U.S. patent application number 12/784296 was filed with the patent office on 2011-02-24 for vaccine against varicella zoster virus.
This patent application is currently assigned to GLAXOSMITHKLINE BIOLOGICALS, S.A.. Invention is credited to EMMANUEL JULES HANON, JEAN STEPHENNE.
Application Number | 20110045059 12/784296 |
Document ID | / |
Family ID | 34430592 |
Filed Date | 2011-02-24 |
United States Patent
Application |
20110045059 |
Kind Code |
A1 |
HANON; EMMANUEL JULES ; et
al. |
February 24, 2011 |
VACCINE AGAINST VARICELLA ZOSTER VIRUS
Abstract
Use of an immunogenic composition comprising VZV gE, or
immunogenic fragment thereof, and a TH-1 adjuvant in the
preparation of a medicament for the prevention or amelioration of
shingles and/or post herpetic neuralgia. Compositions comprising a
truncated VZV gE antigen and an adjuvant containing QS21,
cholesterol and 3D MPL are also claimed
Inventors: |
HANON; EMMANUEL JULES;
(RIXENSART, BE) ; STEPHENNE; JEAN; (RIXENSART,
BE) |
Correspondence
Address: |
Convergent Law Group LLP
P.O. BOX 1329
MOUNTAIN VIEW
CA
94042
US
|
Assignee: |
GLAXOSMITHKLINE BIOLOGICALS,
S.A.
RIXENSART
BE
|
Family ID: |
34430592 |
Appl. No.: |
12/784296 |
Filed: |
May 20, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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11817175 |
Aug 27, 2007 |
|
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12784296 |
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Current U.S.
Class: |
424/450 ;
424/186.1; 424/230.1 |
Current CPC
Class: |
A61P 31/22 20180101;
A61P 29/00 20180101; A61P 31/00 20180101; A61P 37/04 20180101; C12N
2710/16734 20130101; A61K 2039/5252 20130101; A61P 31/18 20180101;
A61K 2039/55555 20130101; A61K 39/25 20130101; C07K 14/005
20130101; A61K 2039/55577 20130101; A61P 25/00 20180101; A61P 25/04
20180101; A61K 2039/5254 20130101; A61K 39/12 20130101; A61K
2039/55572 20130101; C12N 2710/16722 20130101; A61K 39/00
20130101 |
Class at
Publication: |
424/450 ;
424/230.1; 424/186.1 |
International
Class: |
A61K 39/25 20060101
A61K039/25; A61K 9/127 20060101 A61K009/127; A61P 31/22 20060101
A61P031/22; A61P 37/04 20060101 A61P037/04; A61P 25/00 20060101
A61P025/00 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 3, 2005 |
GB |
0504436.7 |
Mar 1, 2006 |
EP |
PCT/EP2006/002070 |
Claims
1. A method of preventing herpes zoster reactivation in an
individual, said method comprising sequential delivery of: a live
attenuated or whole inactivated varicella-zoster virus (VZV); and
an immunogenic composition comprising VZV gE, 3D-MPL and QS21.
2. The method of claim 1, wherein 3D-MPL is provided in
liposomes.
3. The method of claim 2, wherein the liposomes comprise
cholesterol.
4. The method of claim 1, wherein two or more immunizations are
employed.
5. The method of claim 4, wherein the immunogenic composition is
delivered first, followed by delivery of the live attenuated or
whole inactivated VZV.
6. The method of claim 1, wherein the live attenuated or whole
inactivated VZV is delivered first, followed by delivery of the
immunogenic composition.
7. The method of claim 1, wherein the live attenuated or whole
inactivated VZV and the immunogenic composition are administered in
a two dose regime comprising between 1-6 months between individual
immunizations.
8. The method of claim 7, wherein the immunogenic composition is
administered between 1-6 months following the delivery of the live
attenuated or whole inactivated VZV.
9. The method of claim 7, wherein the live attenuated or whole
inactivated VZV is delivered between 1-6 months following the
delivery of the immunogenic composition.
10. The method of claim 1, wherein the VZV is a live attenuated OKA
strain.
11. The method according to claim 1, wherein the gE is a
truncate.
12. The method according to claim 11, wherein the gE is a
C-terminal truncate.
13. The method according to claim 12, wherein the gE has the amino
acid sequence of SEQ ID NO: 1.
14. The method of claim 1, wherein between 25-100 .mu.g of VZV gE
are delivered.
15. The method of claim 14, wherein between 40-100 .mu.g of VZV gE
are delivered.
16. The method of claim 1, further comprising repeating the
delivery of the live attenuated or whole inactivated VZV.
17. The method of claim 16, comprising repeating the delivery of
the live attenuated or whole inactivated VZV two or more times.
18. The method according to claim 1, wherein the sequential
delivery is administered to an individual of at least 50 years of
age.
19. The method according to claim 1, wherein the sequential
delivery is administered to an immunocompromised individual.
20. A method of ameliorating the severity of herpes zoster
reactivation and/or post herpetic neuralgia in an individual, said
method comprising sequential delivery of a live attenuated or whole
inactivated varicella-zoster virus (VZV); and an immunogenic
composition comprising VZV gE, 3D-MPL and QS21.
21. The method of claim 20, wherein 3D-MPL is provided in
liposomes.
22. The method of claim 21, wherein the liposomes comprise
cholesterol.
23. The method of claim 20, wherein two or more immunizations are
employed.
24. The method of claim 23, wherein the immunogenic composition is
delivered first, followed by delivery of the live attenuated or
whole inactivated VZV.
25. The method of claim 20, wherein the live attenuated or whole
inactivated VZV is delivered first, followed by delivery of the
immunogenic composition.
26. The method of claim 20, wherein the live attenuated or whole
inactivated VZV and the immunogenic composition are administered in
a two dose regime comprising between 1-6 months between doses.
27. The method of claim 26, wherein the immunogenic composition is
administered between 1-6 months following the delivery of the live
attenuated or whole inactivated VZV.
28. The method of claim 26, wherein the live attenuated or whole
inactivated VZV is delivered between 1-6 months following the
delivery of the immunogenic composition.
29. The method of claim 20, wherein the VZV is a live attenuated
OKA strain.
30. The method according to claim 20, wherein the gE is a
truncate.
31. The method according to claim 30, wherein the gE is a
C-terminal truncate.
32. The method according to claim 31, wherein the gE has the amino
acid sequence of SEQ ID NO: 1.
33. The method of claim 20, wherein between 25-100 .mu.g of VZV gE
are delivered.
34. The method of claim 33, wherein between 40-100 .mu.g of VZV gE
are delivered.
35. The method of claim 20, further comprising repeating the
delivery of the live attenuated or whole inactivated VZV.
36. The method of claim 35, comprising repeating the delivery of
the live attenuated or whole inactivated VZV two or more times.
37. The method according to claim 20, wherein the sequential
delivery is administered to an individual of at least 50 years of
age.
38. The method according to claim 1, wherein the sequential
delivery is administered to an immunocompromised individual.
39. A method of preventing herpes zoster reactivation in an
individual, said method comprising concomitant delivery of a live
attenuated or whole inactivated varicella-zoster virus (VZV); and
an immunogenic composition comprising VZV gE, 3D-MPL and QS21.
40. A method of ameliorating the severity of herpes zoster and/or
post herpetic neuralgia in an individual, said method comprising a
concomitant delivery of a live attenuated or whole inactivated
varicella-zoster virus (VZV); and an immunogenic composition
comprising VZV gE, 3D-MPL and QS21.
Description
[0001] This application claims priority from co-pending U.S.
application Ser. No. 11/817,175, filed Mar. 1, 2006, which claims
priority to GB application 0504436.7, filed Mar. 3, 2005, each of
which is hereby incorporated by reference in its entirety.
[0002] This invention relates to compositions capable of inducing
an immune response against Varicella-Zoster Virus.
[0003] Varicella-Zoster Virus (VZV) is a human herpes virus which
is the etiological agent of chicken pox (varicella) and shingles
(zoster). Varicella results from an initial, or primary infection,
usually contracted during childhood which is relatively benign.
However, for adults who were not exposed to varicella during
childhood, and occasionally to individuals who are immunocomprised,
VZV can be life-threatening. Similarly, a VZV infection can be
life-threatening to neonates, for the virus is capable of crossing
the placenta. With direct contact, varicella is known to be a
highly transmissible infectious disease.
[0004] Like most Herpes-Viruses, VZV has a tendency to infect some
cells in which its development is arrested. After a variable latent
period, the Varicella-Zoster (VZ) virus can be released to initiate
infection in other cells. This reactivation of the VZ virus causes
an estimated 5 million cases of zoster annually (Plotkin et al.,
Postgrad Med J 61: 155-63 (1985)). Zoster is characterized by
inflammation of the cerebral ganglia and peripheral nerves, and it
is associated with acute pain.
[0005] It has been shown that humans vaccinated with attenuated
strains of VZV have received protective immunity from VZV
infections (Arbeter et al., J. Pediatr 100 886-93 (1982) and
Brunell et al., Lancet ii: 1069-72 (1982)). In particular the OKA
strain of VZV has been used in trials for the prevention of herpes
zoster and post herpetic neuralgia. The OKA strain has also been
used in the preparation of vaccines for chickenpox for many years
and is well characterised--for example see EP651789 and references
therein.
[0006] A large clinical trial using the OKA strain for the zoster
indication has been published in The New England Journal of
Medicine 2005, number 22, Volume 352:2271-2284 (M. N. Oxman et
al).
[0007] There is still a need for improved vaccines against herpes
zoster and related disorders such as post herpetic neuralgia
(PHN).
STATEMENT OF INVENTION
First Aspect
[0008] The present invention provides in a first aspect an
immunogenic composition comprising a VZV antigen or immunogenic
derivative thereof in combination with a live attenuated VZV or
whole inactivated VZV.
[0009] The invention further relates to a vaccine composition
comprising a VZV antigen or immunogenic derivative thereof in
combination with a live attenuated VZV or whole inactivated
VZV.
[0010] The invention further relates to a method of preventing
and/or decreasing the severity of herpes zoster and/or post
herpetic neuralgia (PHN) comprising delivering to an individual an
immunogenic composition comprising a VZV antigen or immunogenic
derivative thereof in combination with a live attenuated VZV or
whole inactivated VZV.
[0011] In a further embodiment the invention relates to a method of
preventing and/or decreasing the severity of herpes zoster and/or
post herpetic neuralgia, the method comprising sequential or
concomitant delivery to an individual of a VZV antigen or
immunogenic derivative thereof and a live attenuated VZV or whole
inactivated VZV.
