLTEVETPIRN (M2e epitope)
SEQ ID NO 35 MSLLTEVETPIRNE (M2e epitopes) SEQ ID NO 36
MSLLTEVETPIRNETPIRNE (M2e epitope) SEQ ID NO 37 MSLLTEVETPIRNEW
(M2e epitope) SEQ ID NO 38 MSLLTEVETPIRNEWG (M2e epitope) SEQ ID NO
39 MSLLTEVETPIRNEWGCRCNDSSD (M2e epitope) SEQ ID NO 40 SLLTEVET
(M2e epitope) SEQ ID NO 41 SLLTEVETPIRNE (M2e epitope) SEQ ID NO 42
SLLTEVETPIRNEW (M2e epitope) SEQ ID NO 43 SLLTEVETPIRNEWG (M2e
epitope) SEQ ID NO 44 SLLTEVETPIRNEWGTPIRNE (M2e epitope) SEQ ID NO
45 SLLTEVETPIRNEWGTPIRNETPIRNE (M2e epitope) SEQ ID NO 46
SLLTEVETPIRNEWGTPIRNETPIRNETPIRNE (M2e epitopes) SEQ ID NO 47
SLLTEVETPIRNEWGLLTEVETPIRQYIKANSKFIGITE (M2e epitope) SEQ ID NO 48
TEVETPIRNE (M2e epitope) SEQ ID NO 49 TPIRNE SEQ ID NO 50 VETPIRNE
SEQ ID NO 51 VTREPYVSCDPKSCINRCFYVELIRGRVTREPYVSCDPWYIKANSKFIGITE
SEQ ID NO 52 WGIHHP (HA conserved region; Pep 5) SEQ ID NO 53
WGVHHP (HA conserved region; Pep 4) SEQ ID NO 54 WGVIHHP (composite
of SEQ ID NOs 52 and 53; Pep 6) SEQ ID NO 55 WGIVHHP (composite of
SEQ ID NOs 52 and 53; Pep 7) SEQ ID NO 56 YIWGIHHP SEQ ID NO 57
YIWGVHHP SEQ ID NO 58 YIWGVIHHP (composite of SEQ ID NOs 56 and 57)
SEQ ID NO 59 YIWGIVHHP (composite of SEQ ID NOs 56 and 57) SEQ ID
NO 60 QYIKANSKFIGITE SEQ ID NO 61 PIRNEWGCRCNDSSD SEQ ID NO 65
SEYAYGSFVRTVSLPVGADEGNLFIAPWGVIHHPHYEECSCYGLPVEYLQVPSPSMGRDI
KVQFQSGGANSPALYLLDGLRAQDDFSGWDINTPAFEWYDQSGLSVVMPVGGQSSFYS
DWYQPACRKAGCQTYKWETFLTSELPGWLQANRHVQPTGSAVVGLSMAASSALTLAI
YHPQQFVYAGAMSGLLDPSQAMGPTLIGLAMGDAGGYKASDMWGPKEDPAWQRNDP
LLNVGKLIANNTRVWVYCGNGKPSDLGGNNLPAKFLEGFVRTSNIKFQDAYNAGGHNG
VFDFPDSGTHSWEYWGAQLNAMKPDLQRHWVPRPTPGPPQGAFDFPDSGTHSWEYWG
AQLNAMKPDLQRHWVPRPTPGPPQGA (Sequence for DNA vaccine development
373 amino acids; has a TB conserved regions on each side of Pep 11)
SEQ ID NO 66 GNLFIAPWGVIHHPHYEECSCY (SEQ ID NOs 6,54, and 8; Pep
11) SEQ ID NO 67 WGVIHHPGNLFIAPHYEECSCY (SEQ ID NOs 54,6, and 8)
SEQ ID NO 68
SRPGLPVEYLQVPSPSMGRDIKVQFQSGGANSPALYLLDGLRAQDDFSGWDINTPAFEW
YDQSGLSVVMPVGGQSSFYSDWYQPACGKAGCQTYKWETFLTSELPGWLQANRHVKP
TGSAVVGLSMAASSALTLAIYHPQQFVYAGAMSGLLDPSQAMGPTLIGLAMGDAGGY
KASDMWGPKEDPAWQRNDPLLNVGKLIANNTRVWVYCGNGKPSDLGGNNLPAKFLE
GFVRTSNIKFQDAYNAGGGHNGVFDFPDSGTHSWEYWGAQLNAMKPDLQRALGATPN TGPAPQGA
(TB coding region sequence of 85a) SEQ ID NO 69
TCCCGGCCGGGCTTGCCGGTGGAGTACCTGCAGGTGCCGTCGCCGTCGATGGGCCGT
GACATCAAGGTCCAATTCCAAAGTGGTGGTGCCAACTCGCCCGCCCTGTACCTGCTC
GACGGCCTGCGCGCGCAGGACGACTTCAGCGGCTGGGACATCAACACCCCGGCGTT
CGAGTGGTACGACCAGTCGGGCCTGTCGGTGGTCATGCCGGTGGGTGGCCAGTCAA
GCTTCTACTCCGACTGGTACCAGCCCGCCTGCGGCAAGGCCGGTTGCCAGACTTACA
AGTGGGAGACCTTCCTGACCAGCGAGCTGCCGGGGTGGCTGCAGGCCAACAGGCAC
GTCAAGCCCACCGGAAGCGCCGTCGTCGGTCTTTCGATGGCTGCTTCTTCGGCGCTG
ACGCTGGCGATCTATCACCCCCAGCAGTTCGTCTACGCGGGAGCGATGTCGGGCCTG
TTGGACCCCTCCCAGGCGATGGGTCCCACCCTGATCGGCCTGGCGATGGGTGACGCT
GGCGGCTACAAGGCCTCCGACATGTGGGGCCCGAAGGAGGACCCGGCGTGGCAGCG
CAACGACCCGCTGTTGAACGTCGGGAAGCTGATCGCCAACAACACCCGCGTCTGGG
TGTACTGCGGCAACGGCAAGCCGTCGGATCTGGGTGGCAACAACCTGCCGGCCAAG
TTCCTCGAGGGCTTCGTGCGGACCAGCAACATCAAGTTCCAAGACGCCTACAACGCC
GGTGGCGGCCACAACGGCGTGTTCGACTTCCCGGACAGCGGTACGCACAGCTGGGA
GTACTGGGGCGCGCAGCTCAACGCTATGAAGCCCGACCTGCAACGGGCACTGGGTG
CCACGCCCAACACCGGGCCCGCGCCCCAGGGCGCC (nucleotide sequence
corresponding to TB sequence 85a or SEQ ID NO 64) SEQ ID NO 70
SEFAYGSFVRTVSLPVGADEGNLFIAPWGVIHHPHYEECSCYSRPGLPVEYLQVPSPSMG
RDIKVQFQSGGANSPALYLLDGLRAQDDFSGWDINTPAFEWYDQSGLSVVMPVGGQSS
FYSDWYQPACGKAGCQTYKWETFLTSELPGWLQANRHVKPTGSAVVGLSMAASSALT
LAIYHPQQFVYAGAMSGLLDPSQAMGPTLIGLAMGDAGGYKASDMWGPKEDPAWQR
NDPLLNVGKLIANNTRVWVYCGNGKPSDLGGNNLPAKFLEGFVRTSNIKFQDAYNAGG
GHNGVFDFPDSGTHSWEYWGAQLNAMKPDLQRALGATPNTGPAPQGA (336 amino acid
sequence comprising HSPx, Pep 11 and TB 85a) SEQ ID NO 71
TTTGGGCCCATTATGTCGGAATTCGCGTACGGTTCCTTCGTTCGCACGGTGTCGCTGC
CGGTAGGTGCTGACGAGGGGAATCTAttcATTGCTCCTTGGGGGGTTattCA
CCACCCGCATTATGAGGAATGTTCCTGTTACTCCCGGCCGGGCTTGCCGG
TGGAGTACCTGCAGGTGCCGTCGCCGTCGATGGGCCGTGACATCAAGGTCCAATTCC
AAAGTGGTGGTGCCAACTCGCCCGCCCTGTACCTGCTCGACGGCCTGCGCGCGCAG
GACGACTTCAGCGGCTGGGACATCAACACCCCGGCGTTCGAGTGGTACGACCAGTC
GGGCCTGTCGGTGGTCATGCCGGTGGGTGGCCAGTCAAGCTTCTACTCCGACTGGTA
CCAGCCCGCCTGCGGCAAGGCCGGTTGCCAGACTTACAAGTGGGAGACCTTCCTGA
CCAGCGAGCTGCCGGGGTGGCTGCAGGCCAACAGGCACGTCAAGCCCACCGGAAGC
GCCGTCGTCGGTCTTTCGATGGCTGCTTCTTCGGCGCTGACGCTGGCGATCTATCACC
CCCAGCAGTTCGTCTACGCGGGAGCGATGTCGGGCCTGTTGGACCCCTCCCAGGCGA
TGGGTCCCACCCTGATCGGCCTGGCGATGGGTGACGCTGGCGGCTACAAGGCCTCC
GACATGTGGGGCCCGAAGGAGGACCCGGCGTGGCAGCGCAACGACCCGCTGTTGAA
CGTCGGGAAGCTGATCGCCAACAACACCCGCGTCTGGGTGTACTGCGGCAACGGCA
AGCCGTCGGATCTGGGTGGCAACAACCTGCCGGCCAAGTTCCTCGAGGGCTTCGTGC
GGACCAGCAACATCAAGTTCCAAGACGCCTACAACGCCGGTGGCGGCCACAACGGC
GTGTTCGACTTCCCGGACAGCGGTACGCACAGCTGGGAGTACTGGGGCGCGCAGCT
CAACGCTATGAAGCCCGACCTGCAACGGGCACTGGGTGCCACGCCCAACACCGGGC
CCGCGCCCCAGGGCGCCTAGTTTCTTAAGTTT Underlined sequences: Start and
stop codons Afl II Restriction Site (NEB Buffer 4) Note: multi T
Spacer between stop and Afl II RE Apa I Restriction Site (NEB
Buffer 4) Note: Spacer between start and Apa I RE is Kozak minimal
translation initiation site Bold sequences: HA 1 sequence bolded
and underlined (SEQ ID NO 72) HA2 sequence bolded (SEQ ID NO 73 NA1
sequence bolded and underlined (SEQ ID NO 74) SEQ ID NO 72
GGGAATCTAttcATTGCTCCT SEQ ID NO 73 TGGGGGGTTattCACCACCCG SEQ ID NO
74 CATTATGAGGAATGTTCCTGTTAC SEQ ID NO 75 SEFAYGSFVRTVSLPVGADE (Heat
Shock protein sequence of epitope HSPx derived from Mycobacterium
tuberculosis H37Rv (NC_000962.2) SEQ ID NO 76
TCGGAATTCGCGTACGGTTCCTTCGTTCGCACGGTGTCGCTGC CGGTAGGTGCTGACGAG
(nucleotide sequence corresponding to HSPx; SEQ ID NO 66) SEQ ID NO
77 GNLFIAPWGVIHHPHYEECSCY (underlined sequences are epitopes HA
{composite} and NA, respectively, of Influenza A; Pep 11) SEQ ID NO
78 GGGAATCTAttcATTGCTCCTTGGGGGGTTattCACCACCCG
CATTATGAGGAATGTTCCTGTTAC (Pep 11 - SEQ ID NO 68) SEQ ID NO 79
TCGGAATTCGCGTACGGTTCCTTCGTTCGCACGGTGTCGCTGCCGGTAGGTGCTGAC
GAGGGGAATCTAttcATTGCTCCTTGGGGGGTTattCACCACCCGCATTATGAGGAATGT
TCCTGTTACTCCCGGCCGGGCTTGCCGGTGGAGTACCTGCAGGTGCCGTCGCCGTCG
ATGGGCCGTGACATCAAGGTCCAATTCCAAAGTGGTGGTGCCAACTCGCCCGCCCTG
TACCTGCTCGACGGCCTGCGCGCGCAGGACGACTTCAGCGGCTGGGACATCAACAC
CCCGGCGTTCGAGTGGTACGACCAGTCGGGCCTGTCGGTGGTCATGCCGGTGGGTGG
CCAGTCAAGCTTCTACTCCGACTGGTACCAGCCCGCCTGCGGCAAGGCCGGTTGCCA
GACTTACAAGTGGGAGACCTTCCTGACCAGCGAGCTGCCGGGGTGGCTGCAGGCCA
ACAGGCACGTCAAGCCCACCGGAAGCGCCGTCGTCGGTCTTTCGATGGCTGCTTCTT
CGGCGCTGACGCTGGCGATCTATCACCCCCAGCAGTTCGTCTACGCGGGAGCGATGT
CGGGCCTGTTGGACCCCTCCCAGGCGATGGGTCCCACCCTGATCGGCCTGGCGATGG
GTGACGCTGGCGGCTACAAGGCCTCCGACATGTGGGGCCCGAAGGAGGACCCGGCG
TGGCAGCGCAACGACCCGCTGTTGAACGTCGGGAAGCTGATCGCCAACAACACCCG
CGTCTGGGTGTACTGCGGCAACGGCAAGCCGTCGGATCTGGGTGGCAACAACCTGC
CGGCCAAGTTCCTCGAGGGCTTCGTGCGGACCAGCAACATCAAGTTCCAAGACGCC
TACAACGCCGGTGGCGGCCACAACGGCGTGTTCGACTTCCCGGACAGCGGTACGCA
CAGCTGGGAGTACTGGGGCGCGCAGCTCAACGCTATGAAGCCCGACCTGCAACGGG
CACTGGGTGCCACGCCCAACACCGGGCCCGCGCCCCAGGGCGCC (1008 nucleotide DNA
construct of composite peptide comprising TB epitopes at each end
with an influenza sequence in the middle which is composed of three
epitopes - 2 HA composites {underlined} with an NA epitope between
them). SEQ ID NO 80 CAGAGNFIAP SEQ ID NO 81 CAGAGNLIAP SEQ ID NO 82
CAGAGNLFIAP SEQ ID NO 83 CAGAWGVHHP SEQ ID NO 84 CAGAWGIHHP SEQ ID
NO 85 CAGAWGVIHHP SEQ ID NO 86 CAGAWGIVHHP SEQ ID NO 87
GNLIAPWGVIHHP SEQ ID NO 88 CAGAGNLIAPWGVIHHP SEQ ID NO 89
GNLFIAPWGVIHHP SEQ ID NO 90 CAGAGNLFIAPWGVIHHP SEQ ID NO 91
HYEECSCY SEQ ID NO 92 CAGAHYEECSCY SEQ ID NO 93
GNLFIAPWGVIHHPHYEECSCY SEQ ID NO 94 CAGAGNLFIAPWGVIHHPHYEECSCY SEQ
ID NO 95 GNLFIAPWGVIHHPGNLFIAPWGVIHHP SEQ ID NO 96
CAGAGNLFIAPWGVIHHPGNLFIAPWGVIHHP SEQ ID NO 97
HYEECSCYGNLFIAPWGVIHHP SEQ ID NO 98 GNLFIAPHYEECSCYWGVIHHP SEQ ID
NO 99 SLLTEVETPIRNEWGLLTEVETPIRQYIKANSKFIGITE SEQ ID NO 100
GNLFIAPGNLFIAPQYIKANSKFIGITEGNLFIAP SEQ ID NO 101
HYEECSCYDWSGYSGSFVQHPELTGLHYEECSCYQYIKANSKFIGITE SEQ ID NO 102
VTREPYVSCDPKSCINRCFYVELIRGRVTREPYVSCDPQYIKANSKFIGITE SEQ ID NO 103
DWSGYSGSFVQHPELTGL SEQ ID NO 104 ITGFAPFSKDNSIRLSAGGDIWVTREPYVSCDP
SEQ ID NO 105 KSCINRCFYVELIRGR SEQ ID NO 106 GNLFIAPRYAFA SEQ ID NO
107 CAGAGNLFIAPRYAFA SEQ ID NO 108 GNLVVPRYAFA SEQ ID NO 109
CAGAGNLVVPRYAFA SEQ ID NO 110 GNLIAPRYAFA SEQ ID NO 111
CAGAGNLIAPRYAFA SEQ ID NO 112 GNLVVP SEQ ID NO 113 CAGAGNLVVP
SEQ ID NO 114 FVIREPFISCSHLEC SEQ ID NO 115 CAGAFVIREPFISCSHLEC
[0093] FIG. 1 shows titers as determined by ELISA of mice
vaccinated with Pep 6, Pep 9, Pep 10 or Pep 11. As can be seen,
vaccinations with Pep 9, Pep 10 and Pep 11 provided a generally
strong repose to the native antigen and the highest titers in
mice.
[0094] FIG. 2 shows mean OD values of sera from mice immunized
peptides derived from the Wuhan strain of Influenza (H3N2). Results
indicate that Pep 10 and Pep 11 provide a significant immune
response as compared to unvaccinated mice (the Balb/c pool) and
mice vaccinated with Pep 3.
[0095] FIG. 3 shows mean OD values of antisera (titered at 1:100)
following immunization with Pep 6. As can be seen, Pep 6 the
antisera reacted strongly with viruses of both H1N1 and H3N2.
[0096] FIG. 4 shows antisera titers of mice immunized with Pep 9 as
reacted with Influenza strain Caledonia virus (H1N1) at various
dilutions. As seen, Pep 9 reacts strongly with whole virus.
[0097] FIG. 5 shows four graphs (A-D), each depicting the
absorbance of Pep 9 sera to one of four substrates: (A) fresh Wuhan
virus; (B) fixed Wuhan virus; (C) fresh Caledonia virus; and (D)
fixed Caledonia virus. As shown, Pep 9 sera reacting strongly in
with all substrates tested.
[0098] FIG. 6 shows the titers from mice immunized with Pep 9 as
captured with substrates of fresh or fixed Wuhan or Caledonia virus
substrates. Again, Pep 9 antisera were strongly reactive with each
and the amount of binding observed for many of the antisera tested
was similar between fresh and fixed.
[0099] FIG. 7 shows antisera titers of mice immunized with Pep 11
as captured with various dilutions of substrates of H5N1. As can be
seen, binding was observed with both sera tested.
[0100] A composite antigen that contains previously determined
conserved surface protein epitopes of hemagglutinin and
neuraminidase from Influenza A viruses (several subtypes including
human H1, H3 and high-path H5) was constructed. Also included were
proteins segments from HspX and 85a from Mycobacterium
tuberculosis. The expressed protein hybrid construct included the
following: NH2+----HspX-HA1-HA2-NA1-85a-----COO-- wherein, HspX is
20 amino acids from Mycobacterium tuberculosis HspX protein; HA1 is
a 7 amino acid highly conserved hybrid region influenza A
hemagglutinin protein; HA2 is a 7 amino acid highly conserved
hybrid region of influenza A hemagglutinin protein; NA1 is an 8
amino acid highly conserve hybrid region of influenza A
neuraminidase protein; and 85a is 294 amino acids from the 85 kD
85a protein of Mycobacterium tuberculosis. The mature corresponding
protein sequence is the 336 amino acid sequence of SEQ ID NO
70.
[0101] The composite antigen vaccine is constructed using the pVAX1
(Invitrogen Inc, Cat # V260-20) by recombinant methods. Briefly, a
single stranded (ss) DNA polymer corresponding to SEQ ID 70 is
synthesized. The ssDNA sequence also include: 1) a 5' ApaI
restriction endonuclease recognition site and a 3' Afl II
restriction site for directional insertion into pVAX1 cloning
vector, 2) a ATT minimal Kozak translation initiation sequence at
the 5' end, 3) a ATG start codon, and 4) a 3' TAG termination
(stop) codon. The nucleotide sequence in its entirety will consist
of 1038 nucleotides which is SEQ ID NO 71.
[0102] This single stranded nucleotide sequence is used as a
template to generate double stranded amplicon by polymerase chain
reaction. Following PCR amplification the amplicon is subjected to
restriction endonuclease digestion, ligated into pVAX1 and
transformed into E. coli bacteria using standard transformation and
screening methods with Kanamycin selection. Plasmids containing
recombinant insert are grown in overnight culture and purified
using known plasmid extraction methods. Concentrations of
recombinant pVAX1 plasmid are subjected to DNA sequencing and
utilized in downstream transfection experiments in mice.
[0103] FIGS. 8-1 to 8-6 list a large number of sequences that
include conserved or otherwise targeted regions of various genetic
sequence of interest, and composite sequences of the invention.
[0104] Other embodiments and uses of the invention will be apparent
to those skilled in the art from consideration of the specification
and practice of the invention disclosed herein. All references
cited herein, including all publications, U.S. and foreign patents
and patent applications, are specifically and entirely incorporated
by reference. The term comprising, where ever used, is intended to
include the terms consisting and consisting essentially of.
Furthermore, the terms comprising, including, and containing are
not intended to be limiting. It is intended that the specification
and examples be considered exemplary only with the true scope and
spirit of the invention indicated by the following claims.