[0012] In a still further embodiment the invention relates to a kit
comprising a live attenuated VZV or whole inactivated VZV and,
separately, a VZV antigen or immunogenic derivative thereof, the
components suitable for concomitant or sequential delivery, or for
mixing as a single composition prior to delivery.
[0013] The invention also relates to a method for the manufacture
of an immunogenic composition, the method comprising combining a
live attenuated VZV or whole inactivated VZV with a VZV antigen or
immunogenic derivative thereof.
[0014] The invention further relates to use of a live attenuated
VZV strain or whole inactivated VZV and a VZV antigen or
immunogenic derivative thereof in the preparation of an immunogenic
composition for preventing and/or decreasing the severity of herpes
zoster and/or post herpetic neuralgia.
Second Aspect
[0015] In a second aspect the invention relates to an immunogenic
composition and/or vaccine comprising gE or an immunogenic
derivative or immunogenic fragment thereof in combination with a
TH1-adjuvant.
[0016] The invention also relates to use of a composition
comprising gE or an immunogenic derivative or immunogenic fragment
thereof in combination with a TH1-adjuvant, in the preparation of a
medicament for the prevention or amelioration of herpes zoster
reactivation and/or post herpetic neuralgia.
[0017] The invention also relates to a method for the prevention or
amelioration of herpes zoster reactivation and/or post herpetic
neuralgia, the method comprising delivering to an individual in
need thereof an immunogenic composition or vaccine comprising gE or
an immunogenic derivative or immunogenic fragment thereof in
combination with a TH1-adjuvant.
FIGURES
[0018] FIG. 1 discloses the sequence of a truncated VZV gE.
[0019] FIGS. 2-4 disclose humoral responses obtained in human
clinical trials using compositions of the invention.
[0020] FIGS. 5 and 6 disclose cell mediated immunity obtained in
human clinical trials using compositions of the invention.
DETAILED DESCRIPTION
[0021] In its broadest aspect the present invention relates to
compositions and regimes as described herein for provoking an
immune response to VZV. In one aspect the immune response generated
by exposure to such compositions is suitably reproducibly higher
and statistically significant when compared to that obtained in
individuals who have received no exposure to the compositions of
the invention. The immune response may be assessed by analysis of
any one or more aspects of CMI response and/or antibody responses
using any of the techniques outlined below.
[0022] In another aspect the invention relates to approaches for
preventing and/or decreasing the severity of herpes zoster and/or
post herpetic neuralgia (PHN). For the avoidance of doubt, the
invention relates in one aspect to use in the prevention of the
incidence of zoster. Where zoster does occur then the severity of
the reactivation of zoster is suitably reduced compared with an
unvaccinated control (amelioration of zoster). In a further aspect,
where zoster does occur, the invention relates to use in the
prevention of the incidence of PHN. In a further aspect where PHN
does occur then the severity of the PHN is suitably reduced
compared with an unvaccinated control (amelioration of PHN).
Reduction in severity can suitably be assessed by a reduction in
the pain caused by zoster or PHN, for example, using known measures
of burden of pain (e.g. Coplan et al J Pain 2004; 5 (6) 344-56).
Reduction in severity can also be assessed by other criteria such
as duration of zoster or PHN, proportion of body area affected by
zoster or PHN; or the site of zoster/PHN.
[0023] The above statements relate to all aspects of the
invention.
[0024] Where a live attenuated strain is used in the first aspect
of the invention, then in one aspect the live attenuated VZV strain
is the OKA strain, a strain well known in the art, for example as
disclosed in Arbeter et al. (Journal of Pediatrics, vol 100, No 6,
p 886 ff), WO9402596, and references therein, such as U.S. Pat. No.
3,985,615, all incorporated herein by reference. Any other suitable
live attenuated strain may also be used in the invention. For
example, the Varilrix.TM. and Varivax.TM. strains are both
appropriate and commercially available and could be employed in the
invention.
[0025] Whole inactivated VZV strains, such as inactivated VZV OKA
are also suitable for use in the present invention.
[0026] The VZV antigen for use in the invention may be any suitable
VZV antigen or immunogenic derivative thereof, suitably being a
purified VZV antigen.
[0027] In one aspect the antigen or derivative is one that is able
to elicit, when delivered in combination, concomitantly or
sequentially with a live attenuated VZV strain or whole inactivated
VZV, an immune response which is improved over that elicited by the
live attenuated strain/whole inactivated strain alone or by the VZV
antigen alone. Such a response may be, for example, improved in
terms of one or more of the magnitude of immune response, duration
of immune response, the number or % or responders, or the breadth
of response (e.g. the range of antibody or T cell responses
detected), or may provide an improvement at the clinical level in
terms of incidence, reduction of pain or symptoms. Improvements in
the immune response can be assessed by, for example, antibody
levels or cell mediated immunity (CMI) activity using standard
techniques in the art; improvements at the clinical level can be
also assessed using known clinical criteria.
[0028] In particular, in one aspect the immune response elicited by
the composition or vaccine of present invention shows one or more
of:
[0029] a statistically significant increase in the CMI and/or
antibody response, in comparison with pre-vaccination levels, when
compared to VZV antigen or live attenuated strain/whole inactivated
strain alone;
[0030] An improved multivalent CMI response, in comparison with
pre-vaccination levels, when compared to VZV antigen or live
attenuated strain/whole inactivated strain alone. A multivalent CMI
response considers a range of markers for CMI such as (but not
limited to) IFN gamma, IL2, TNF alpha and CD40L and an improved
multivalent response induces a CMI response across a wider range of
such markers or a higher response in one or more of the markers
when compared to a VZV antigen or live attenuated strain/whole
inactivated strain alone;
[0031] Better persistent CMI or antibody response, in comparison
with pre-vaccination levels, when compared to VZV antigen or live
attenuated strain/whole inactivated strain alone. In one aspect
persistence is measured over after 1 month, 2 months, 3 months, 4
months, 5 months, 6 months, 12 months, 24 months, 36 months or 48
months.
[0032] In one aspect improvements in the immune response are
assessed in the elderly population, suitably the populations over
50 years of age, for whom the risk of zoster or PHN is increased
with respect to the population under 50 years of age. Improved
immune responses can also be examined in immuno-compromised
populations. In one aspect such populations are target populations
for any embodiment of the present invention.
[0033] In one aspect the population is over 50 years, suitably over
60 years, over 70 years, or even over 80 years and above. In one
aspect the population is 50-70 years old.
[0034] Thus in one aspect the invention relates to use of the
compositions and approaches of the invention in preventing and/or
decreasing the severity of zoster or PHN in humans over 50 years of
age.
[0035] In one aspect the invention relates to use of the
compositions and approaches of the invention in preventing and/or
decreasing the severity of zoster or PHN in immunocompromised
individuals, such as transplant patients or those who are HIV
positive.
[0036] The term `immunogenic derivative` encompasses any molecule
which retains the ability to induce an immune response to VZV
following administration to man. Immunogenic compounds herein are
suitably capable of reacting detectably within an immunoassay (such
as an ELISA or T-cell stimulation assay) with antisera and/or
T-cells from a patient with VZV. Screening for immunogenic activity
can be performed using techniques well known to the skilled
artisan. For example, such screens can be performed using methods
such as those described in Harlow and Lane, Antibodies: A
Laboratory Manual, Cold Spring Harbor Laboratory, 1988.
[0037] Suitable methods for the generation of derivatives are well
known in the art and include standard molecular biology techniques
as disclosed, for example, in Sambrook et al [Molecular Cloning: A
Laboratory Manual, third edition, 2000, Cold Spring Harbor
Laboratory Press], such as techniques for the addition, deletion,
substitution or rearrangement of amino acids or chemical
modifications thereof. In one aspect derivatives include, for
example, truncations or other fragments.
[0038] In one aspect derivatives in the context of this invention
are amino acid sequences comprising epitopes, i.e., antigenic
determinants substantially responsible for the immunogenic
properties of a polypeptide and being capable of eliciting an
immune response, in one aspect being T cell epitopes.
[0039] In one aspect, the level of immunogenic activity of the
immunogenic derivative is at least about 50%, in one aspect at
least about 70% and in one aspect at least or greater than about
90% of the immunogenicity for the polypeptide from which it is
derived, suitably as assessed by immunoassay techniques described
above. In some aspects of the invention immunogenic portions may be
identified that have a level of immunogenic activity greater than
that of the corresponding full-length polypeptide, e.g., having
greater than about 100% or 150% or more immunogenic activity.
[0040] In one aspect the VZV antigen is a glycoprotein, in one
aspect the gE antigen (also known as gp1), or immunogenic
derivative thereof.
[0041] The gE antigen, anchorless derivatives thereof (which are
also immunogenic derivatives) and production thereof is described
in EPO405867 and references therein [see also Vafai A. Antibody
binding sites on truncated forms of varicella-zoster virus gpI(gE)
glycoprotein Vaccine 1994 12:1265-9]. EP192902 also discloses gE
and production thereof.
[0042] The disclosure of all cited documents is herein fully
incorporated by reference.
[0043] In one aspect gE is a truncated gE having the sequence of
FIG. 1 herein, and as disclosed in Virus research, vol 40, 1996 p
199 ff, herein incorporated fully by reference. Reference to gE
hereinafter includes reference to truncated gE, unless otherwise
apparent from the context.
[0044] Other suitable antigens include, by way of example, gB, gH,
gC, gI, IE63 (e.g. see, Huang et al. J. Virol. 1992, 66: 2664,
Sharp et al. J. Inf. Dis. 1992, 165:852, Debrus, J. Virol. 1995
May; 69(5):3240-5 and references therein), IE62 (e.g. see Arvin et
al. J. Immunol. 1991 146:257, Sabella J. Virol. 1993 December;
67(12):7673-6 and references therein) ORF4 or ORF 10 (Arvin et al.
Viral Immunol. 2002 15: 507.)
[0045] The present invention herein also contemplates that antigen
combinations may be used with the live attenuated or killed VZV,
and in one aspect gE may be included in any such combination. In
one aspect the invention relates to combinations of gE with IE63
and gE with IE62, for example.
[0046] VZV antigens and derivatives of VZV antigens can be tested
for suitable immunogenic activity by use in the model systems as
described in the Examples of the present application, or by
clinical trials in humans. One or more of the following indicators
of activity are suitable for consideration in assessment of
immunogenic activity:
[0047] Increased CD4 or CD8 T cell responses to VZV or antigen
derivatives.
[0048] Elevation in VZV or antigens derivative specific
antibodies.
[0049] Enhanced production of cytokines such as interferon .gamma.
or IL-2 or TNF .alpha..
[0050] Enhanced expression of CD40L on CD4 and CD8 T cells.
[0051] Reduction in the incidence of zoster below the incidence
found in the general population of similarly at risk individuals,
and likewise reduced disease severity and/or associated pain below
the incidence found in the general population of similarly at risk
individuals.
[0052] Increases or reductions, as described above, are suitably
statistically significant with respect to appropriate controls,
such as an age-matched non-vaccinated group. In one aspect the live
attenuated VZV or killed VZV and the VZV antigen or antigen
derivatives do not significantly interfere with one another, such
that when used in combination the 2 components are still able to
provide an immunogenic response to VZV. In one aspect the response
is a protective immunogenic response, whether the 2 components are
used as a composition or used in sequential administration or
coadministration. It will be appreciated that some interference is
tolerated, however, provided that the overall protective immune
response is improved in some way (increased in magnitude, increased
% responders or broadened antigenic responses, for example) over
that of either of the original components used individually.
[0053] In one aspect the invention relates to the combination of
the gE antigen and the OKA strain, used for concomitant or
sequential administration, in either order. Where delivery is
concomitant then the 2 components are delivered into different
injection sites but during the same day, for example. In one aspect
different delivery routes are used for the 2 components, in
particular subcutaneous delivery for the virus strain such as OKA
and intramuscular delivery for the antigen such as gE
[0054] The present invention also extends to cover, in all
embodiments described, the use of combinations of VZV antigens or
derivatives with a live attenuated VZV strain or killed VZV.
Suitable combinations of antigens include in one aspect gE or
immunogenic derivative thereof.
[0055] The combined composition, or either or both of the
individual components may additionally comprise an adjuvant or
immunostimulant such as but not limited to detoxified lipid A from
any source and non-toxic derivatives of lipid A, saponins and other
reagents, suitably capable of stimulating a TH1 type response.
[0056] In one aspect the composition comprises an adjuvant capable
of stimulating a TH1 type response.
[0057] High levels of Th1-type cytokines tend to favour the
induction of cell mediated immune responses to a given antigen,
whilst high levels of Th2-type cytokines tend to favour the
induction of humoral immune responses to the antigen.
[0058] The distinction of Th1 and Th2-type immune response is not
absolute. In reality an individual will support an immune response
which is described as being predominantly Th1 or predominantly Th2.
However, it is often convenient to consider the families of
cytokines in terms of that described in murine CD4+ ve T cell
clones by Mosmann and Coffman (Mosmann, T. R. and Coffman, R. L.
(1989) TH1 and TH2 cells: different patterns of lymphokine
secretion lead to different functional properties. Annual Review of
Immunology, 7, p 145-173). Traditionally, Th1-type responses are
associated with the production of the INF-.gamma. and IL-2
cytokines by T-lymphocytes. Other cytokines often directly
associated with the induction of Th1-type immune responses are not
produced by T-cells, such as IL-12. In contrast, Th2-type responses
are associated with the secretion of 11-4, IL-5, IL-6, IL-10.
[0059] Suitable adjuvant systems which promote a predominantly Th1
response include, Monophosphoryl lipid A or a derivative thereof,
particularly 3-de-O-acylated monophosphoryl lipid A. It has long
been known that enterobacterial lipopolysaccharide (LPS) is a
potent stimulator of the immune system, although its use in
adjuvants has been curtailed by its toxic effects. A non-toxic
derivative of LPS, monophosphoryl lipid A (MPL), produced by
removal of the core carbohydrate group and the phosphate from the
reducing-end glucosamine, has been described by Ribi et al (1986,
Immunology and Immunopharmacology of bacterial endotoxins, Plenum
Publ. Corp., NY, p 407-419) and has the following structure:
##STR00001##
[0060] A further detoxified version of MPL results from the removal
of the acyl chain from the 3-position of the disaccharide backbone,
and is called 3-O-Deacylated monophosphoryl lipid A (3D-MPL). It
can be purified and prepared by the methods taught in GB 2122204B,
which reference also discloses the preparation of diphosphoryl
lipid A, and 3-O-deacylated variants thereof.
[0061] In one aspect 3D-MPL is in the form of an emulsion having a
small particle size less than 0.2 .mu.m in diameter, and its method
of manufacture is disclosed in WO 94/21292. Aqueous formulations
comprising monophosphoryl lipid A and a surfactant have been
described in WO9843670A2.
[0062] The bacterial lipopolysaccharide derived adjuvants to be
formulated in the compositions of the present invention may be
purified and processed from bacterial sources, or alternatively
they may be synthetic. For example, purified monophosphoryl lipid A
is described in Ribi et al 1986 (supra), and 3-O-Deacylated
monophosphoryl or diphosphoryl lipid A derived from Salmonella sp.
is described in GB 2220211 and U.S. Pat. No. 4,912,094. Other
purified and synthetic lipopolysaccharides have been described
(Hilgers et al., 1986, Int. Arch. Allergy. Immunol., 79(4):392-6;
Hilgers et al., 1987, Immunology, 60(1):141-6; and EP 0 549 074
B1). In one aspect the bacterial lipopolysaccharide adjuvant is
3D-MPL.
[0063] Accordingly, the LPS derivatives that may be used in the
present invention are those immuno stimulants that are similar in
structure to that of LPS or MPL or 3D-MPL. In another embodiment of
the present invention the LPS derivatives may be an acylated
monosaccharide, which is a sub-portion to the above structure of
MPL.
[0064] Saponins are taught in: Lacaille-Dubois, M and Wagner H.
(1996. A review of the biological and pharmacological activities of
saponins. Phytomedicine vol 2 pp 363-386). Saponins are steroid or
triterpene glycosides widely distributed in the plant and marine
animal kingdoms. Saponins are noted for forming colloidal solutions
in water which foam on shaking, and for precipitating cholesterol.
When saponins are near cell membranes they create pore-like
structures in the membrane which cause the membrane to burst.
Haemolysis of erythrocytes is an example of this phenomenon, which
is a property of certain, but not all, saponins.
[0065] Saponins are known as adjuvants in vaccines for systemic
administration. The adjuvant and haemolytic activity of individual
saponins has been extensively studied in the art (Lacaille-Dubois
and Wagner, supra). For example, Quil A (derived from the bark of
the South American tree Quillaja Saponaria Molina), and fractions
thereof, are described in U.S. Pat. No. 5,057,540 and "Saponins as
vaccine adjuvants", Kensil, C. R., Crit. Rev Ther Drug Carrier
Syst, 1996, 12 (1-2):1-55; and EP 0 362 279 B1. Particulate
structures, termed Immune Stimulating Complexes (ISCOMS),
comprising fractions of Quil A are haemolytic and have been used in
the manufacture of vaccines (Morein, B., EP 0 109 942 B1; WO
96/11711; WO 96/33739). The haemolytic saponins QS21 and QS17 (HPLC
purified fractions of Quil A) have been described as potent
systemic adjuvants, and the method of their production is disclosed
in U.S. Pat. No. 5,057,540 and EP 0 362 279 B1. Other saponins
which have been used in systemic vaccination studies include those
derived from other plant species such as Gypsophila and Saponaria
(Bomford et al., Vaccine, 10(9):572-577, 1992).
[0066] An enhanced system involves the combination of a non-toxic
lipid A derivative and a saponin derivative particularly the
combination of QS21 and 3D-MPL as disclosed in WO 94/00153, or a
less reactogenic composition where the QS21 is quenched with
cholesterol as disclosed in WO 96/33739. In one aspect the
combination of QS21 with 3D MPL is used in the present
invention.
[0067] In one aspect the adjuvant for use in the invention
comprises QS21 and a liposomal formulation comprising cholesterol
and 3D MPL.
[0068] A particularly potent adjuvant formulation involving QS21
and 3D-MPL in an oil in water emulsion is described in WO 95/17210
and is also suitable for use in the present invention.
[0069] Accordingly in one embodiment of the present invention there
is provided a composition comprising a VZV antigen or derivative of
the present invention adjuvanted with detoxified lipid A or a
non-toxic derivative of lipid A. In one aspect the composition is
adjuvanted with a monophosphoryl lipid A or derivative thereof.
[0070] In one aspect the composition additionally comprises a
saponin, which in one aspect is QS21, and in another aspect is QS21
quenched with cholesterol as disclosed in WO 96/33739.
[0071] The immunogenic composition of the invention optionally
comprises an oil in water emulsion, which may be used in
combination with other adjuvants such as QS21 and/or 3D MPL as
disclosed above. Adjuvant formulations comprising an oil in water
emulsion are disclosed in WO9911241 and WO9912565, incorporated
herein by reference.
[0072] An alternative adjuvant choice is an unmethylated CpG
dinucleotides ("CpG"). CpG is an abbreviation for
cytosine-guanosine dinucleotide motifs present in nucleic acid. CpG
oligonucleotides are disclosed in WO 96/02555 and EP 468520.
[0073] In one aspect a combination of any of the adjuvants of the
invention described herein (QS21 or QS21 quenched with
cholesterol+3DMPL, optionally with an oil in water emulsion) is
used with gE, or immunogenic derivative thereof, used in
concomitant or sequential administration with a live attenuated VZV
or inactivated whole VZV.
[0074] The present invention also provides a method for producing a
kit suitable for inducing an immune response against zoster, the
method comprising mixing a VZV antigen preparation of the present
invention together with an adjuvant or adjuvant combination, and
combining in a kit with a live attenuated VZV.
[0075] The amount of VZV antigen is selected as an amount which
induces an immunoprotective response without significant, adverse
side effects in typical vaccines. Such amount will vary depending
upon which specific immunogen is employed and how it is presented.
Generally, it is expected that each dose will comprise 1-1000 .mu.g
of protein, such as 2-100 .mu.g, or 5-60 .mu.g. Where gE is used
then in one aspect 25-100 .mu.g of gE may be used in humans, such
as 40-100 .mu.g of gE for human use, in one aspect about 25 .mu.g,
about 50 .mu.g or about 100 .mu.g of gE, suitably 25 .mu.g, 50
.mu.g or 100 .mu.g gE. For the OKA strain, for example, a suitable
dose is 500-50000 pfu/0.5 ml, such as 2000-6000 pfu/0.5 ml, with a
suitable dose of the GSK Varilrix Oka strain for example being
6000-25,000 per dose, for example 10,000 pfu/dose. Higher doses
such as 30,000 pfu, 40000 pfu, 50,000 pfu 60,000 pfu, 70000 pfu,
80000 pfu, 90000 pfu or even 100000 pfu may be employed.