Sequence CWU 1 SEQUENCE LISTING <160> NUMBER OF SEQ ID
NOS: 116 <210> SEQ ID NO 1 <211> LENGTH: 19 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 1 Asp Trp Ser Gly Tyr Ser Gly Ser Phe Val Gln His Pro Glu
Leu Thr 1 5 10 15 Gly Leu Asp <210> SEQ ID NO 2 <211>
LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 2 Glu Thr Pro Ile Arg Asn Glu 1 5
<210> SEQ ID NO 3 <211> LENGTH: 15 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 3
Phe Val Ile Arg Glu Pro Phe Ile Ser Cys Ser His Leu Glu Cys 1 5 10
15 <210> SEQ ID NO 4 <211> LENGTH: 6 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 4
Gly Asn Phe Ile Ala Pro 1 5 <210> SEQ ID NO 5 <211>
LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 5 Gly Asn Leu Ile Ala Pro 1 5
<210> SEQ ID NO 6 <211> LENGTH: 7 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 6 Gly
Asn Leu Phe Ile Ala Pro 1 5 <210> SEQ ID NO 7 <211>
LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 7 Gly Asn Leu Ile Phe Ala Pro 1 5
<210> SEQ ID NO 8 <211> LENGTH: 8 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 8 His
Tyr Glu Glu Cys Ser Cys Tyr 1 5 <210> SEQ ID NO 9 <211>
LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 9 Leu Leu Thr Glu Val Glu Thr Pro Ile
Arg 1 5 10 <210> SEQ ID NO 10 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 10 Leu Leu Thr Glu Val Glu Thr Pro Ile Arg
Asn 1 5 10 <210> SEQ ID NO 11 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 11 Leu Leu Thr Glu Val Glu Thr Pro Ile Arg
Asn Glu 1 5 10 <210> SEQ ID NO 12 <211> LENGTH: 18
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 12 Asp Trp Ser Gly Tyr Ser Gly Ser Phe Val
Gln His Pro Glu Leu Thr 1 5 10 15 Gly Leu <210> SEQ ID NO 13
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 13 Glu Val Glu Thr Pro Ile
Arg Asn Glu 1 5 <210> SEQ ID NO 14 <211> LENGTH: 38
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 14 Phe Leu Leu Pro Glu Asp Glu Thr Pro Ile
Arg Asn Glu Trp Gly Leu 1 5 10 15 Leu Thr Asp Asp Glu Thr Pro Ile
Arg Tyr Ile Lys Ala Asn Ser Lys 20 25 30 Phe Ile Gly Ile Thr Glu 35
<210> SEQ ID NO 15 <211> LENGTH: 64 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 15
Gly Asn Leu Phe Ile Ala Pro Gly Asn Leu Phe Ile Ala Pro His Tyr 1 5
10 15 Glu Glu Cys Ser Cys Tyr His Tyr Glu Glu Cys Ser Cys Tyr Gln
Tyr 20 25 30 Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu His
Tyr Glu Glu 35 40 45 Cys Ser Cys Tyr Thr Pro Ile Arg Asn Glu Thr
Pro Ile Arg Asn Glu 50 55 60 <210> SEQ ID NO 16 <211>
LENGTH: 35 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 16 Gly Asn Leu Phe Ile Ala Pro Gly Asn
Leu Phe Ile Ala Pro Gln Tyr 1 5 10 15 Ile Lys Ala Asn Ser Lys Phe
Ile Gly Ile Thr Glu Gly Asn Leu Phe 20 25 30 Ile Ala Pro 35
<210> SEQ ID NO 17 <211> LENGTH: 48 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 17
His Tyr Glu Glu Cys Ser Cys Tyr Asp Trp Ser Gly Tyr Ser Gly Ser 1 5
10 15 Phe Val Gln His Pro Glu Leu Thr Gly Leu His Tyr Glu Glu Cys
Ser 20 25 30 Cys Tyr Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile Gly
Ile Thr Glu 35 40 45 <210> SEQ ID NO 18 <211> LENGTH:
33 <212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 18 Ile Thr Gly Phe Ala Pro Phe Ser Lys Asp
Asn Ser Ile Arg Leu Ser 1 5 10 15 Ala Gly Gly Asp Ile Trp Val Thr
Arg Glu Pro Tyr Val Ser Cys Asp 20 25 30 Pro <210> SEQ ID NO
19 <211> LENGTH: 7 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 19 Ile Trp Gly Ile
His His Pro 1 5 <210> SEQ ID NO 20 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 20 Ile Trp Gly Val His His Pro 1 5
<210> SEQ ID NO 21 <211> LENGTH: 8 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 21
Ile Trp Gly Val Ile His His Pro 1 5 <210> SEQ ID NO 22
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 22 Ile Trp Gly Ile Val His
His Pro 1 5 <210> SEQ ID NO 23 <211> LENGTH: 16
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 23 Lys Ser Cys Ile Asn Arg Cys Phe Tyr Val
Glu Leu Ile Arg Gly Arg 1 5 10 15 <210> SEQ ID NO 24
<211> LENGTH: 27 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 24 Leu Leu Thr Glu Val Glu
Thr Pro Ile Arg Asn Glu Ser Leu Leu Thr 1 5 10 15 Glu Val Glu Thr
Pro Ile Arg Asn Glu Trp Gly 20 25 <210> SEQ ID NO 25
<211> LENGTH: 13 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 25 Leu Leu Thr Glu Val Glu
Thr Pro Ile Arg Asn Glu Trp 1 5 10 <210> SEQ ID NO 26
<211> LENGTH: 14 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 26 Leu Leu Thr Glu Val Glu
Thr Pro Ile Arg Asn Glu Trp Gly 1 5 10 <210> SEQ ID NO 27
<211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 27 Leu Thr Glu Val Glu Thr
Pro Ile Arg Asn Glu 1 5 10 <210> SEQ ID NO 28 <211>
LENGTH: 12 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 28 Leu Thr Glu Val Glu Thr Pro Ile Arg
Asn Glu Trp 1 5 10 <210> SEQ ID NO 29 <211> LENGTH: 13
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 29 Leu Thr Glu Val Glu Thr Pro Ile Arg Asn
Glu Trp Gly 1 5 10 <210> SEQ ID NO 30 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 30 Met Ser Leu Leu Thr Glu Val Glu Thr 1 5
<210> SEQ ID NO 31 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 31
Met Ser Leu Leu Thr Glu Val Glu Thr Pro 1 5 10 <210> SEQ ID
NO 32 <211> LENGTH: 11 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 32 Met Ser Leu Leu
Thr Glu Val Glu Thr Pro Ile 1 5 10 <210> SEQ ID NO 33
<211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 33 Met Ser Leu Leu Thr Glu
Val Glu Thr Pro Ile Arg 1 5 10 <210> SEQ ID NO 34 <211>
LENGTH: 13 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 34 Met Ser Leu Leu Thr Glu Val Glu Thr
Pro Ile Arg Asn 1 5 10 <210> SEQ ID NO 35 <211> LENGTH:
14 <212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 35 Met Ser Leu Leu Thr Glu Val Glu Thr Pro
Ile Arg Asn Glu 1 5 10 <210> SEQ ID NO 36 <211> LENGTH:
20 <212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 36 Met Ser Leu Leu Thr Glu Val Glu Thr Pro
Ile Arg Asn Glu Thr Pro 1 5 10 15 Ile Arg Asn Glu 20 <210>
SEQ ID NO 37 <211> LENGTH: 15 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 37 Met
Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp 1 5 10 15
<210> SEQ ID NO 38 <211> LENGTH: 16 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 38
Met Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly 1 5
10 15 <210> SEQ ID NO 39 <211> LENGTH: 24 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 39 Met Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn
Glu Trp Gly 1 5 10 15 Cys Arg Cys Asn Asp Ser Ser Asp 20
<210> SEQ ID NO 40 <211> LENGTH: 8 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 40
Ser Leu Leu Thr Glu Val Glu Thr 1 5 <210> SEQ ID NO 41
<211> LENGTH: 13 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 41 Ser Leu Leu Thr Glu Val
Glu Thr Pro Ile Arg Asn Glu 1 5 10 <210> SEQ ID NO 42
<211> LENGTH: 14 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 42 Ser Leu Leu Thr Glu Val
Glu Thr Pro Ile Arg Asn Glu Trp 1 5 10 <210> SEQ ID NO 43
<211> LENGTH: 15 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 43 Ser Leu Leu Thr Glu Val
Glu Thr Pro Ile Arg Asn Glu Trp Gly 1 5 10 15 <210> SEQ ID NO
44 <211> LENGTH: 21 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 44 Ser Leu Leu Thr
Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly Thr 1 5 10 15 Pro Ile
Arg Asn Glu 20 <210> SEQ ID NO 45 <211> LENGTH: 27
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 45 Ser Leu Leu Thr Glu Val Glu Thr Pro Ile
Arg Asn Glu Trp Gly Thr 1 5 10 15 Pro Ile Arg Asn Glu Thr Pro Ile
Arg Asn Glu 20 25 <210> SEQ ID NO 46 <211> LENGTH: 33
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 46 Ser Leu Leu Thr Glu Val Glu Thr Pro Ile
Arg Asn Glu Trp Gly Thr 1 5 10 15 Pro Ile Arg Asn Glu Thr Pro Ile
Arg Asn Glu Thr Pro Ile Arg Asn 20 25 30 Glu <210> SEQ ID NO
47 <211> LENGTH: 39 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 47 Ser Leu Leu Thr
Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly Leu 1 5 10 15 Leu Thr
Glu Val Glu Thr Pro Ile Arg Gln Tyr Ile Lys Ala Asn Ser 20 25 30
Lys Phe Ile Gly Ile Thr Glu 35 <210> SEQ ID NO 48 <211>
LENGTH: 10 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 48 Thr Glu Val Glu Thr Pro Ile Arg Asn
Glu 1 5 10 <210> SEQ ID NO 49 <211> LENGTH: 6
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 49 Thr Pro Ile Arg Asn Glu 1 5 <210>
SEQ ID NO 50 <211> LENGTH: 8 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 50 Val
Glu Thr Pro Ile Arg Asn Glu 1 5 <210> SEQ ID NO 51
<211> LENGTH: 52 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 51 Val Thr Arg Glu Pro Tyr
Val Ser Cys Asp Pro Lys Ser Cys Ile Asn 1 5 10 15 Arg Cys Phe Tyr
Val Glu Leu Ile Arg Gly Arg Val Thr Arg Glu Pro 20 25 30 Tyr Val
Ser Cys Asp Pro Trp Tyr Ile Lys Ala Asn Ser Lys Phe Ile 35 40 45
Gly Ile Thr Glu 50 <210> SEQ ID NO 52 <211> LENGTH: 6
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 52 Trp Gly Ile His His Pro 1 5 <210>
SEQ ID NO 53 <211> LENGTH: 6 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 53 Trp
Gly Val His His Pro 1 5 <210> SEQ ID NO 54 <211>
LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 54 Trp Gly Val Ile His His Pro 1 5
<210> SEQ ID NO 55 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 55
Trp Gly Ile Val His His Pro 1 5 <210> SEQ ID NO 56
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 56 Tyr Ile Trp Gly Ile His