[0076] An optimal amount for a particular vaccine can be
ascertained by standard studies involving observation of
appropriate immune responses in subjects. Following an initial
vaccination, subjects may receive one or several booster
immunisation adequately spaced.
[0077] The composition(s) of the present invention may be
formulated for any appropriate manner of administration, including
for example, topical, oral, nasal, mucosal, intravenous,
intradermal, intraperitoneal, subcutaneous and intramuscular
administration. Delivery of the OKA strain is, in one aspect, by
subcutaneous delivery.
[0078] The immunogenic composition of the present invention may be
used in a vaccine composition, optionally in combination with an
adjuvant and/or (other) suitable carrier.
[0079] The VZV antigen and attenuated VZV of the present invention
may be used together in a composition to provoke an immune response
to VZV, or separately--either concomitantly or sequentially in a
prime boost regime. For concomitant or sequential delivery the
components of the vaccine may be used in either order. In one
embodiment, delivery of a live attenuated VZV or whole inactivated
VZV is followed by a VZV antigen or immunogenic derivative thereof.
In another embodiment delivery of a VZV antigen or immunogenic
derivative thereof is followed by delivery of live attenuated VZV
or whole inactivated VZV.
[0080] The invention further relates to a method of preventing
and/or decreasing the severity of herpes zoster and/or post
herpetic neuralgia comprising delivering to an individual at risk
of zoster an immunogenic composition comprising a live attenuated
VZV and a VZV antigen.
[0081] In a further embodiment the invention relates to a method of
preventing and/or decreasing the severity of herpes zoster and/or
post herpetic neuralgia comprising sequential or concomitant
delivery to an individual at risk of zoster of a live attenuated
VZV and a VZV antigen.
[0082] In one aspect the invention relates to a prime boost regime
wherein a VZV antigen, in one aspect an adjuvanted antigen, is
delivered first, after which the immune system is boosted with
delivery of an attenuated VZV.
[0083] A prime boost regime in humans comprises, in one aspect,
priming with 25-100 .mu.g gE, in one aspect 40-100 .mu.g gE, such
as 50 or about 50 .mu.g gE, or an immunogenic derivative thereof,
adjuvanted with QS21 (for example QS21 quenched with cholesterol as
described above) and 3D-MPL, and boosting with the OKA strain of
VZV.
[0084] Where prime boost regimes are used, or where multiple
vaccination regimes are used, then 2, 3, 4 or more immunisations
may be employed. Suitable regimes for prime boost include 1, 2, 3,
4, 5 or 6 months between individual immunisations.
[0085] A prime boost schedule comprises, in one aspect, delivery of
a VZV antigen or immunogenic derivative thereof, suitably an
adjuvanted VZV antigen or derivative, at 0 months and boosting with
a live attenuated VZV at 2M.
[0086] In an alternative delivery schedule there is concomitant
delivery of both of the two individual components (VZV antigen or
derivative and live attenuated VZV) at both 0 and 2 months.
[0087] In a still further embodiment the invention relates to a kit
comprising a live attenuated VZV or inactivated whole VZV and a VZV
antigen.
[0088] The invention also relates to a method for the manufacture
of an immunogenic composition, the method comprising combining a
live attenuated VZV/whole inactivated and a VZV antigen.
[0089] The invention further relates to use of a live attenuated
VZV strain in the preparation of a combination vaccine with a VZV
antigen for the prevention of herpes zoster, and to use of a VZV
antigen in the preparation of a combination vaccine with a live
attenuated VZV strain for the prevention of herpes zoster.
[0090] In a second, aspect of the invention a gE antigen, or
immunogenic derivative or immunogenic fragment thereof, may be used
with an adjuvant to provide an immunogenic composition or vaccine.
That is, the gE antigen or immunogenic derivative or immunogenic
fragment thereof may be used in a vaccination schedule in the
absence of a live attenuated strain or whole inactivated
strain.
[0091] Thus the second aspect of the invention relates to an
immunogenic composition or vaccine comprising gE or immunogenic
derivative or immunogenic fragment thereof in combination with a
TH1-adjuvant.
[0092] The invention also particularly relates to use of a
composition comprising gE or an immunogenic derivative or
immunogenic fragment thereof in combination with a TH-1 adjuvant,
in the preparation of a medicament for the prevention or
amelioration of herpes zoster reactivation and/or post herpetic
neuralgia.
[0093] The term "immunogenic derivative" in respect of gE is as
described above, along with methods to obtain such derivatives such
as fragments of gE. Immunogenic fragments as described herein are
immunogenic derivatives which retain the ability to induce an
immune response to VZV following administration to man.
[0094] In one aspect of the invention a gE truncate is used in
which gE has a C terminal truncation.
[0095] In one aspect the truncation removes from 4 to 20 percent of
the total amino acid residues at the carboxy terminal end.
[0096] In one aspect the gE is lacking the carboxy terminal anchor
region (suitably approximately amino acids 547-623 of the wild type
sequence).
[0097] In one aspect gE is a truncated gE having the sequence of
FIG. 1 herein, and as disclosed in Virus research, (Haumont et al
Vol 40, 1996 p 199-204), herein incorporated fully by
reference.
[0098] Reference to gE hereinafter includes reference to truncated
gE, or other fragments or derivative of gE, unless otherwise
apparent from the context.
[0099] In another aspect of the invention the composition comprises
full length gE.
[0100] In another aspect the gE or derivative or fragment thereof
is lyophilised. In another aspect the gE or derivative or fragment
thereof is reconstituted in a solution containing an adjuvant (such
as an adjuvant containing QS21, cholesterol and 3D MPL) before
delivery.
[0101] In one embodiment the composition or vaccine comprises gE
and a TH-1 adjuvant and does not comprise an IE63 antigen or
portion thereof. In one embodiment the composition or vaccine
comprises gE and a TH-1 adjuvant and does not comprise any other
VZV antigen. In one embodiment the composition or vaccine comprises
gE and a TH-1 adjuvant and does not comprise any other viral
antigen.
[0102] In one aspect the gE or immunogenic fragment thereof is not
in the form of a fusion protein.
[0103] In one aspect the composition or vaccine consists
essentially of QS21, a truncated VZV gE antigen and liposomes
comprising cholesterol and 3D-MPL.
[0104] In one aspect the composition or vaccine consists of 3D-MPL,
QS21, a truncated VZV gE antigen, liposomes comprising cholesterol
and a pharmaceutically acceptable carrier.
[0105] The composition may be used in the preparation of a
medicament for the prevention or amelioration of herpes zoster
reactivation and/or post herpetic neuralgia.
[0106] The composition or vaccine is suitably used in the
population of people 50 or older than 50. Suitably the population
is the population of those older than 55, 60, 65, 70, 75, 80, or
older than 80. Suitably the population is 50-70 years.
[0107] In one aspect the population of individuals are those who
have had varicella or who have had a live varicella vaccine.
[0108] Thus the invention relates to use of a composition as
described above in the preparation of a medicament for the
prevention or amelioration of herpes zoster reactivation and/or
post herpetic neuralgia in a population of people 50 or above.
[0109] The invention thus also relates to a method for the
prevention or amelioration of herpes zoster reactivation and/or
post herpetic neuralgia, the method comprising delivering to an
individual in need thereof a composition of the invention.
[0110] In one aspect the composition of the first and second
aspects of the invention are used in those individuals in whom the
varicella zoster virus has not reactivated.
[0111] The composition may be used at doses and delivery routes as
outlined above for the first aspect of the invention. Specifically
the amount of gE antigen is selected as an amount which induces an
immunoprotective response without significant, adverse side effects
in typical vaccines. Such amount will vary depending upon which
specific immunogen is employed and how it is presented. Generally,
it is expected that each dose will comprise 1-1000 .mu.g of
protein, such as 2-100 .mu.g, or 5-60 .mu.g. Where gE is used then
suitably 25-100 .mu.g gE is used, in one aspect 40-100 .mu.g of gE,
such as about 25 .mu.g, 50 .mu.g or about 100 .mu.g of gE, suitably
25 .mu.g, 50 .mu.g or 100 .mu.g gE. An optimal amount for a
particular vaccine can be ascertained by standard studies involving
observation of appropriate immune responses in subjects. Following
an initial vaccination, subjects may receive one or several booster
immunisation adequately spaced.
[0112] In one aspect the gE and adjuvant composition or vaccine is
used in a one dose delivery regime. In one aspect the gE and
adjuvant composition or vaccine is used in a two dose delivery
regime.
[0113] In one aspect the composition or vaccine of the invention is
used in a 2 dose regime with a 2 month spacing between doses.
[0114] In one aspect the TH-1 adjuvant is any adjuvant identified
above for the first aspect of the invention. In particular, a
combination of 3D MPL and QS21 may be used, for example as
disclosed in WO94/00153, or a less reactogenic composition where
the QS21 is quenched with cholesterol as disclosed in WO 96/33739
and U.S. Pat. No. 6,846,489. An alternative adjuvant comprises QS21
and 3D-MPL in an oil in water emulsion as described in WO
95/17210.
[0115] In one aspect a formulation comprises a C terminal
truncation of the VZV gE antigen, for example that given in FIG. 1,
in combination with 3D MPL and QS21.
[0116] In another aspect the invention relates to a kit comprising,
as separate components, a TH-1 adjuvant and a gE antigen or
immunogenic fragment thereof, as described above, suitable for
extemporaneous preparation of a vaccine composition. In one aspect
both components are liquids. In one aspect one component is
lyophilised and is suitable for reconstitution with the other
component. In one aspect the kit comprises a gE antigen having the
sequence of FIG. 1 and an adjuvant comprising QS21 and liposomes
comprising cholesterol and 3D MPL.
[0117] Vaccine preparation is generally described in New Trends and
Developments in Vaccines, Voller et al. (eds), University Park
Press, Baltimore, Md., 1978.
[0118] Aspects of the present invention include:
A An immunogenic composition comprising a VZV antigen or
immunogenic derivative thereof in combination with a live
attenuated VZV or whole inactivated VZV. B A method of preventing
and/or decreasing the severity of herpes zoster and/or post
herpetic neuralgia (PHN) comprising delivering to an individual an
immunogenic composition comprising a VZV antigen or immunogenic
derivative thereof in combination with a live attenuated VZV or
whole inactivated VZV. C A method of preventing and/or decreasing
the severity of herpes zoster and/or post herpetic neuralgia, the
method comprising sequential or concomitant delivery to an
individual of a VZV antigen or immunogenic derivative thereof and a
live attenuated VZV or whole inactivated VZV. D A method according
to paragraph C wherein a VZV antigen is delivered before live
attenuated VZV. E A method according to paragraph C wherein a VZV
antigen is delivered after live attenuated VZV. F A method
according to paragraph C wherein a VZV antigen is delivered
concomitantly with live attenuated VZV, preferably with each
component in a different arm of a patient. G A kit comprising a
live attenuated VZV or whole inactivated VZV and, separately, a VZV
antigen or immunogenic derivative thereof, the components suitable
for concomitant or sequential delivery, or for mixing as a single
composition prior to delivery. H A method for the manufacture of an
immunogenic composition, the method comprising combining a live
attenuated VZV or whole inactivated VZV with a VZV antigen or
immunogenic derivative thereof. I Use of a live attenuated VZV
strain in the preparation of an immunogenic composition for
preventing and/or decreasing the severity of herpes zoster and/or
post herpetic neuralgia, wherein the live attenuated VZV strain is
used in combination with a VZV antigen or immunogenic derivative
thereof. J Use of a whole inactivated VZV strain in the preparation
of an immunogenic composition for preventing and/or decreasing the
severity of herpes zoster and/or post herpetic neuralgia, wherein
the whole inactivated VZV strain is used in combination with a VZV
antigen or immunogenic derivative thereof. K Use of a VZV antigen
or immunogenic derivative thereof in the preparation of an
immunogenic composition for preventing and/or decreasing the
severity of herpes zoster and/or post herpetic neuralgia, wherein
the VZV antigen is used in combination with a live attenuated VZV
strain or whole inactivated VZV strain. L Use according to any of
paragraphs I-K wherein the antigen or derivative thereof is
delivered in a prime boost approach before the VZV strain. M Use
according to any of paragraphs I-K wherein the antigen or
derivative thereof is delivered in a prime boost approach after the
VZV strain. N Use according to any of paragraphs I-K wherein the
antigen or derivative thereof is delivered concomitantly with the
VZV strain. O Use according to any of paragraphs I-K wherein the
antigen or derivative thereof is delivered in admixture with the
VZV strain. P A use, method, kit, or composition according to any
preceding paragraph wherein the live attenuated VZV strain is the
OKA strain. Q A use method, kit, composition according to any
preceding paragraph wherein the VZV antigen the gE antigen or
immunogenic derivative thereof. R A use, method, kit, composition
or vaccine according to any preceding claim wherein the VZV antigen
is delivered with an adjuvant capable of stimulating a TH1 type
response.
[0119] The present invention is illustrated by the following, non
limiting Examples.
Example 1
[0120] Three experimental groups may be set up to study both
aspects of the invention:
TABLE-US-00001 Regime 1 50 .mu.g gE + adjuvant AS1 (MPL .RTM./QS21)
0, 2 months 2 OKA strain (Varilrix .TM.) ~10,000 pfu/dose 0, 2
months 3 Concomitant administration of 50 .mu.g gE + AS1 group 0, 2
months (as in 1) with Varilrix .TM. (as in 2) MPL .RTM. =
3D-MPL
[0121] The gE used can be truncated gE, as disclosed in FIG. 1.
[0122] Varilrix.TM. is a commercially available OKA strain.
[0123] Human volunteers (for example, 50 per group--healthy, aged
50-70 years) can be selected to be vaccinated according to the
above protocol, and results may be assessed by measuring both cell
mediated immunity and antibody responses, for example by
intracellular staining (ICS, Roederer et al. 2004 Clin. Immunol.
110: 199) or ELISA techniques respectively, these being well known
in the art.
[0124] Specific cell-mediated immunity may be evaluated by, for
example, in vitro incubation of patient PBMC with varicella-zoster
virus extracts as well as specific VZV antigens or peptides gE,
IE63 and IE62. Analysis may be made at the level of, for
example:
[0125] a Lymphoproliferation (data expressed as Stimulation Index
[SI]): GM, GM fold increase and % of responders
[0126] b Analysis of IFN.gamma. or IL2 or TNF.alpha., or CD 40L
expression by CD4 and CD8 cells by ICS (intracellular cytokine
staining): GM, GM fold increase and % of responders
[0127] Efficacy can be assessed by looking for a significant
increase in the CMI and/or antibody response in comparison with
pre-vaccination levels.
[0128] Efficacy of other antigens or approaches can be assessed
using these or similar techniques, and comparing pre-vaccination
levels with post vaccination levels.
Example 2
[0129] The experiment of Example 1 was carried out in human
volunteers of different ages, as follows:
TABLE-US-00002 Group A gE AS1 in adults 18-30 years Group B gE
delivered concomitantly with the Varilrix OKA strain 18-30 years
Group C Varilrix OKA strain alone in adults 50-70 years Group D gE
AS1 in adults 50-70 years Group E gE delivered concomitantly with
the Varilrix OKA strain 50-70 years
[0130] The vaccination schedule was as follows:
TABLE-US-00003 Group Age (Y) N Vacc 1 (Month 0) Vacc 2 (Month 2) A
18-30 10 gE-AS1 gE-AS1 B 18-30 10 gE-AS1 + Varilrix .TM. gE AS1 +
Varilrix .TM. C 50-70 45 Varilrix .TM. Varilrix .TM. D 50-70 45
gE-AS1 gE-AS1 E 50-70 45 gE-AS1 + Varilrix .TM. gE-AS1 + Varilrix
.TM.
[0131] The adjuvant AS1 comprises 3D MPL and QS21 in a quenched
form with cholesterol, and was made as described in WO9633739,
incorporated herein by reference. In particular the AS1 adjuvant
was prepared essentially as Example 1.1 of WO9633739.
[0132] The adjuvant comprises: liposomes, which in turn comprise
dioleoyl phosphatidylcholine (DOPC), cholesterol and 3D MPL [in an
amount of 1000 .mu.g DOPC, 250 .mu.g cholesterol and 50 .mu.g 3D
MPL, each value given approx per vaccine dose], QS21 [50
.mu.g/dose], PBS and water to a volume of 0.5 ml.
[0133] In the process of production of liposomes containing MPL the
DOPC (Dioleyl phosphatidylcholine), cholesterol and MPL are
dissolved in ethanol. A lipid film is formed by solvent evaporation
under vacuum. Phosphate Buffer Saline or PBS (9 mM
Na.sub.2HPO.sub.4, 41 mM KH.sub.2PO.sub.4, 100 mM NaCl) at pH 6.1
is added and the mixture is submitted to prehomogenization followed
by microfluidization at 15,000 psi (20 cycles). This leads to the
production of liposomes which are sterile filtered through a 0.22
.mu.m membrane in an aseptic (class 100) area. The sterile product
is then distributed in sterile glass containers and stored in a
cold room (+2 to +8.degree. C.). In this way the liposomes produced
contain MPL in the membrane (the "MPL in" embodiment of
WO9633739).
[0134] The truncated gE of FIG. 1 was expressed in CHO K1 cells
using standard techniques and purified using, in order, anion
exchange chromatography, hydrophobic interaction chromatography,
ion exchange chromatography, diafiltration, and nanofiltration
followed by sterilisation through a 0.22 .mu.m filter.
[0135] In particular, the following steps were used in the
purification of gE
First Stage: Anion Exchange Chromatography
[0136] The culture supernatant containing the gE (approx. 30 mg/l)
is purified, either directly after clarification of the cell
suspension or after defrosting at 4.degree. C. After transfer into
a 20-litre carboy, the pH of the supernatant is adjusted to 6.
[0137] The capture stage takes place at ambient temperature on a
chromatography column containing a Q Sepharose XL resin.
[0138] After sanitisation with sodium hydroxide, the column is
conditioned in the capture buffer (piperazine 20 mM pH 6). The
supernatant is then loaded on the column and the column is washed
with equilibration buffer and a solution of piperazine 20 mM+NaCl
150 mM pH 6.
[0139] The fraction containing the gE is then eluted with a
solution of piperazine 20 mM+NaCl 250 mM at pH 6.
Second Stage: Hydrophobic Interaction Chromatography
[0140] This chromatography stage takes place at ambient temperature
on a Toyopearl butyl-650 M resin (Tosoh Biosep).
[0141] The fraction eluted with 20 mM piperazine+250 mM NaCl in the
Q Sepharose XL stage is made up to 1 M in ammonium sulphate and
adjusted to pH 7.5.
[0142] After sanitisation with sodium hydroxide and before loading
of this fraction, the column is conditioned in the capture buffer
(50 mM KH.sub.2PO.sub.4+1 M ammonium sulphate pH 7.5). After
loading, the column is washed with buffer 50 mM
KH.sub.2PO.sub.4+100 mM (NH.sub.4).sub.2SO.sub.4 pH 7.5. The gE is
eluted with buffer 50 mM KH.sub.2PO.sub.4+25 mM
(NH.sub.4).sub.2SO.sub.4 pH 7.5.
Third Stage: Affinity Chromatography on Immobilised Metal Ion
[0143] This chromatography stage takes place at ambient temperature
on a Chelating Sepharose Fast Flow resin. This resin is saturated
in metal ion (Ni) by application of a nickel sulphate solution (1%)
and excess unbound ions (Ni), removed by washing. The gE fraction
eluted at 50 mM KH.sub.2PO.sub.4+25 mM (NH.sub.4).sub.2SO.sub.4 pH
7.5 in the hydrophobic phase is made up to 0.5 M NaCl and adjusted
to pH 7.5.
[0144] After sanitisation, the column is equilibrated in the
capture buffer (50 mM KH.sub.2PO.sub.4+0.5 M NaCl pH 7.5). The gE
solution is loaded on the column, which is then washed with a
solution of 50 mM KH.sub.2PO.sub.4+0.5 M NaCl pH 5.6. The gE is
then eluted with a buffer of 50 mM sodium acetate+0.5 M NaCl pH 5,
and neutralised with a Tris 1 M solution pH 9.5.
Fourth Stage: Diafiltration
[0145] The buffer exchange and the elimination of salts from the gE
fraction eluted at pH 5 in the previous stage are carried out by
tangential ultrafiltration. This stage is carried out entirely at
+4.degree. C.
[0146] The ultrafiltration is run by the Millipore Proflux M12
system, fitted with a 10 kDa Pellicon2 Mini membrane of regenerated
cellulose (cat: P2C010C01) of limit nominal molecular weight and a
surface area of 0.1 m.sup.2 placed in a Pellicon2 mini-cassette
housing (cat.XX42PMINI).