His Pro 1 5 <210> SEQ ID NO 57 <211> LENGTH: 8
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 57 Tyr Ile Trp Gly Val His His Pro 1 5
<210> SEQ ID NO 58 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 58
Tyr Ile Trp Gly Val Ile His His Pro 1 5 <210> SEQ ID NO 59
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 59 Tyr Ile Trp Gly Ile Val
His His Pro 1 5 <210> SEQ ID NO 60 <211> LENGTH: 14
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 60 Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile
Gly Ile Thr Glu 1 5 10 <210> SEQ ID NO 61 <211> LENGTH:
15 <212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 61 Pro Ile Arg Asn Glu Trp Gly Cys Arg Cys
Asn Asp Ser Ser Asp 1 5 10 15 <210> SEQ ID NO 62 <400>
SEQUENCE: 62 000 <210> SEQ ID NO 63 <400> SEQUENCE: 63
000 <210> SEQ ID NO 64 <400> SEQUENCE: 64 000
<210> SEQ ID NO 65 <211> LENGTH: 372 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 65 Ser Glu Tyr Ala Tyr
Gly Ser Phe Val Arg Thr Val Ser Leu Pro Val 1 5 10 15 Gly Ala Asp
Glu Gly Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His 20 25 30 His
Pro His Tyr Glu Glu Cys Ser Cys Tyr Gly Leu Pro Val Glu Tyr 35 40
45 Leu Gln Val Pro Ser Pro Ser Met Gly Arg Asp Ile Lys Val Gln Phe
50 55 60 Gln Ser Gly Gly Ala Asn Ser Pro Ala Leu Tyr Leu Leu Asp
Gly Leu 65 70 75 80 Arg Ala Gln Asp Asp Phe Ser Gly Trp Asp Ile Asn
Thr Pro Ala Phe 85 90 95 Glu Trp Tyr Asp Gln Ser Gly Leu Ser Val
Val Met Pro Val Gly Gly 100 105 110 Gln Ser Ser Phe Tyr Ser Asp Trp
Tyr Gln Pro Ala Cys Arg Lys Ala 115 120 125 Gly Cys Gln Thr Tyr Lys
Trp Glu Thr Phe Leu Thr Ser Glu Leu Pro 130 135 140 Gly Trp Leu Gln
Ala Asn Arg His Val Gln Pro Thr Gly Ser Ala Val 145 150 155 160 Val
Gly Leu Ser Met Ala Ala Ser Ser Ala Leu Thr Leu Ala Ile Tyr 165 170
175 His Pro Gln Gln Phe Val Tyr Ala Gly Ala Met Ser Gly Leu Leu Asp
180 185 190 Pro Ser Gln Ala Met Gly Pro Thr Leu Ile Gly Leu Ala Met
Gly Asp 195 200 205 Ala Gly Gly Tyr Lys Ala Ser Asp Met Trp Gly Pro
Lys Glu Asp Pro 210 215 220 Ala Trp Gln Arg Asn Asp Pro Leu Leu Asn
Val Gly Lys Leu Ile Ala 225 230 235 240 Asn Asn Thr Arg Val Trp Val
Tyr Cys Gly Asn Gly Lys Pro Ser Asp 245 250 255 Leu Gly Gly Asn Asn
Leu Pro Ala Lys Phe Leu Glu Gly Phe Val Arg 260 265 270 Thr Ser Asn
Ile Lys Phe Gln Asp Ala Tyr Asn Ala Gly Gly His Asn 275 280 285 Gly
Val Phe Asp Phe Pro Asp Ser Gly Thr His Ser Trp Glu Tyr Trp 290 295
300 Gly Ala Gln Leu Asn Ala Met Lys Pro Asp Leu Gln Arg His Trp Val
305 310 315 320 Pro Arg Pro Thr Pro Gly Pro Pro Gln Gly Ala Phe Asp
Phe Pro Asp 325 330 335 Ser Gly Thr His Ser Trp Glu Tyr Trp Gly Ala
Gln Leu Asn Ala Met 340 345 350 Lys Pro Asp Leu Gln Arg His Trp Val
Pro Arg Pro Thr Pro Gly Pro 355 360 365 Pro Gln Gly Ala 370
<210> SEQ ID NO 66 <211> LENGTH: 22 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 66 Gly Asn Leu Phe Ile Ala
Pro Trp Gly Val Ile His His Pro His Tyr 1 5 10 15 Glu Glu Cys Ser
Cys Tyr 20 <210> SEQ ID NO 67 <211> LENGTH: 22
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 67 Trp
Gly Val Ile His His Pro Gly Asn Leu Phe Ile Ala Pro His Tyr 1 5 10
15 Glu Glu Cys Ser Cys Tyr 20 <210> SEQ ID NO 68 <211>
LENGTH: 294 <212> TYPE: PRT <213> ORGANISM:
Mycobacterium tuberculosis <400> SEQUENCE: 68 Ser Arg Pro Gly
Leu Pro Val Glu Tyr Leu Gln Val Pro Ser Pro Ser 1 5 10 15 Met Gly
Arg Asp Ile Lys Val Gln Phe Gln Ser Gly Gly Ala Asn Ser 20 25 30
Pro Ala Leu Tyr Leu Leu Asp Gly Leu Arg Ala Gln Asp Asp Phe Ser 35
40 45 Gly Trp Asp Ile Asn Thr Pro Ala Phe Glu Trp Tyr Asp Gln Ser
Gly 50 55 60 Leu Ser Val Val Met Pro Val Gly Gly Gln Ser Ser Phe
Tyr Ser Asp 65 70 75 80 Trp Tyr Gln Pro Ala Cys Gly Lys Ala Gly Cys
Gln Thr Tyr Lys Trp 85 90 95 Glu Thr Phe Leu Thr Ser Glu Leu Pro
Gly Trp Leu Gln Ala Asn Arg 100 105 110 His Val Lys Pro Thr Gly Ser
Ala Val Val Gly Leu Ser Met Ala Ala 115 120 125 Ser Ser Ala Leu Thr
Leu Ala Ile Tyr His Pro Gln Gln Phe Val Tyr 130 135 140 Ala Gly Ala
Met Ser Gly Leu Leu Asp Pro Ser Gln Ala Met Gly Pro 145 150 155 160
Thr Leu Ile Gly Leu Ala Met Gly Asp Ala Gly Gly Tyr Lys Ala Ser 165
170 175 Asp Met Trp Gly Pro Lys Glu Asp Pro Ala Trp Gln Arg Asn Asp
Pro 180 185 190 Leu Leu Asn Val Gly Lys Leu Ile Ala Asn Asn Thr Arg
Val Trp Val 195 200 205 Tyr Cys Gly Asn Gly Lys Pro Ser Asp Leu Gly
Gly Asn Asn Leu Pro 210 215 220 Ala Lys Phe Leu Glu Gly Phe Val Arg
Thr Ser Asn Ile Lys Phe Gln 225 230 235 240 Asp Ala Tyr Asn Ala Gly
Gly Gly His Asn Gly Val Phe Asp Phe Pro 245 250 255 Asp Ser Gly Thr
His Ser Trp Glu Tyr Trp Gly Ala Gln Leu Asn Ala 260 265 270 Met Lys
Pro Asp Leu Gln Arg Ala Leu Gly Ala Thr Pro Asn Thr Gly 275 280 285
Pro Ala Pro Gln Gly Ala 290 <210> SEQ ID NO 69 <211>
LENGTH: 882 <212> TYPE: DNA <213> ORGANISM:
Mycobacterium tuberculosis <400> SEQUENCE: 69 tcccggccgg
gcttgccggt ggagtacctg caggtgccgt cgccgtcgat gggccgtgac 60
atcaaggtcc aattccaaag tggtggtgcc aactcgcccg ccctgtacct gctcgacggc
120 ctgcgcgcgc aggacgactt cagcggctgg gacatcaaca ccccggcgtt
cgagtggtac 180 gaccagtcgg gcctgtcggt ggtcatgccg gtgggtggcc
agtcaagctt ctactccgac 240 tggtaccagc ccgcctgcgg caaggccggt
tgccagactt acaagtggga gaccttcctg 300 accagcgagc tgccggggtg
gctgcaggcc aacaggcacg tcaagcccac cggaagcgcc 360 gtcgtcggtc
tttcgatggc tgcttcttcg gcgctgacgc tggcgatcta tcacccccag 420
cagttcgtct acgcgggagc gatgtcgggc ctgttggacc cctcccaggc gatgggtccc
480 accctgatcg gcctggcgat gggtgacgct ggcggctaca aggcctccga
catgtggggc 540 ccgaaggagg acccggcgtg gcagcgcaac gacccgctgt
tgaacgtcgg gaagctgatc 600 gccaacaaca cccgcgtctg ggtgtactgc
ggcaacggca agccgtcgga tctgggtggc 660 aacaacctgc cggccaagtt
cctcgagggc ttcgtgcgga ccagcaacat caagttccaa 720 gacgcctaca
acgccggtgg cggccacaac ggcgtgttcg acttcccgga cagcggtacg 780
cacagctggg agtactgggg cgcgcagctc aacgctatga agcccgacct gcaacgggca
840 ctgggtgcca cgcccaacac cgggcccgcg ccccagggcg cc 882 <210>
SEQ ID NO 70 <211> LENGTH: 336 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 70 Ser Glu Phe Ala Tyr
Gly Ser Phe Val Arg Thr Val Ser Leu Pro Val 1 5 10 15 Gly Ala Asp
Glu Gly Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His 20 25 30 His
Pro His Tyr Glu Glu Cys Ser Cys Tyr Ser Arg Pro Gly Leu Pro 35 40
45 Val Glu Tyr Leu Gln Val Pro Ser Pro Ser Met Gly Arg Asp Ile Lys
50 55 60 Val Gln Phe Gln Ser Gly Gly Ala Asn Ser Pro Ala Leu Tyr
Leu Leu 65 70 75 80 Asp Gly Leu Arg Ala Gln Asp Asp Phe Ser Gly Trp
Asp Ile Asn Thr 85 90 95 Pro Ala Phe Glu Trp Tyr Asp Gln Ser Gly
Leu Ser Val Val Met Pro 100 105 110 Val Gly Gly Gln Ser Ser Phe Tyr
Ser Asp Trp Tyr Gln Pro Ala Cys 115 120 125 Gly Lys Ala Gly Cys Gln
Thr Tyr Lys Trp Glu Thr Phe Leu Thr Ser 130 135 140 Glu Leu Pro Gly
Trp Leu Gln Ala Asn Arg His Val Lys Pro Thr Gly 145 150 155 160 Ser
Ala Val Val Gly Leu Ser Met Ala Ala Ser Ser Ala Leu Thr Leu 165 170
175 Ala Ile Tyr His Pro Gln Gln Phe Val Tyr Ala Gly Ala Met Ser Gly
180 185 190 Leu Leu Asp Pro Ser Gln Ala Met Gly Pro Thr Leu Ile Gly
Leu Ala 195 200 205 Met Gly Asp Ala Gly Gly Tyr Lys Ala Ser Asp Met
Trp Gly Pro Lys 210 215 220 Glu Asp Pro Ala Trp Gln Arg Asn Asp Pro
Leu Leu Asn Val Gly Lys 225 230 235 240 Leu Ile Ala Asn Asn Thr Arg
Val Trp Val Tyr Cys Gly Asn Gly Lys 245 250 255 Pro Ser Asp Leu Gly
Gly Asn Asn Leu Pro Ala Lys Phe Leu Glu Gly 260 265 270 Phe Val Arg
Thr Ser Asn Ile Lys Phe Gln Asp Ala Tyr Asn Ala Gly 275 280 285 Gly
Gly His Asn Gly Val Phe Asp Phe Pro Asp Ser Gly Thr His Ser 290 295
300 Trp Glu Tyr Trp Gly Ala Gln Leu Asn Ala Met Lys Pro Asp Leu Gln
305 310 315 320 Arg Ala Leu Gly Ala Thr Pro Asn Thr Gly Pro Ala Pro
Gln Gly Ala 325 330 335 <210> SEQ ID NO 71 <211>
LENGTH: 1038 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic polynucleotide
<400> SEQUENCE: 71 tttgggccca ttatgtcgga attcgcgtac
ggttccttcg ttcgcacggt gtcgctgccg 60 gtaggtgctg acgaggggaa
tctattcatt gctccttggg gggttattca ccacccgcat 120 tatgaggaat
gttcctgtta ctcccggccg ggcttgccgg tggagtacct gcaggtgccg 180
tcgccgtcga tgggccgtga catcaaggtc caattccaaa gtggtggtgc caactcgccc
240 gccctgtacc tgctcgacgg cctgcgcgcg caggacgact tcagcggctg
ggacatcaac 300 accccggcgt tcgagtggta cgaccagtcg ggcctgtcgg
tggtcatgcc ggtgggtggc 360 cagtcaagct tctactccga ctggtaccag
cccgcctgcg gcaaggccgg ttgccagact 420 tacaagtggg agaccttcct
gaccagcgag ctgccggggt ggctgcaggc caacaggcac 480 gtcaagccca
ccggaagcgc cgtcgtcggt ctttcgatgg ctgcttcttc ggcgctgacg 540
ctggcgatct atcaccccca gcagttcgtc tacgcgggag cgatgtcggg cctgttggac
600 ccctcccagg cgatgggtcc caccctgatc ggcctggcga tgggtgacgc
tggcggctac 660 aaggcctccg acatgtgggg cccgaaggag gacccggcgt
ggcagcgcaa cgacccgctg 720 ttgaacgtcg ggaagctgat cgccaacaac
acccgcgtct gggtgtactg cggcaacggc 780 aagccgtcgg atctgggtgg
caacaacctg ccggccaagt tcctcgaggg cttcgtgcgg 840 accagcaaca
tcaagttcca agacgcctac aacgccggtg gcggccacaa cggcgtgttc 900
gacttcccgg acagcggtac gcacagctgg gagtactggg gcgcgcagct caacgctatg
960 aagcccgacc tgcaacgggc actgggtgcc acgcccaaca ccgggcccgc
gccccagggc 1020 gcctagtttc ttaagttt 1038 <210> SEQ ID NO 72
<211> LENGTH: 21 <212> TYPE: DNA <213> ORGANISM:
Influenza virus <400> SEQUENCE: 72 gggaatctat tcattgctcc t 21
<210> SEQ ID NO 73 <211> LENGTH: 21 <212> TYPE:
DNA <213> ORGANISM: Influenza virus <400> SEQUENCE: 73
tggggggtta ttcaccaccc g 21 <210> SEQ ID NO 74 <211>
LENGTH: 24 <212> TYPE: DNA <213> ORGANISM: Influenza
virus <400> SEQUENCE: 74 cattatgagg aatgttcctg ttac 24
<210> SEQ ID NO 75 <211> LENGTH: 20 <212> TYPE:
PRT <213> ORGANISM: Mycobacterium tuberculosis <400>