[0147] After rinsing with water and sanitisation with sodium
hydroxide, the whole system with membrane is rinsed with 2 litres
of modified PBS buffer (=8.1 mM Na.sub.2HPO.sub.42H.sub.2O, 1.47 mM
KH.sub.2PO.sub.4, 137 mM NaCl, 2.68 mM KCL pH 7.2) and then
equilibrated with 2 litres of the same buffer until a pH value of
7.2 is reached in the permeate.
[0148] The measurement of the permeability of the membrane is
verified. The integrity test on the membrane is carried out by
putting the system under pressure up to 1 bar before and at the end
of the diafiltration stage. If this membrane is used twice with an
interval of one week, the integrity will be tested 3 times (once
before each ultrafiltration and once after the second filtration).
The membrane is considered to be intact if the loss of pressure
recorded over 5 minutes is less than 0.1 bar. The concentration of
the gE fraction eluted at pH 5 in the affinity stage is evaluated
by measuring the optical density at 280 nm. The correlation between
the absorbance at 280 nm and the protein concentration of the gE by
microBCA is fixed at 1 OD.sub.280=1.75 mg/ml.
[0149] The solution containing the gE is dia-filtered against 10
volumes of modified PBS buffer (=8.1 mM Na.sub.2HPO.sub.42H.sub.2O,
1.47 mM KH.sub.2PO.sub.4, 137 mM NaCl, 2.68 mM KCL pH 7.2). The
pressure conditions are set such that the diafiltration period is
about 11/2-2 hours (permeate flow approx. 60 ml/min). The
diafiltration residue is then recovered and on the basis of the
baseline OD.sub.280 the membrane is rinsed with modified PBS to
give an approximate final concentration of 0.4 mg/ml.
Fifth Stage: Nanofiltration
[0150] This next stage makes it possible to eliminate viruses with
a diameter of more than 15 nm by retention. The stage is carried
out entirely at +4.degree. C.
[0151] The nanofiltration is carried out on a PLANOVA 15N filter
(mean pore size 15 nm; filtration surface area 0.12 m.sup.2 (ASAHI
cat: 15NZ-120)). Under a constant pressure of 0.45 bar, the gE
solution is filtered on the membrane and recovered on the other
side with viruses removed.
[0152] The pipes and housing (column XK50) are sanitised for 2
hours with a solution of NaOH 0.5M. The whole is then rinsed and
neutralised with the modified PBS buffer (same buffer as for the
diafiltration) until a pH value of 7.2 is achieved.
[0153] After fixing the nanofilter PLANOVA 15N (feed side) under
the casing, the filter is rinsed and equilibrated with a modified
PBS solution.
[0154] The diafiltration residue containing the gE solution is
first pre-filtered through 0.22 .mu.m (mini kleenpak OU Acropak20,
depending on the volume to be filtered) before being nano-filtered
at a constant pressure of 0.45 bar on the PLANOVA 15N.
[0155] At the end of nanofiltration, the filter is rinsed with a
sufficient volume of modified PBS to give a final concentration of
the bulk of approx. 0.3 mg/ml.
[0156] To end, the membrane is washed with 50 ml modified PBS. The
solution is recovered through the residue outlet.
[0157] The integrity tests on the PLANOVA 15N membrane are then
carried out as follows:
[0158] the first test consists of putting the membrane under
pressure (1.0 kg/cm.sup.2) and observing for formation of air
bubbles. This test detects any large splits.
[0159] the second test: elimination of particles of gold
(PARTICORPLANOVA-QCVAL4) verifies the structure of the membrane
(good distribution of large pores and capillaries).
Vaccine Composition
[0160] The gE component of the vaccine comprises 50 .mu.g gE and
the excipients sodium chloride, potassium chloride, monopotassium
phosphate, disodium phosphate and water for injection as well as
the AS1 adjuvant. The function of the inorganic salts is to ensure
isotonicity and physiological pH.
[0161] In a sterile glass container, water for injection,
concentrated phosphate buffered saline and gE antigen were mixed in
order to reach the ingredient concentration as below:
TABLE-US-00004 Quantitative Ingredients (per dose) gE 50 .mu.g
Sodium chloride (NaCl) 1603 .mu.g Disodium phosphate
(NaH.sub.2PO.sub.4) 288 .mu.g Monopotassium phosphate
(KH.sub.2PO.sub.4) 40 .mu.g Potassium chloride (KCl) 40 .mu.g Water
for injection q.s. ad 0.2 mL
[0162] The solution is mixed for 30 to 40 minutes. The pH is
checked and adjusted at 7.2+-.0.1 with HCl or NaCl or appropriate
and stir for an additional 10 minutes.
[0163] The final bulk was stored in polypropylene bottles at
-20.degree. C. and transferred to GSK Bio for filling. The vaccine
is filled into 3 ml, sterile, siliconised glass vials (0.25
ml/vial) which are closed with grey chlorobutyl rubber stoppers and
sealed with central tear-off aluminium cap. The inspected, approved
vials are then stored at -20.degree. C.
Vaccine Delivery
[0164] The gE-AS1 vaccine for administration was obtained by mixing
the liquid antigen preparation with the liquid AS1 adjuvant
immediately prior to injection (a maximum of one hour before
injection). The OKA (Varilrix.TM.) was a commercially available lot
prepared according to the manufacturer's instructions.
[0165] Vaccine formulations were as follows:
TABLE-US-00005 Vaccine gE Formulation 50 .mu.g VZV (gE) antigen in
0.2 ml volume AS1 in 0.5 ml volume Presentation Glass vial
containing liquid gE Total Dose Volume* 0.7 ml (after
reconstitution)
TABLE-US-00006 Vaccine Varilrix with diluent Formulation
Approximately 10.sup.4.0 pfu/dose 0/5 ml volume Presentation Glass
vial containing containing lyophilized vaccine for reconstitution
Total Dose Volume* 0.5 ml
[0166] The gE AS1 component was administered by intramuscular
injection.
[0167] The Varilrix component was administered by subcutaneous
injection.
Analysis of Results
[0168] The clinical trial protocol, filed in preparation for the
clinical trial, outlined the types of studies that were to be
carried out in the trial, as follows:
[0169] a Lymphoproliferation (data expressed as Stimulation Index
[SI]): GM, fold increase in GM and % of responders after
stimulation by VZV lysate.
[0170] b IFN gamma and/or IL2, TNF alpha, CD40L, CD4 and CD8
response by ICS (intracellular staining): GM,-fold increase in GM
and % of responders after stimulation by VZV lysate and gE, IE62
and IE63 peptides.
Lymphoproliferation
[0171] Peripheral blood antigen-specific lymphocytes can be
restimulated in vitro to proliferate if incubated with their
corresponding antigen. Consequently, the amount of antigen specific
lymphocytes can be estimated by counting tritiated thymidine
incorporation assay. In the present study, VZV antigen or peptide
derived from VZV proteins will be used as antigen to restimulate
VZV-specific lymphocytes. Results will be expressed as a
stimulation index (SI) which corresponds to the ratio between
antigen-specific and background lymphoproliferation.
Cytokine Flow Cytometry (CFC)
[0172] Peripheral blood antigen-specific CD4 and CD8 T cells can be
restimulated in vitro to express CD40L, IL-2, TNF alpha and IFN
gamma if incubated with their corresponding antigen. Consequently,
antigen-specific CD4 and CD8 T cells can be enumerated by flow
cytometry following conventional immunofluorescence labelling of
cellular phenotype as well as intracellular cytokines production.
In the present study, VZV antigen or peptide derived from VZV
proteins will be used as antigen to restimulate VZV-specific T
cells. Results will be expressed as a frequency of
cytokine(s)-positive CD4 or CD8 T cell within the CD4 or CD8 T cell
sub-population.
Specific Antibody (Anti-VZV and Anti-gE)
[0173] Antibody levels against VZV and gE will be measured using
classical ELISA assays.
[0174] Results of the experiment are shown in tabular form. FIGS.
2-6 present results in a graphical form for antibody (FIGS. 2-4,
see tables 1.1 a-c) and CMI responses (FIGS. 5 and 6--see table
C1/"CD4 all doubles" test with gE antigen or Varilirix, median
values).
TABLE-US-00007 HUMORAL IMMUNE RESPONSE Table I.1a Seropositivity
rates and GMTs for VZV IGG antibodies (ATP cohort for
immunogenicity) . . . Table I.1b Seropositivity rates and GMTs for
VZV.GE antibodies (ATP cohort for immunogenicity) . . . Table I.1c
Seropositivity rates and GMTs for IFA antibodies (ATP cohort for
immunogenicity) . . . Table I.2b Seroconversion rates for gE
antibody titer at each post- vaccination time point (ATP cohort for
immunogenicity) . . . Table I.3a Vaccine response for VZV antibody
titer at each post- vaccination time point (ATP cohort for
immunogenicity) . . . Table I.3b Vaccine response for gE antibody
titer at each post- vaccination time point (ATP cohort for
immunogenicity) . . . Table I.3c Vaccine response for IFA antibody
titer at each post- vaccination time point (ATP cohort for
immunogenicity) . . .