SEQUENCE: 75 Ser Glu Phe Ala Tyr Gly Ser Phe Val Arg Thr Val Ser
Leu Pro Val 1 5 10 15 Gly Ala Asp Glu 20 <210> SEQ ID NO 76
<211> LENGTH: 60 <212> TYPE: DNA <213> ORGANISM:
Mycobacterium tuberculosis <400> SEQUENCE: 76 tcggaattcg
cgtacggttc cttcgttcgc acggtgtcgc tgccggtagg tgctgacgag 60
<210> SEQ ID NO 77 <211> LENGTH: 22 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 77 Gly Asn Leu Phe Ile Ala
Pro Trp Gly Val Ile His His Pro His Tyr 1 5 10 15 Glu Glu Cys Ser
Cys Tyr 20 <210> SEQ ID NO 78 <211> LENGTH: 66
<212> TYPE: DNA <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic oligonucleotide <400>
SEQUENCE: 78 gggaatctat tcattgctcc ttggggggtt attcaccacc cgcattatga
ggaatgttcc 60 tgttac 66 <210> SEQ ID NO 79 <211>
LENGTH: 1008 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic polynucleotide
<400> SEQUENCE: 79 tcggaattcg cgtacggttc cttcgttcgc
acggtgtcgc tgccggtagg tgctgacgag 60 gggaatctat tcattgctcc
ttggggggtt attcaccacc cgcattatga ggaatgttcc 120 tgttactccc
ggccgggctt gccggtggag tacctgcagg tgccgtcgcc gtcgatgggc 180
cgtgacatca aggtccaatt ccaaagtggt ggtgccaact cgcccgccct gtacctgctc
240 gacggcctgc gcgcgcagga cgacttcagc ggctgggaca tcaacacccc
ggcgttcgag 300 tggtacgacc agtcgggcct gtcggtggtc atgccggtgg
gtggccagtc aagcttctac 360 tccgactggt accagcccgc ctgcggcaag
gccggttgcc agacttacaa gtgggagacc 420 ttcctgacca gcgagctgcc
ggggtggctg caggccaaca ggcacgtcaa gcccaccgga 480 agcgccgtcg
tcggtctttc gatggctgct tcttcggcgc tgacgctggc gatctatcac 540
ccccagcagt tcgtctacgc gggagcgatg tcgggcctgt tggacccctc ccaggcgatg
600 ggtcccaccc tgatcggcct ggcgatgggt gacgctggcg gctacaaggc
ctccgacatg 660 tggggcccga aggaggaccc ggcgtggcag cgcaacgacc
cgctgttgaa cgtcgggaag 720 ctgatcgcca acaacacccg cgtctgggtg
tactgcggca acggcaagcc gtcggatctg 780 ggtggcaaca acctgccggc
caagttcctc gagggcttcg tgcggaccag caacatcaag 840 ttccaagacg
cctacaacgc cggtggcggc cacaacggcg tgttcgactt cccggacagc 900
ggtacgcaca gctgggagta ctggggcgcg cagctcaacg ctatgaagcc cgacctgcaa
960 cgggcactgg gtgccacgcc caacaccggg cccgcgcccc agggcgcc 1008
<210> SEQ ID NO 80 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 80 Cys Ala Gly Ala Gly Asn
Phe Ile Ala Pro 1 5 10 <210> SEQ ID NO 81 <211> LENGTH:
10 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 81 Cys
Ala Gly Ala Gly Asn Leu Ile Ala Pro 1 5 10 <210> SEQ ID NO 82
<211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 82 Cys Ala Gly Ala Gly Asn Leu Phe Ile Ala
Pro 1 5 10 <210> SEQ ID NO 83 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 83 Cys
Ala Gly Ala Trp Gly Val His His Pro 1 5 10 <210> SEQ ID NO 84
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 84 Cys Ala Gly Ala Trp Gly Ile His His Pro 1
5 10 <210> SEQ ID NO 85 <211> LENGTH: 11 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 85 Cys Ala Gly
Ala Trp Gly Val Ile His His Pro 1 5 10 <210> SEQ ID NO 86
<211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 86 Cys Ala Gly Ala Trp Gly Ile Val His His
Pro 1 5 10 <210> SEQ ID NO 87 <211> LENGTH: 13
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 87 Gly
Asn Leu Ile Ala Pro Trp Gly Val Ile His His Pro 1 5 10 <210>
SEQ ID NO 88 <211> LENGTH: 17 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 88 Cys Ala Gly Ala Gly Asn
Leu Ile Ala Pro Trp Gly Val Ile His His 1 5 10 15 Pro <210>
SEQ ID NO 89 <211> LENGTH: 14 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 89 Gly Asn Leu Phe Ile Ala
Pro Trp Gly Val Ile His His Pro 1 5 10 <210> SEQ ID NO 90
<211> LENGTH: 18 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 90 Cys Ala Gly Ala Gly Asn Leu Phe Ile Ala
Pro Trp Gly Val Ile His 1 5 10 15 His Pro <210> SEQ ID NO 91
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 91 His Tyr Glu Glu Cys Ser Cys Tyr 1 5
<210> SEQ ID NO 92 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 92 Cys Ala Gly Ala His Tyr
Glu Glu Cys Ser Cys Tyr 1 5 10 <210> SEQ ID NO 93 <211>
LENGTH: 22 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 93 Gly Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His His
Pro His Tyr 1 5 10 15 Glu Glu Cys Ser Cys Tyr 20 <210> SEQ ID
NO 94 <211> LENGTH: 26 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 94 Cys Ala Gly Ala Gly Asn Leu Phe
Ile Ala Pro Trp Gly Val Ile His 1 5 10 15 His Pro His Tyr Glu Glu
Cys Ser Cys Tyr 20 25 <210> SEQ ID NO 95 <211> LENGTH:
28 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 95 Gly
Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His His Pro Gly Asn 1 5 10
15 Leu Phe Ile Ala Pro Trp Gly Val Ile His His Pro 20 25
<210> SEQ ID NO 96 <211> LENGTH: 32 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 96 Cys Ala Gly Ala Gly
Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His 1 5 10 15 His Pro Gly
Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His His Pro 20 25 30
<210> SEQ ID NO 97 <211> LENGTH: 22 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 97 His Tyr Glu Glu Cys Ser
Cys Tyr Gly Asn Leu Phe Ile Ala Pro Trp 1 5 10 15 Gly Val Ile His
His Pro 20 <210> SEQ ID NO 98 <211> LENGTH: 22
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 98 Gly
Asn Leu Phe Ile Ala Pro His Tyr Glu Glu Cys Ser Cys Tyr Trp 1 5 10
15 Gly Val Ile His His Pro 20 <210> SEQ ID NO 99 <211>
LENGTH: 39 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic polypeptide
<400> SEQUENCE: 99 Ser Leu Leu Thr Glu Val Glu Thr Pro Ile
Arg Asn Glu Trp Gly Leu 1 5 10 15 Leu Thr Glu Val Glu Thr Pro Ile
Arg Gln Tyr Ile Lys Ala Asn Ser 20 25 30 Lys Phe Ile Gly Ile Thr
Glu 35 <210> SEQ ID NO 100 <211> LENGTH: 35 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic polypeptide <400> SEQUENCE: 100 Gly Asn
Leu Phe Ile Ala Pro Gly Asn Leu Phe Ile Ala Pro Gln Tyr 1 5 10 15
Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu Gly Asn Leu Phe 20
25 30 Ile Ala Pro 35 <210> SEQ ID NO 101 <211> LENGTH:
48 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
101 His Tyr Glu Glu Cys Ser Cys Tyr Asp Trp Ser Gly Tyr Ser Gly Ser
1 5 10 15 Phe Val Gln His Pro Glu Leu Thr Gly Leu His Tyr Glu Glu
Cys Ser 20 25 30 Cys Tyr Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile
Gly Ile Thr Glu 35 40 45 <210> SEQ ID NO 102 <211>
LENGTH: 52 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic polypeptide
<400> SEQUENCE: 102 Val Thr Arg Glu Pro Tyr Val Ser Cys Asp
Pro Lys Ser Cys Ile Asn 1 5 10 15 Arg Cys Phe Tyr Val Glu Leu Ile
Arg Gly Arg Val Thr Arg Glu Pro 20 25 30 Tyr Val Ser Cys Asp Pro
Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile 35 40 45 Gly Ile Thr Glu 50
<210> SEQ ID NO 103 <211> LENGTH: 18 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 103 Asp Trp Ser Gly Tyr Ser
Gly Ser Phe Val Gln His Pro Glu Leu Thr 1 5 10 15 Gly Leu
<210> SEQ ID NO 104 <211> LENGTH: 33 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 104 Ile Thr Gly Phe Ala
Pro Phe Ser Lys Asp Asn Ser Ile Arg Leu Ser 1 5 10 15 Ala Gly Gly
Asp Ile Trp Val Thr Arg Glu Pro Tyr Val Ser Cys Asp 20 25 30 Pro
<210> SEQ ID NO 105 <211> LENGTH: 16 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 105 Lys Ser Cys Ile Asn Arg
Cys Phe Tyr Val Glu Leu Ile Arg Gly Arg 1 5 10 15 <210> SEQ
ID NO 106 <211> LENGTH: 12 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 106 Gly Asn Leu Phe Ile Ala Pro Arg
Tyr Ala Phe Ala 1 5 10 <210> SEQ ID NO 107 <211>
LENGTH: 16 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 107 Cys Ala Gly Ala Gly Asn Leu Phe Ile Ala Pro Arg Tyr
Ala Phe Ala 1 5 10 15 <210> SEQ ID NO 108 <211> LENGTH:
11 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 108
Gly Asn Leu Val Val Pro Arg Tyr Ala Phe Ala 1 5 10 <210> SEQ
ID NO 109 <211> LENGTH: 15 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 109 Cys Ala Gly Ala Gly Asn Leu Val
Val Pro Arg Tyr Ala Phe Ala 1 5 10 15 <210> SEQ ID NO 110
<211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 110 Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe
Ala 1 5 10 <210> SEQ ID NO 111 <211> LENGTH: 15
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 111
Cys Ala Gly Ala Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala 1 5 10
15 <210> SEQ ID NO 112 <211> LENGTH: 6 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 112 Gly Asn Leu
Val Val Pro 1 5 <210> SEQ ID NO 113 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 113
Cys Ala Gly Ala Gly Asn Leu Val Val Pro 1 5 10 <210> SEQ ID
NO 114 <211> LENGTH: 15 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 114 Phe Val Ile Arg Glu Pro Phe Ile
Ser Cys Ser His Leu Glu Cys 1 5 10 15 <210> SEQ ID NO 115
<211> LENGTH: 19 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 115 Cys Ala Gly Ala Phe Val Ile Arg Glu Pro
Phe Ile Ser Cys Ser His 1 5 10 15 Leu Glu Cys <210> SEQ ID NO
116 <211> LENGTH: 4 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 116 Cys Ala Gly Ala 1
1 SEQUENCE LISTING <160> NUMBER OF SEQ ID NOS: 116
<210> SEQ ID NO 1 <211> LENGTH: 19 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 1
Asp Trp Ser Gly Tyr Ser Gly Ser Phe Val Gln His Pro Glu Leu Thr 1 5
10 15 Gly Leu Asp <210> SEQ ID NO 2 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 2 Glu Thr Pro Ile Arg Asn Glu 1 5 <210>
SEQ ID NO 3 <211> LENGTH: 15 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 3 Phe
Val Ile Arg Glu Pro Phe Ile Ser Cys Ser His Leu Glu Cys 1 5 10 15
<210> SEQ ID NO 4 <211> LENGTH: 6 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 4 Gly
Asn Phe Ile Ala Pro 1 5 <210> SEQ ID NO 5 <211> LENGTH:
6 <212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 5 Gly Asn Leu Ile Ala Pro 1 5 <210> SEQ
ID NO 6 <211> LENGTH: 7 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 6 Gly Asn Leu Phe
Ile Ala Pro 1 5 <210> SEQ ID NO 7 <211> LENGTH: 7
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 7 Gly Asn Leu Ile Phe Ala Pro 1 5 <210>
SEQ ID NO 8 <211> LENGTH: 8 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 8 His Tyr Glu Glu
Cys Ser Cys Tyr 1 5 <210> SEQ ID NO 9 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 9 Leu Leu Thr Glu Val Glu Thr Pro Ile Arg 1 5
10 <210> SEQ ID NO 10 <211> LENGTH: 11 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 10 Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn 1 5 10
<210> SEQ ID NO 11 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 11
Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu 1 5 10 <210>
SEQ ID NO 12 <211> LENGTH: 18 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 12 Asp
Trp Ser Gly Tyr Ser Gly Ser Phe Val Gln His Pro Glu Leu Thr 1 5 10
15 Gly Leu <210> SEQ ID NO 13 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 13 Glu Val Glu Thr Pro Ile Arg Asn Glu 1 5
<210> SEQ ID NO 14 <211> LENGTH: 38 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 14
Phe Leu Leu Pro Glu Asp Glu Thr Pro Ile Arg Asn Glu Trp Gly Leu 1 5
10 15 Leu Thr Asp Asp Glu Thr Pro Ile Arg Tyr Ile Lys Ala Asn Ser
Lys 20 25 30 Phe Ile Gly Ile Thr Glu 35 <210> SEQ ID NO 15
<211> LENGTH: 64 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 15 Gly Asn Leu Phe Ile Ala
Pro Gly Asn Leu Phe Ile Ala Pro His Tyr 1 5 10 15 Glu Glu Cys Ser
Cys Tyr His Tyr Glu Glu Cys Ser Cys Tyr Gln Tyr 20 25 30 Ile Lys
Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu His Tyr Glu Glu 35 40 45
Cys Ser Cys Tyr Thr Pro Ile Arg Asn Glu Thr Pro Ile Arg Asn Glu 50
55 60 <210> SEQ ID NO 16 <211> LENGTH: 35 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 16 Gly Asn Leu Phe Ile Ala Pro Gly Asn Leu Phe Ile Ala
Pro Gln Tyr 1 5 10 15 Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr
Glu Gly Asn Leu Phe 20 25 30 Ile Ala Pro 35 <210> SEQ ID NO
17 <211> LENGTH: 48 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 17 His Tyr Glu Glu
Cys Ser Cys Tyr Asp Trp Ser Gly Tyr Ser Gly Ser 1 5 10 15 Phe Val
Gln His Pro Glu Leu Thr Gly Leu His Tyr Glu Glu Cys Ser 20 25 30
Cys Tyr Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu 35
40 45 <210> SEQ ID NO 18 <211> LENGTH: 33 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 18 Ile Thr Gly Phe Ala Pro Phe Ser Lys Asp Asn Ser Ile
Arg Leu Ser 1 5 10 15 Ala Gly Gly Asp Ile Trp Val Thr Arg Glu Pro
Tyr Val Ser Cys Asp 20 25 30 Pro <210> SEQ ID NO 19
<211> LENGTH: 7 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 19 Ile Trp Gly Ile His His
Pro 1 5
<210> SEQ ID NO 20 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 20
Ile Trp Gly Val His His Pro 1 5 <210> SEQ ID NO 21
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 21 Ile Trp Gly Val Ile His
His Pro 1 5 <210> SEQ ID NO 22 <211> LENGTH: 8
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 22 Ile Trp Gly Ile Val His His Pro 1 5
<210> SEQ ID NO 23 <211> LENGTH: 16 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 23
Lys Ser Cys Ile Asn Arg Cys Phe Tyr Val Glu Leu Ile Arg Gly Arg 1 5
10 15 <210> SEQ ID NO 24 <211> LENGTH: 27 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 24 Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Ser
Leu Leu Thr 1 5 10 15 Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly
20 25 <210> SEQ ID NO 25 <211> LENGTH: 13 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 25 Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp 1
5 10 <210> SEQ ID NO 26 <211> LENGTH: 14 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 26 Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp
Gly 1 5 10 <210> SEQ ID NO 27 <211> LENGTH: 11
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 27 Leu Thr Glu Val Glu Thr Pro Ile Arg Asn
Glu 1 5 10 <210> SEQ ID NO 28 <211> LENGTH: 12
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 28 Leu Thr Glu Val Glu Thr Pro Ile Arg Asn
Glu Trp 1 5 10 <210> SEQ ID NO 29 <211> LENGTH: 13
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 29 Leu Thr Glu Val Glu Thr Pro Ile Arg Asn
Glu Trp Gly 1 5 10 <210> SEQ ID NO 30 <211> LENGTH: 9
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 30 Met Ser Leu Leu Thr Glu Val Glu Thr 1 5
<210> SEQ ID NO 31 <211> LENGTH: 10 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 31
Met Ser Leu Leu Thr Glu Val Glu Thr Pro 1 5 10 <210> SEQ ID
NO 32 <211> LENGTH: 11 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 32 Met Ser Leu Leu
Thr Glu Val Glu Thr Pro Ile 1 5 10 <210> SEQ ID NO 33
<211> LENGTH: 12 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 33 Met Ser Leu Leu Thr Glu
Val Glu Thr Pro Ile Arg 1 5 10 <210> SEQ ID NO 34 <211>
LENGTH: 13 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 34 Met Ser Leu Leu Thr Glu Val Glu Thr
Pro Ile Arg Asn 1 5 10 <210> SEQ ID NO 35 <211> LENGTH:
14 <212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 35 Met Ser Leu Leu Thr Glu Val Glu Thr Pro
Ile Arg Asn Glu 1 5 10 <210> SEQ ID NO 36 <211> LENGTH:
20 <212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 36 Met Ser Leu Leu Thr Glu Val Glu Thr Pro
Ile Arg Asn Glu Thr Pro 1 5 10 15 Ile Arg Asn Glu 20 <210>
SEQ ID NO 37 <211> LENGTH: 15 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 37 Met
Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp 1 5 10 15
<210> SEQ ID NO 38 <211> LENGTH: 16 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 38
Met Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly 1 5
10 15 <210> SEQ ID NO 39 <211> LENGTH: 24 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 39 Met Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn
Glu Trp Gly 1 5 10 15 Cys Arg Cys Asn Asp Ser Ser Asp 20
<210> SEQ ID NO 40 <211> LENGTH: 8 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 40
Ser Leu Leu Thr Glu Val Glu Thr 1 5 <210> SEQ ID NO 41
<211> LENGTH: 13 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 41 Ser Leu Leu Thr Glu Val
Glu Thr Pro Ile Arg Asn Glu 1 5 10
<210> SEQ ID NO 42 <211> LENGTH: 14 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 42
Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp 1 5 10
<210> SEQ ID NO 43 <211> LENGTH: 15 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 43
Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly 1 5 10
15 <210> SEQ ID NO 44 <211> LENGTH: 21 <212>
TYPE: PRT <213> ORGANISM: Influenza virus <400>
SEQUENCE: 44 Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu
Trp Gly Thr 1 5 10 15 Pro Ile Arg Asn Glu 20 <210> SEQ ID NO
45 <211> LENGTH: 27 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 45 Ser Leu Leu Thr
Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly Thr 1 5 10 15 Pro Ile
Arg Asn Glu Thr Pro Ile Arg Asn Glu 20 25 <210> SEQ ID NO 46
<211> LENGTH: 33 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 46 Ser Leu Leu Thr Glu Val
Glu Thr Pro Ile Arg Asn Glu Trp Gly Thr 1 5 10 15 Pro Ile Arg Asn
Glu Thr Pro Ile Arg Asn Glu Thr Pro Ile Arg Asn 20 25 30 Glu
<210> SEQ ID NO 47 <211> LENGTH: 39 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 47
Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly Leu 1 5
10 15 Leu Thr Glu Val Glu Thr Pro Ile Arg Gln Tyr Ile Lys Ala Asn
Ser 20 25 30 Lys Phe Ile Gly Ile Thr Glu 35 <210> SEQ ID NO
48 <211> LENGTH: 10 <212> TYPE: PRT <213>
ORGANISM: Influenza virus <400> SEQUENCE: 48 Thr Glu Val Glu
Thr Pro Ile Arg Asn Glu 1 5 10 <210> SEQ ID NO 49 <211>
LENGTH: 6 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 49 Thr Pro Ile Arg Asn Glu 1 5
<210> SEQ ID NO 50 <211> LENGTH: 8 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 50
Val Glu Thr Pro Ile Arg Asn Glu 1 5 <210> SEQ ID NO 51
<211> LENGTH: 52 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 51 Val Thr Arg Glu Pro Tyr
Val Ser Cys Asp Pro Lys Ser Cys Ile Asn 1 5 10 15 Arg Cys Phe Tyr
Val Glu Leu Ile Arg Gly Arg Val Thr Arg Glu Pro 20 25 30 Tyr Val
Ser Cys Asp Pro Trp Tyr Ile Lys Ala Asn Ser Lys Phe Ile 35 40 45
Gly Ile Thr Glu 50 <210> SEQ ID NO 52 <211> LENGTH: 6
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 52 Trp Gly Ile His His Pro 1 5 <210>
SEQ ID NO 53 <211> LENGTH: 6 <212> TYPE: PRT
<213> ORGANISM: Influenza virus <400> SEQUENCE: 53 Trp
Gly Val His His Pro 1 5 <210> SEQ ID NO 54 <211>
LENGTH: 7 <212> TYPE: PRT <213> ORGANISM: Influenza
virus <400> SEQUENCE: 54 Trp Gly Val Ile His His Pro 1 5
<210> SEQ ID NO 55 <211> LENGTH: 7 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 55
Trp Gly Ile Val His His Pro 1 5 <210> SEQ ID NO 56
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 56 Tyr Ile Trp Gly Ile His
His Pro 1 5 <210> SEQ ID NO 57 <211> LENGTH: 8
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 57 Tyr Ile Trp Gly Val His His Pro 1 5
<210> SEQ ID NO 58 <211> LENGTH: 9 <212> TYPE:
PRT <213> ORGANISM: Influenza virus <400> SEQUENCE: 58
Tyr Ile Trp Gly Val Ile His His Pro 1 5 <210> SEQ ID NO 59
<211> LENGTH: 9 <212> TYPE: PRT <213> ORGANISM:
Influenza virus <400> SEQUENCE: 59 Tyr Ile Trp Gly Ile Val