TABLE-US-00008 TABLE I.1a Seropositivity rates and GMTs for VZV IGG
antibodies (ATP cohort for immunogenicity) >=50 MIU/ML GMT 95%
CI 95% CI Antibody Group Timing N n % LL UL value LL UL Min Max VZV
IGG gE/Y PRE 10 10 100 69.2 100 1875.9 1077.2 3266.8 455.0 4634.0
PI(M1) 10 10 100 69.2 100 14843.0 8457.7 26049.0 6051.0 65242.0
PI(M2) 10 10 100 69.2 100 10697.6 6768.2 16908.4 3167.0 29622.0
PII(M3) 10 10 100 69.2 100 14330.6 10492.9 19571.9 7173.0 30894.0
gEVAR/Y PRE 10 10 100 69.2 100 1047.4 519.3 2112.6 300.0 5754.0
PI(M1) 10 10 100 69.2 100 12859.4 7063.0 23412.8 3346.0 49163.0
PI(M2) 10 10 100 69.2 100 10072.2 5631.4 18014.7 3678.0 36289.0
PII(M3) 10 10 100 69.2 100 15245.7 9930.9 23404.8 6381.0 36534.0
VAR/E PRE 45 45 100 92.1 100 856.9 647.5 1134.1 100.0 5377.0 PI(M1)
45 45 100 92.1 100 2538.8 2072.3 3110.3 288.0 8034.0 PI(M2) 45 45
100 92.1 100 2292.2 1880.6 2793.9 374.0 7549.0 PII(M3) 45 45 100
92.1 100 2338.3 1933.6 2827.7 523.0 16994.0 gE/E PRE 45 45 100 92.1
100 940.1 744.2 1187.6 208.0 4221.0 PI(M1) 45 45 100 92.1 100
5897.4 4594.7 7569.5 659.0 27042.0 PI(M2) 45 45 100 92.1 100 4523.0
3570.6 5729.4 598.0 19268.0 PII(M3) 45 45 100 92.1 100 9083.9
7437.6 11094.5 2493.0 42073.0 gEVAR/E PRE 44 44 100 92.0 100 1165.1
954.6 1422.1 209.0 5558.0 PI(M1) 44 44 100 92.0 100 8371.7 6637.2
10559.5 2509.0 56066.0 PI(M2) 44 44 100 92.0 100 6849.1 5422.6
8650.8 1753.0 55958.0 PII(M3) 44 44 100 92.0 100 9849.1 8201.7
11827.3 3528.0 38664.0 gE/Y = gE-AS1/18-30 years gEVAR/Y = gE-AS1 +
Varilrix/18-30 years VAR/E = Varilrix/50-70 years gE/E =
gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70 years GMC =
geometric mean antibody concentration calculated on all subjects N
= number of subjects with available results n/% = number/percentage
of subjects with concentration within the specified range 95% CI =
95% confidence interval; LL = Lower Limit, UL = Upper Limit MIN/MAX
= Minimum/Maximum PRE = Pre-vaccination dose 1 PI(M1) =
Post-vacciantion dose 1 (Month 1) PI(M2) = Post-vaccination dose 1
(Month 2) PII(M3) = Post-vaccination dose 2 (Month 3)
TABLE-US-00009 TABLE I.1b Seropositivity rates and GMTs for VZV.GE
antibodies (ATP cohort for immunogenicity) >=109 ELU/ML GMT 95%
CI 95% CI Antibody Group Timing N n % LL UL value LL UL Min Max
VZV.GE gE/Y PRE 10 8 80.0 44.4 97.5 302.6 120.5 759.9 <109.0
2169.0 PI(M1) 10 10 100 69.2 100 18365.0 9610.6 35094.1 5697.0
106829.0 PI(M2) 10 10 100 69.2 100 11076.6 7037.4 17433.9 3528.0
30190.0 PII(M3) 10 10 100 69.2 100 15842.6 11543.4 21743.1 7502.0
30487.0 gEVAR/Y PRE 10 7 70.0 34.8 93.3 190.3 96.4 375.6 <109.0
661.0 PI(M1) 10 10 100 69.2 100 16225.7 8657.3 30410.6 3613.0
58950.0 PI(M2) 10 10 100 69.2 100 11554.7 6312.5 21150.4 3656.0
47423.0 PII(M3) 10 10 100 69.2 100 18101.2 11384.7 28780.0 7649.0
44539.0 VAR/E PRE 44 35 79.5 64.7 90.2 266.9 189.6 375.8 <109.0
5866.0 PI(M1) 45 45 100 92.1 100 1011.3 770.0 1328.2 177.0 6386.0
PI(M2) 45 45 100 92.1 100 948.1 701.6 1281.2 127.0 6759.0 PII(M3)
45 45 100 92.1 100 1146.9 841.5 1563.0 164.0 16249.0 gE/E PRE 45 37
82.2 67.9 92.0 231.1 178.8 298.7 <109.0 899.0 PI(M1) 45 45 100
92.1 100 6099.1 4401.9 8450.8 367.0 40101.0 PI(M2) 45 45 100 92.1
100 4844.2 3406.5 6888.8 288.0 42488.0 PII(M3) 45 45 100 92.1 100
14816.8 12122.2 18110.2 3047.0 58792.0 gEVAR/E PRE 44 42 95.5 84.5
99.4 336.1 268.0 421.5 <109.0 1531.0 PI(M1) 44 44 100 92.0 100
8272.6 6071.1 11272.4 363.0 54878.0 PI(M2) 44 44 100 92.0 100
7870.4 5937.0 10433.4 1512.0 84465.0 PII(M3) 44 44 100 92.0 100
16616.0 13972.3 19760.0 4774.0 61558.0 gE/Y = gE-AS1/18-30 years
gEVAR/Y = gE-AS1 + Varilrix/18-30 years VAR/E = Varilrix/50-70
years gE/E = gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70
years GMC = geometric mean antibody concentration calculated on all
subjects N = number of subjects with available results n/% =
number/percentage of subjects with concentration within the
specified range 95% CI = 95% confidence interval; LL = Lower Limit,
UL = Upper Limit MIN/MAX = Minimum/Maximum PRE = Pre-vaccination
dose 1 PI(M1) = Post-vacciantion dose 1 (Month 1) PI(M2) =
Post-vaccination dose 1 (Month 2) PII(M3) = Post-vaccination dose 2
(Month 3)
TABLE-US-00010 TABLE I.1c Seropositivity rates and GMTs for IFA
antibodies (ATP cohort for immunogenicity) >=4 1/DIL GMT 95% CI
95% CI Antibody Group Timing N n % LL UL value LL UL Min Max IFA
gE/Y PRE 10 10 100 69.2 100 1351.2 691.9 2638.8 256.0 4096.0 PI(M1)
10 10 100 69.2 100 10809.4 6040.0 19345.1 4096.0 65536.0 PI(M2) 10
10 100 69.2 100 12416.8 6631.6 23248.7 2048.0 32768.0 PII(M3) 10 10
100 69.2 100 14263.1 10423.6 19516.8 8192.0 32768.0 gEVAR/Y PRE 10
10 100 69.2 100 776.0 321.6 1872.5 256.0 16384.0 PI(M1) 10 10 100
69.2 100 9410.1 5638.8 15703.7 4096.0 32768.0 PI(M2) 10 10 100 69.2
100 14263.1 8546.9 23802.4 8192.0 65536.0 PII(M3) 10 10 100 69.2
100 12416.8 8173.6 18862.6 8192.0 32768.0 VAR/E PRE 45 45 100 92.1
100 686.1 508.5 925.6 128.0 8192.0 PI(M1) 45 45 100 92.1 100 2702.4
2115.6 3451.9 512.0 32768.0 PI(M2) 45 45 100 92.1 100 1838.7 1454.0
2325.2 256.0 16384.0 PII(M3) 45 45 100 92.1 100 2144.9 1707.4
2694.4 256.0 8192.0 gE/E PRE 45 45 100 92.1 100 597.3 452.8 787.8
128.0 8192.0 PI(M1) 45 45 100 92.1 100 6402.6 4799.2 8541.8 512.0
32768.0 PI(M2) 45 45 100 92.1 100 4356.3 3247.0 5844.7 256.0
32768.0 PII(M3) 45 45 100 92.1 100 10163.5 8426.4 12258.7 1024.0
32768.0 gEVAR/E PRE 44 44 100 92.0 100 783.4 620.8 988.7 128.0
4096.0 PI(M1) 44 44 100 92.0 100 9004.1 6946.4 11671.3 2048.0
65536.0 PI(M2) 44 44 100 92.0 100 6169.4 4908.2 7754.6 2048.0
32768.0 PII(M3) 44 44 100 92.0 100 11225.9 9284.5 13573.3 4096.0
32768.0 gE/Y = gE-AS1/18-30 years gEVAR/Y = gE-AS1 + Varilrix/18-30
years VAR/E = Varilrix/50-70 years gE/E = gE-AS1/50-70 years
gEVAR/E = gE-AS1 + Varilrix/50-70 years GMT = geometric mean
antibody titre calculated on all subjects N = number of subjects
with available results n/% = number/percentage of subjects with
titre within the specified range 95% CI = 95% confidence interval;
LL = Lower Limit, UL = Upper Limit MIN/MAX = Minimum/Maximum PRE =
Pre-vaccination dose 1 PI(M1) = Post-vacciantion dose 1 (Month 1)
PI(M2) = Post-vaccination dose 1 (Month 2) PII(M3) =
Post-vaccination dose 2 (Month 3)
TABLE-US-00011 TABLE I.2b Seroconversion rates for gE antibody
titer at each post- vaccination time point (ATP cohort for
immunogenicity) Seroconversion 95% CI Group Timing N n % LL UL gE/Y
Month 1 2 2 100.0 15.8 100.0 Month 2 2 2 100.0 15.8 100.0 Month 3 2
2 100.0 15.8 100.0 gEVAR/Y Month 1 3 3 100.0 29.2 100.0 Month 2 3 3
100.0 29.2 100.0 Month 3 3 3 100.0 29.2 100.0 VAR/E Month 1 9 9
100.0 66.4 100.0 Month 2 9 9 100.0 66.4 100.0 Month 3 9 9 100.0
66.4 100.0 gE/E Month 1 8 8 100.0 63.1 100.0 Month 2 8 8 100.0 63.1
100.0 Month 3 8 8 100.0 63.1 100.0 gEVAR/E Month 1 2 2 100.0 15.8
100.0 Month 2 2 2 100.0 15.8 100.0 Month 3 2 2 100.0 15.8 100.0
gE/Y = gE-AS1/18-30 years gEVAR/Y = gE-AS1 + Varilrix/18-30 years
VAR/E = Varilrix/50-70 years gE/E = gE-AS1/50-70 years gEVAR/E =
gE-AS1 + Varilrix/50-70 years N = number of seronegative subjects
at day 0 n/% = number/percentage of initially seronegative subjects
who became seropositive at the specified post-vaccination time
point 95% CI = 95% confidence interval; LL = Lower Limit, UL =
Upper Limit
TABLE-US-00012 TABLE I.3a Vaccine response for VZV antibody titer
at each post- vaccination time point (ATP cohort for
immunogenicity) Vaccine response 95% CI Group Timing N n % LL UL
gE/Y Month 1 10 7 70.0 34.8 93.3 Month 2 10 6 60.0 26.2 87.8 Month
3 10 9 90.0 55.5 99.7 gEVAR/Y Month 1 10 10 100.0 69.2 100.0 Month
2 10 10 100.0 69.2 100.0 Month 3 10 10 100.0 69.2 100.0 VAR/E Month
1 45 11 24.4 12.9 39.5 Month 2 45 10 22.2 11.2 37.1 Month 3 45 11
24.4 12.9 39.5 gE/E Month 1 45 29 64.4 48.8 78.1 Month 2 45 24 53.3
37.9 68.3 Month 3 45 39 86.7 73.2 94.9 gEVAR/E Month 1 44 33 75.0
59.7 86.8 Month 2 44 27 61.4 45.5 75.6 Month 3 44 38 86.4 72.6 94.8
gE/Y = gE-AS1/18-30 years gEVAR/Y = gE-AS1 + Varilrix/18-30 years
VAR/E = Varilrix/50-70 years gE/E = gE-AS1/50-70 years gEVAR/E =
gE-AS1 + Varilrix/50-70 years N = number of seropositive subjects
at day 0 n/% = number/percentage of initially seropositive subjects
with a four-fold increase at the specified post-vaccination time
point 95% CI = 95% confidence interval; LL = Lower Limit, UL =
Upper Limit
[0175] Analysis of CMI responses is given below
TABLE-US-00013 LIST OF TABLES Table C.1 Intracellular Cytokine
Staining (ICS): Descriptive Statistics on CD4 T cells at each time
point (Total vaccinated Cohort) . . . Supplementary Table C.1
Intracellular Cytokine Staining (ICS): Descriptive Statistics on
CD8 T cells at each time point (Total vaccinated Cohort) . . .