His His Pro 1 5 <210> SEQ ID NO 60 <211> LENGTH: 14
<212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 60 Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile
Gly Ile Thr Glu 1 5 10 <210> SEQ ID NO 61 <211> LENGTH:
15 <212> TYPE: PRT <213> ORGANISM: Influenza virus
<400> SEQUENCE: 61 Pro Ile Arg Asn Glu Trp Gly Cys Arg Cys
Asn Asp Ser Ser Asp 1 5 10 15 <210> SEQ ID NO 62 <400>
SEQUENCE: 62
000 <210> SEQ ID NO 63 <400> SEQUENCE: 63 000
<210> SEQ ID NO 64 <400> SEQUENCE: 64 000 <210>
SEQ ID NO 65 <211> LENGTH: 372 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 65 Ser Glu Tyr Ala Tyr
Gly Ser Phe Val Arg Thr Val Ser Leu Pro Val 1 5 10 15 Gly Ala Asp
Glu Gly Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His 20 25 30 His
Pro His Tyr Glu Glu Cys Ser Cys Tyr Gly Leu Pro Val Glu Tyr 35 40
45 Leu Gln Val Pro Ser Pro Ser Met Gly Arg Asp Ile Lys Val Gln Phe
50 55 60 Gln Ser Gly Gly Ala Asn Ser Pro Ala Leu Tyr Leu Leu Asp
Gly Leu 65 70 75 80 Arg Ala Gln Asp Asp Phe Ser Gly Trp Asp Ile Asn
Thr Pro Ala Phe 85 90 95 Glu Trp Tyr Asp Gln Ser Gly Leu Ser Val
Val Met Pro Val Gly Gly 100 105 110 Gln Ser Ser Phe Tyr Ser Asp Trp
Tyr Gln Pro Ala Cys Arg Lys Ala 115 120 125 Gly Cys Gln Thr Tyr Lys
Trp Glu Thr Phe Leu Thr Ser Glu Leu Pro 130 135 140 Gly Trp Leu Gln
Ala Asn Arg His Val Gln Pro Thr Gly Ser Ala Val 145 150 155 160 Val
Gly Leu Ser Met Ala Ala Ser Ser Ala Leu Thr Leu Ala Ile Tyr 165 170
175 His Pro Gln Gln Phe Val Tyr Ala Gly Ala Met Ser Gly Leu Leu Asp
180 185 190 Pro Ser Gln Ala Met Gly Pro Thr Leu Ile Gly Leu Ala Met
Gly Asp 195 200 205 Ala Gly Gly Tyr Lys Ala Ser Asp Met Trp Gly Pro
Lys Glu Asp Pro 210 215 220 Ala Trp Gln Arg Asn Asp Pro Leu Leu Asn
Val Gly Lys Leu Ile Ala 225 230 235 240 Asn Asn Thr Arg Val Trp Val
Tyr Cys Gly Asn Gly Lys Pro Ser Asp 245 250 255 Leu Gly Gly Asn Asn
Leu Pro Ala Lys Phe Leu Glu Gly Phe Val Arg 260 265 270 Thr Ser Asn
Ile Lys Phe Gln Asp Ala Tyr Asn Ala Gly Gly His Asn 275 280 285 Gly
Val Phe Asp Phe Pro Asp Ser Gly Thr His Ser Trp Glu Tyr Trp 290 295
300 Gly Ala Gln Leu Asn Ala Met Lys Pro Asp Leu Gln Arg His Trp Val
305 310 315 320 Pro Arg Pro Thr Pro Gly Pro Pro Gln Gly Ala Phe Asp
Phe Pro Asp 325 330 335 Ser Gly Thr His Ser Trp Glu Tyr Trp Gly Ala
Gln Leu Asn Ala Met 340 345 350 Lys Pro Asp Leu Gln Arg His Trp Val
Pro Arg Pro Thr Pro Gly Pro 355 360 365 Pro Gln Gly Ala 370
<210> SEQ ID NO 66 <211> LENGTH: 22 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 66 Gly Asn Leu Phe Ile Ala
Pro Trp Gly Val Ile His His Pro His Tyr 1 5 10 15 Glu Glu Cys Ser
Cys Tyr 20 <210> SEQ ID NO 67 <211> LENGTH: 22
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 67 Trp
Gly Val Ile His His Pro Gly Asn Leu Phe Ile Ala Pro His Tyr 1 5 10
15 Glu Glu Cys Ser Cys Tyr 20 <210> SEQ ID NO 68 <211>
LENGTH: 294 <212> TYPE: PRT <213> ORGANISM:
Mycobacterium tuberculosis <400> SEQUENCE: 68 Ser Arg Pro Gly
Leu Pro Val Glu Tyr Leu Gln Val Pro Ser Pro Ser 1 5 10 15 Met Gly
Arg Asp Ile Lys Val Gln Phe Gln Ser Gly Gly Ala Asn Ser 20 25 30
Pro Ala Leu Tyr Leu Leu Asp Gly Leu Arg Ala Gln Asp Asp Phe Ser 35
40 45 Gly Trp Asp Ile Asn Thr Pro Ala Phe Glu Trp Tyr Asp Gln Ser
Gly 50 55 60 Leu Ser Val Val Met Pro Val Gly Gly Gln Ser Ser Phe
Tyr Ser Asp 65 70 75 80 Trp Tyr Gln Pro Ala Cys Gly Lys Ala Gly Cys
Gln Thr Tyr Lys Trp 85 90 95 Glu Thr Phe Leu Thr Ser Glu Leu Pro
Gly Trp Leu Gln Ala Asn Arg 100 105 110 His Val Lys Pro Thr Gly Ser
Ala Val Val Gly Leu Ser Met Ala Ala 115 120 125 Ser Ser Ala Leu Thr
Leu Ala Ile Tyr His Pro Gln Gln Phe Val Tyr 130 135 140 Ala Gly Ala
Met Ser Gly Leu Leu Asp Pro Ser Gln Ala Met Gly Pro 145 150 155 160
Thr Leu Ile Gly Leu Ala Met Gly Asp Ala Gly Gly Tyr Lys Ala Ser 165
170 175 Asp Met Trp Gly Pro Lys Glu Asp Pro Ala Trp Gln Arg Asn Asp
Pro 180 185 190 Leu Leu Asn Val Gly Lys Leu Ile Ala Asn Asn Thr Arg
Val Trp Val 195 200 205 Tyr Cys Gly Asn Gly Lys Pro Ser Asp Leu Gly
Gly Asn Asn Leu Pro 210 215 220 Ala Lys Phe Leu Glu Gly Phe Val Arg
Thr Ser Asn Ile Lys Phe Gln 225 230 235 240 Asp Ala Tyr Asn Ala Gly
Gly Gly His Asn Gly Val Phe Asp Phe Pro 245 250 255 Asp Ser Gly Thr
His Ser Trp Glu Tyr Trp Gly Ala Gln Leu Asn Ala 260 265 270 Met Lys
Pro Asp Leu Gln Arg Ala Leu Gly Ala Thr Pro Asn Thr Gly 275 280 285
Pro Ala Pro Gln Gly Ala 290 <210> SEQ ID NO 69 <211>
LENGTH: 882 <212> TYPE: DNA <213> ORGANISM:
Mycobacterium tuberculosis <400> SEQUENCE: 69 tcccggccgg
gcttgccggt ggagtacctg caggtgccgt cgccgtcgat gggccgtgac 60
atcaaggtcc aattccaaag tggtggtgcc aactcgcccg ccctgtacct gctcgacggc
120 ctgcgcgcgc aggacgactt cagcggctgg gacatcaaca ccccggcgtt
cgagtggtac 180 gaccagtcgg gcctgtcggt ggtcatgccg gtgggtggcc
agtcaagctt ctactccgac 240 tggtaccagc ccgcctgcgg caaggccggt
tgccagactt acaagtggga gaccttcctg 300 accagcgagc tgccggggtg
gctgcaggcc aacaggcacg tcaagcccac cggaagcgcc 360 gtcgtcggtc
tttcgatggc tgcttcttcg gcgctgacgc tggcgatcta tcacccccag 420
cagttcgtct acgcgggagc gatgtcgggc ctgttggacc cctcccaggc gatgggtccc
480 accctgatcg gcctggcgat gggtgacgct ggcggctaca aggcctccga
catgtggggc 540 ccgaaggagg acccggcgtg gcagcgcaac gacccgctgt
tgaacgtcgg gaagctgatc 600 gccaacaaca cccgcgtctg ggtgtactgc
ggcaacggca agccgtcgga tctgggtggc 660 aacaacctgc cggccaagtt
cctcgagggc ttcgtgcgga ccagcaacat caagttccaa 720 gacgcctaca
acgccggtgg cggccacaac ggcgtgttcg acttcccgga cagcggtacg 780
cacagctggg agtactgggg cgcgcagctc aacgctatga agcccgacct gcaacgggca
840 ctgggtgcca cgcccaacac cgggcccgcg ccccagggcg cc 882 <210>
SEQ ID NO 70 <211> LENGTH: 336 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 70
Ser Glu Phe Ala Tyr Gly Ser Phe Val Arg Thr Val Ser Leu Pro Val 1 5
10 15 Gly Ala Asp Glu Gly Asn Leu Phe Ile Ala Pro Trp Gly Val Ile
His 20 25 30 His Pro His Tyr Glu Glu Cys Ser Cys Tyr Ser Arg Pro
Gly Leu Pro 35 40 45 Val Glu Tyr Leu Gln Val Pro Ser Pro Ser Met
Gly Arg Asp Ile Lys 50 55 60 Val Gln Phe Gln Ser Gly Gly Ala Asn
Ser Pro Ala Leu Tyr Leu Leu 65 70 75 80 Asp Gly Leu Arg Ala Gln Asp
Asp Phe Ser Gly Trp Asp Ile Asn Thr 85 90 95 Pro Ala Phe Glu Trp
Tyr Asp Gln Ser Gly Leu Ser Val Val Met Pro 100 105 110 Val Gly Gly
Gln Ser Ser Phe Tyr Ser Asp Trp Tyr Gln Pro Ala Cys 115 120 125 Gly
Lys Ala Gly Cys Gln Thr Tyr Lys Trp Glu Thr Phe Leu Thr Ser 130 135
140 Glu Leu Pro Gly Trp Leu Gln Ala Asn Arg His Val Lys Pro Thr Gly
145 150 155 160 Ser Ala Val Val Gly Leu Ser Met Ala Ala Ser Ser Ala
Leu Thr Leu 165 170 175 Ala Ile Tyr His Pro Gln Gln Phe Val Tyr Ala
Gly Ala Met Ser Gly 180 185 190 Leu Leu Asp Pro Ser Gln Ala Met Gly
Pro Thr Leu Ile Gly Leu Ala 195 200 205 Met Gly Asp Ala Gly Gly Tyr
Lys Ala Ser Asp Met Trp Gly Pro Lys 210 215 220 Glu Asp Pro Ala Trp
Gln Arg Asn Asp Pro Leu Leu Asn Val Gly Lys 225 230 235 240 Leu Ile
Ala Asn Asn Thr Arg Val Trp Val Tyr Cys Gly Asn Gly Lys 245 250 255
Pro Ser Asp Leu Gly Gly Asn Asn Leu Pro Ala Lys Phe Leu Glu Gly 260
265 270 Phe Val Arg Thr Ser Asn Ile Lys Phe Gln Asp Ala Tyr Asn Ala
Gly 275 280 285 Gly Gly His Asn Gly Val Phe Asp Phe Pro Asp Ser Gly
Thr His Ser 290 295 300 Trp Glu Tyr Trp Gly Ala Gln Leu Asn Ala Met
Lys Pro Asp Leu Gln 305 310 315 320 Arg Ala Leu Gly Ala Thr Pro Asn
Thr Gly Pro Ala Pro Gln Gly Ala 325 330 335 <210> SEQ ID NO
71 <211> LENGTH: 1038 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polynucleotide <400> SEQUENCE: 71 tttgggccca ttatgtcgga
attcgcgtac ggttccttcg ttcgcacggt gtcgctgccg 60 gtaggtgctg
acgaggggaa tctattcatt gctccttggg gggttattca ccacccgcat 120
tatgaggaat gttcctgtta ctcccggccg ggcttgccgg tggagtacct gcaggtgccg
180 tcgccgtcga tgggccgtga catcaaggtc caattccaaa gtggtggtgc
caactcgccc 240 gccctgtacc tgctcgacgg cctgcgcgcg caggacgact
tcagcggctg ggacatcaac 300 accccggcgt tcgagtggta cgaccagtcg
ggcctgtcgg tggtcatgcc ggtgggtggc 360 cagtcaagct tctactccga
ctggtaccag cccgcctgcg gcaaggccgg ttgccagact 420 tacaagtggg
agaccttcct gaccagcgag ctgccggggt ggctgcaggc caacaggcac 480
gtcaagccca ccggaagcgc cgtcgtcggt ctttcgatgg ctgcttcttc ggcgctgacg
540 ctggcgatct atcaccccca gcagttcgtc tacgcgggag cgatgtcggg
cctgttggac 600 ccctcccagg cgatgggtcc caccctgatc ggcctggcga
tgggtgacgc tggcggctac 660 aaggcctccg acatgtgggg cccgaaggag
gacccggcgt ggcagcgcaa cgacccgctg 720 ttgaacgtcg ggaagctgat
cgccaacaac acccgcgtct gggtgtactg cggcaacggc 780 aagccgtcgg
atctgggtgg caacaacctg ccggccaagt tcctcgaggg cttcgtgcgg 840
accagcaaca tcaagttcca agacgcctac aacgccggtg gcggccacaa cggcgtgttc
900 gacttcccgg acagcggtac gcacagctgg gagtactggg gcgcgcagct
caacgctatg 960 aagcccgacc tgcaacgggc actgggtgcc acgcccaaca
ccgggcccgc gccccagggc 1020 gcctagtttc ttaagttt 1038 <210> SEQ
ID NO 72 <211> LENGTH: 21 <212> TYPE: DNA <213>
ORGANISM: Influenza virus <400> SEQUENCE: 72 gggaatctat
tcattgctcc t 21 <210> SEQ ID NO 73 <211> LENGTH: 21
<212> TYPE: DNA <213> ORGANISM: Influenza virus
<400> SEQUENCE: 73 tggggggtta ttcaccaccc g 21 <210> SEQ
ID NO 74 <211> LENGTH: 24 <212> TYPE: DNA <213>
ORGANISM: Influenza virus <400> SEQUENCE: 74 cattatgagg
aatgttcctg ttac 24 <210> SEQ ID NO 75 <211> LENGTH: 20
<212> TYPE: PRT <213> ORGANISM: Mycobacterium
tuberculosis <400> SEQUENCE: 75 Ser Glu Phe Ala Tyr Gly Ser
Phe Val Arg Thr Val Ser Leu Pro Val 1 5 10 15 Gly Ala Asp Glu 20
<210> SEQ ID NO 76 <211> LENGTH: 60 <212> TYPE:
DNA <213> ORGANISM: Mycobacterium tuberculosis <400>
SEQUENCE: 76 tcggaattcg cgtacggttc cttcgttcgc acggtgtcgc tgccggtagg
tgctgacgag 60 <210> SEQ ID NO 77 <211> LENGTH: 22
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 77 Gly
Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His His Pro His Tyr 1 5 10
15 Glu Glu Cys Ser Cys Tyr 20 <210> SEQ ID NO 78 <211>
LENGTH: 66 <212> TYPE: DNA <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic oligonucleotide
<400> SEQUENCE: 78 gggaatctat tcattgctcc ttggggggtt
attcaccacc cgcattatga ggaatgttcc 60 tgttac 66 <210> SEQ ID NO
79 <211> LENGTH: 1008 <212> TYPE: DNA <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
polynucleotide <400> SEQUENCE: 79 tcggaattcg cgtacggttc
cttcgttcgc acggtgtcgc tgccggtagg tgctgacgag 60 gggaatctat
tcattgctcc ttggggggtt attcaccacc cgcattatga ggaatgttcc 120
tgttactccc ggccgggctt gccggtggag tacctgcagg tgccgtcgcc gtcgatgggc
180 cgtgacatca aggtccaatt ccaaagtggt ggtgccaact cgcccgccct
gtacctgctc 240 gacggcctgc gcgcgcagga cgacttcagc ggctgggaca
tcaacacccc ggcgttcgag 300 tggtacgacc agtcgggcct gtcggtggtc
atgccggtgg gtggccagtc aagcttctac 360 tccgactggt accagcccgc
ctgcggcaag gccggttgcc agacttacaa gtgggagacc 420 ttcctgacca
gcgagctgcc ggggtggctg caggccaaca ggcacgtcaa gcccaccgga 480
agcgccgtcg tcggtctttc gatggctgct tcttcggcgc tgacgctggc gatctatcac
540 ccccagcagt tcgtctacgc gggagcgatg tcgggcctgt tggacccctc
ccaggcgatg 600 ggtcccaccc tgatcggcct ggcgatgggt gacgctggcg
gctacaaggc ctccgacatg 660 tggggcccga aggaggaccc ggcgtggcag
cgcaacgacc cgctgttgaa cgtcgggaag 720 ctgatcgcca acaacacccg
cgtctgggtg tactgcggca acggcaagcc gtcggatctg 780 ggtggcaaca
acctgccggc caagttcctc gagggcttcg tgcggaccag caacatcaag 840
ttccaagacg cctacaacgc cggtggcggc cacaacggcg tgttcgactt cccggacagc
900 ggtacgcaca gctgggagta ctggggcgcg cagctcaacg ctatgaagcc
cgacctgcaa 960 cgggcactgg gtgccacgcc caacaccggg cccgcgcccc agggcgcc
1008 <210> SEQ ID NO 80 <211> LENGTH: 10 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 80 Cys Ala Gly Ala Gly Asn
Phe Ile Ala Pro 1 5 10 <210> SEQ ID NO 81 <211> LENGTH:
10 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 81 Cys
Ala Gly Ala Gly Asn Leu Ile Ala Pro 1 5 10 <210> SEQ ID NO 82
<211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 82 Cys Ala Gly Ala Gly Asn Leu Phe Ile Ala
Pro 1 5 10 <210> SEQ ID NO 83 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 83 Cys
Ala Gly Ala Trp Gly Val His His Pro 1 5 10 <210> SEQ ID NO 84
<211> LENGTH: 10 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 84 Cys Ala Gly Ala Trp Gly Ile His His Pro 1
5 10 <210> SEQ ID NO 85 <211> LENGTH: 11 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 85 Cys Ala Gly
Ala Trp Gly Val Ile His His Pro 1 5 10 <210> SEQ ID NO 86
<211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 86 Cys Ala Gly Ala Trp Gly Ile Val His His
Pro 1 5 10 <210> SEQ ID NO 87 <211> LENGTH: 13
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 87 Gly
Asn Leu Ile Ala Pro Trp Gly Val Ile His His Pro 1 5 10 <210>
SEQ ID NO 88 <211> LENGTH: 17 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 88 Cys Ala Gly Ala Gly Asn
Leu Ile Ala Pro Trp Gly Val Ile His His 1 5 10 15 Pro <210>
SEQ ID NO 89 <211> LENGTH: 14 <212> TYPE: PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 89 Gly Asn Leu Phe Ile Ala
Pro Trp Gly Val Ile His His Pro 1 5 10 <210> SEQ ID NO 90
<211> LENGTH: 18 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 90 Cys Ala Gly Ala Gly Asn Leu Phe Ile Ala
Pro Trp Gly Val Ile His 1 5 10 15 His Pro <210> SEQ ID NO 91
<211> LENGTH: 8 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 91 His Tyr Glu Glu Cys Ser Cys Tyr 1 5
<210> SEQ ID NO 92 <211> LENGTH: 12 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 92 Cys Ala Gly Ala His Tyr
Glu Glu Cys Ser Cys Tyr 1 5 10 <210> SEQ ID NO 93 <211>
LENGTH: 22 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 93 Gly Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His His
Pro His Tyr 1 5 10 15 Glu Glu Cys Ser Cys Tyr 20 <210> SEQ ID
NO 94 <211> LENGTH: 26 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 94 Cys Ala Gly Ala Gly Asn Leu Phe
Ile Ala Pro Trp Gly Val Ile His 1 5 10 15 His Pro His Tyr Glu Glu
Cys Ser Cys Tyr 20 25 <210> SEQ ID NO 95 <211> LENGTH:
28 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 95 Gly
Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His His Pro Gly Asn 1 5 10
15 Leu Phe Ile Ala Pro Trp Gly Val Ile His His Pro 20 25
<210> SEQ ID NO 96 <211> LENGTH: 32 <212> TYPE:
PRT
<213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 96 Cys Ala Gly Ala Gly
Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His 1 5 10 15 His Pro Gly
Asn Leu Phe Ile Ala Pro Trp Gly Val Ile His His Pro 20 25 30
<210> SEQ ID NO 97 <211> LENGTH: 22 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 97 His Tyr Glu Glu Cys Ser
Cys Tyr Gly Asn Leu Phe Ile Ala Pro Trp 1 5 10 15 Gly Val Ile His
His Pro 20 <210> SEQ ID NO 98 <211> LENGTH: 22
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 98 Gly
Asn Leu Phe Ile Ala Pro His Tyr Glu Glu Cys Ser Cys Tyr Trp 1 5 10
15 Gly Val Ile His His Pro 20 <210> SEQ ID NO 99 <211>
LENGTH: 39 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic polypeptide
<400> SEQUENCE: 99 Ser Leu Leu Thr Glu Val Glu Thr Pro Ile
Arg Asn Glu Trp Gly Leu 1 5 10 15 Leu Thr Glu Val Glu Thr Pro Ile
Arg Gln Tyr Ile Lys Ala Asn Ser 20 25 30 Lys Phe Ile Gly Ile Thr
Glu 35 <210> SEQ ID NO 100 <211> LENGTH: 35 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic polypeptide <400> SEQUENCE: 100 Gly Asn
Leu Phe Ile Ala Pro Gly Asn Leu Phe Ile Ala Pro Gln Tyr 1 5 10 15
Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu Gly Asn Leu Phe 20
25 30 Ile Ala Pro 35 <210> SEQ ID NO 101 <211> LENGTH:
48 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic polypeptide <400> SEQUENCE:
101 His Tyr Glu Glu Cys Ser Cys Tyr Asp Trp Ser Gly Tyr Ser Gly Ser
1 5 10 15 Phe Val Gln His Pro Glu Leu Thr Gly Leu His Tyr Glu Glu
Cys Ser 20 25 30 Cys Tyr Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile
Gly Ile Thr Glu 35 40 45 <210> SEQ ID NO 102 <211>
LENGTH: 52 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic polypeptide
<400> SEQUENCE: 102 Val Thr Arg Glu Pro Tyr Val Ser Cys Asp
Pro Lys Ser Cys Ile Asn 1 5 10 15 Arg Cys Phe Tyr Val Glu Leu Ile
Arg Gly Arg Val Thr Arg Glu Pro 20 25 30 Tyr Val Ser Cys Asp Pro
Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile 35 40 45 Gly Ile Thr Glu 50
<210> SEQ ID NO 103 <211> LENGTH: 18 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 103 Asp Trp Ser Gly Tyr Ser
Gly Ser Phe Val Gln His Pro Glu Leu Thr 1 5 10 15 Gly Leu
<210> SEQ ID NO 104 <211> LENGTH: 33 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic polypeptide <400> SEQUENCE: 104 Ile Thr Gly Phe Ala
Pro Phe Ser Lys Asp Asn Ser Ile Arg Leu Ser 1 5 10 15 Ala Gly Gly
Asp Ile Trp Val Thr Arg Glu Pro Tyr Val Ser Cys Asp 20 25 30 Pro
<210> SEQ ID NO 105 <211> LENGTH: 16 <212> TYPE:
PRT <213> ORGANISM: Artificial Sequence <220> FEATURE:
<223> OTHER INFORMATION: Description of Artificial Sequence:
Synthetic peptide <400> SEQUENCE: 105 Lys Ser Cys Ile Asn Arg
Cys Phe Tyr Val Glu Leu Ile Arg Gly Arg 1 5 10 15 <210> SEQ
ID NO 106 <211> LENGTH: 12 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 106 Gly Asn Leu Phe Ile Ala Pro Arg
Tyr Ala Phe Ala 1 5 10 <210> SEQ ID NO 107 <211>
LENGTH: 16 <212> TYPE: PRT <213> ORGANISM: Artificial
Sequence <220> FEATURE: <223> OTHER INFORMATION:
Description of Artificial Sequence: Synthetic peptide <400>
SEQUENCE: 107 Cys Ala Gly Ala Gly Asn Leu Phe Ile Ala Pro Arg Tyr
Ala Phe Ala 1 5 10 15 <210> SEQ ID NO 108 <211> LENGTH:
11 <212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 108
Gly Asn Leu Val Val Pro Arg Tyr Ala Phe Ala 1 5 10 <210> SEQ
ID NO 109 <211> LENGTH: 15 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 109 Cys Ala Gly Ala Gly Asn Leu Val
Val Pro Arg Tyr Ala Phe Ala 1 5 10 15 <210> SEQ ID NO 110
<211> LENGTH: 11 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 110 Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe
Ala 1 5 10 <210> SEQ ID NO 111 <211> LENGTH: 15
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 111
Cys Ala Gly Ala Gly Asn Leu Ile Ala Pro Arg Tyr Ala Phe Ala 1 5 10
15 <210> SEQ ID NO 112 <211> LENGTH: 6 <212>
TYPE: PRT <213> ORGANISM: Artificial Sequence <220>
FEATURE: <223> OTHER INFORMATION: Description of Artificial
Sequence: Synthetic peptide <400> SEQUENCE: 112 Gly Asn Leu
Val Val Pro 1 5 <210> SEQ ID NO 113 <211> LENGTH: 10
<212> TYPE: PRT <213> ORGANISM: Artificial Sequence
<220> FEATURE: <223> OTHER INFORMATION: Description of
Artificial Sequence: Synthetic peptide <400> SEQUENCE: 113
Cys Ala Gly Ala Gly Asn Leu Val Val Pro 1 5 10 <210> SEQ ID
NO 114 <211> LENGTH: 15 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 114 Phe Val Ile Arg Glu Pro Phe Ile
Ser Cys Ser His Leu Glu Cys 1 5 10 15 <210> SEQ ID NO 115
<211> LENGTH: 19 <212> TYPE: PRT <213> ORGANISM:
Artificial Sequence <220> FEATURE: <223> OTHER
INFORMATION: Description of Artificial Sequence: Synthetic peptide
<400> SEQUENCE: 115 Cys Ala Gly Ala Phe Val Ile Arg Glu Pro
Phe Ile Ser Cys Ser His 1 5 10 15 Leu Glu Cys <210> SEQ ID NO
116 <211> LENGTH: 4 <212> TYPE: PRT <213>
ORGANISM: Artificial Sequence <220> FEATURE: <223>
OTHER INFORMATION: Description of Artificial Sequence: Synthetic
peptide <400> SEQUENCE: 116 Cys Ala Gly Ala 1
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