Table C.2 Intracellular Cytokine Staining (ICS): Inferential
statistics: P-values from Kruskal-Wallis Tests for CD4 T cells at
each time point (Total Vaccinated Cohort) . . . Supplementary Table
C.2 Intracellular Cytokine Staining (ICS): Inferential statistics:
P- values from Kruskal-Wallis Tests for CD8 T cells at each time
point (Total Vaccinated Cohort) . . . Table C.3 Intracellular
Cytokine Staining (ICS): Descriptive Statistics on CD4 T cells at
POST-PRE (Total vaccinated Cohort) . . . Supplementary Table C.3
Intracellular Cytokine Staining (ICS): Descriptive Statistics on
CD8 T cells at POST-PRE (Total vaccinated Cohort) . . . Table C.4
Intracellular Cytokine Staining (ICS): Inferential statistics:
P-values from Kruskal-Wallis Tests for CD4 T cells at POST-PRE
(Total Vaccinated Cohort) . . . Supplementary Table C.4
Intracellular Cytokine Staining (ICS): Inferential statistics: P-
values from Kruskal-Wallis Tests for CD8 T cells at POST-PRE (Total
Vaccinated Cohort) . . .
TABLE-US-00014 TABLE C.1 Intracellular Cytokine Staining (ICS):
Descriptive Statistics on CD4 T cells at each time point (Total
vaccinated Cohort) Test Antigen Group Timing N N miss. Mean SD Min
Q1 Median Q3 Max CD4-ALL Pool gE gE/Y Day 0 9 1 213.44 202.59 1.00
1.00 139.00 385.00 490.00 DOUBLES Month 1 9 1 1383.78 1629.94
256.00 342.00 807.00 1333.00 5207.00 Month 2 9 1 1787.56 1818.51
497.00 677.00 919.00 1775.00 5539.00 Month 3 9 1 2739.89 1856.98
581.00 1770.00 2234.00 2909.00 6963.00 gEVAR/Y Day 0 10 0 253.20
246.76 1.00 1.00 246.00 391.00 783.00 Month 1 10 0 1179.70 991.68
1.00 567.00 979.00 1364.00 3535.00 Month 2 9 1 1546.44 886.57
116.00 846.00 1996.00 2092.00 2538.00 Month 3 10 0 3298.50 1477.17
1699.00 1970.00 2944.00 4924.00 5840.00 VAR/E Day 0 44 1 299.98
922.48 1.00 1.00 126.00 238.00 6152.00 Month 1 43 2 458.28 1256.04
1.00 89.00 194.00 294.00 8000.00 Month 2 44 1 246.09 243.71 1.00
87.50 177.00 339.00 1252.00 Month 3 45 0 476.40 1149.12 1.00 1.00
151.00 365.00 6264.00 gE/E Day 0 44 1 166.27 180.38 1.00 1.00
104.50 291.50 657.00 Month 1 42 3 849.10 1090.91 1.00 226.00 540.00
949.00 4487.00 Month 2 43 2 522.12 577.93 1.00 115.00 402.00 627.00
2372.00 Month 3 43 2 3221.91 2534.70 1.00 1184.00 2323.00 4767.00
11480.00 gEVAR/E Day 0 45 0 206.02 265.71 1.00 36.00 158.00 249.00
1552.00 Month 1 44 1 826.70 614.40 1.00 322.50 770.00 1158.50
2927.00 Month 2 45 0 509.16 411.89 1.00 166.00 447.00 737.00
1553.00 Month 3 45 0 2918.04 2522.39 8.00 1081.00 1902.00 4251.00
10468.00 Varilrix gE/Y Day 0 9 1 1045.78 770.48 369.00 544.00
761.00 1067.00 2590.00 Month 1 9 1 1302.67 1378.05 1.00 314.00
597.00 1877.00 4109.00 Month 2 9 1 1656.00 1224.33 561.00 890.00
1238.00 2019.00 4494.00 Month 3 9 1 1816.33 994.51 590.00 1241.00
1368.00 2540.00 3755.00 gEVAR/Y Day 0 10 0 1188.40 580.19 412.00
783.00 1058.50 1535.00 2332.00 Month 1 10 0 1090.00 1168.31 1.00
610.00 770.00 1123.00 4267.00 Month 2 9 1 2659.78 1316.29 873.00
1868.00 2282.00 4171.00 4246.00 Month 3 10 0 3369.70 2127.99
1147.00 1738.00 2854.50 4494.00 7745.00 VAR/E Day 0 44 1 581.02
635.92 1.00 129.50 415.00 753.50 3327.00 Month 1 43 2 992.35
1093.91 1.00 268.00 661.00 1447.00 5359.00 Month 2 44 1 815.75
928.07 1.00 222.50 517.00 1065.00 4575.00 Month 3 45 0 984.67
832.16 1.00 364.00 774.00 1278.00 3287.00 gE/E Day 0 44 1 758.18
982.52 1.00 81.50 395.00 929.50 4479.00 Month 1 42 3 1216.60
1674.07 1.00 307.00 591.00 1410.00 7779.00 Month 2 43 2 869.35
1017.90 1.00 217.00 543.00 1072.00 4556.00 Month 3 43 2 2192.42
1977.14 56.00 840.00 1862.00 2557.00 9167.00 gEVAR/E Day 0 45 0
510.73 512.13 1.00 219.00 376.00 675.00 2218.00 Month 1 44 1
1179.50 1005.41 1.00 434.50 982.00 1529.00 4478.00 Month 2 45 0
961.78 915.99 1.00 292.00 704.00 1078.00 3975.00 Month 3 45 0
2484.47 1713.46 109.00 1270.00 2008.00 3429.00 6585.00 CD4- Pool gE
gE/Y Day 0 9 1 204.67 193.66 1.00 1.00 139.00 356.00 490.00 CD40L
Month 1 9 1 1347.33 1570.63 244.00 400.00 768.00 1273.00 5021.00
Month 2 9 1 1724.44 1737.50 466.00 660.00 869.00 1859.00 5252.00
Month 3 9 1 2567.89 1723.12 514.00 1744.00 1905.00 2813.00 6414.00
gEVAR/Y Day 0 10 0 253.80 240.23 1.00 41.00 265.50 361.00 752.00
Month 1 10 0 1122.30 999.38 1.00 480.00 948.00 1266.00 3534.00
Month 2 9 1 1520.33 898.50 116.00 732.00 2045.00 2068.00 2633.00
Month 3 10 0 3169.30 1452.28 1555.00 1857.00 2910.50 4539.00
5840.00 VAR/E Day 0 44 1 292.50 872.61 1.00 1.00 119.50 238.50
5812.00 Month 1 43 2 444.93 1175.20 1.00 70.00 214.00 316.00
7453.00 Month 2 44 1 242.66 242.66 1.00 95.00 178.00 333.00 1252.00
Month 3 45 0 465.67 1124.33 1.00 18.00 154.00 365.00 6072.00 gE/E
Day 0 44 1 158.55 173.14 1.00 5.50 98.50 290.50 564.00 Month 1 42 3
840.79 1061.43 1.00 237.00 530.00 910.00 4428.00 Month 2 43 2
529.58 558.23 1.00 147.00 365.00 645.00 2251.00 Month 3 43 2
3133.65 2453.67 1.00 1172.00 2294.00 4703.00 10561.00 gEVAR/E Day 0
45 0 206.16 257.80 1.00 35.00 167.00 278.00 1525.00 Month 1 44 1
814.57 606.93 1.00 314.00 773.00 1153.00 2762.00 Month 2 45 0
515.11 400.69 1.00 204.00 459.00 737.00 1523.00 Month 3 45 0
2898.40 2512.50 97.00 1073.00 1915.00 4200.00 10418.00 Varilrix
gE/Y Day 0 9 1 1027.78 746.06 373.00 544.00 761.00 1033.00 2507.00
Month 1 9 1 1264.11 1345.95 1.00 277.00 554.00 1854.00 3992.00
Month 2 9 1 1609.67 1112.38 561.00 978.00 1238.00 1992.00 4094.00
Month 3 9 1 1750.67 987.16 573.00 1039.00 1329.00 2556.00 3598.00
gEVAR/Y Day 0 10 0 1174.60 581.33 379.00 747.00 1061.50 1535.00
2327.00 Month 1 10 0 1051.70 1159.05 1.00 610.00 706.50 1019.00
4211.00 Month 2 9 1 2623.56 1344.14 873.00 1726.00 2281.00 4168.00
4273.00 Month 3 10 0 3229.20 2099.64 1089.00 1624.00 2797.00
4400.00 7592.00 VAR/E Day 0 44 1 574.57 617.51 1.00 129.50 410.00
785.50 3268.00 Month 1 43 2 976.98 1068.41 1.00 282.00 661.00
1386.00 5305.00 Month 2 44 1 801.14 922.73 1.00 230.50 519.50
1044.00 4601.00 Month 3 45 0 965.73 817.64 1.00 409.00 733.00
1255.00 3264.00 gE/E Day 0 44 1 742.41 978.83 1.00 85.00 367.50
906.00 4506.00 Month 1 42 3 1184.31 1577.03 1.00 307.00 608.00
1410.00 7748.00 Month 2 43 2 868.70 1000.52 2.00 195.00 521.00
1020.00 4435.00 Month 3 43 2 2143.84 1963.92 64.00 840.00 1849.00
2508.00 9008.00 gEVAR/E Day 0 45 0 506.91 508.06 1.00 177.00 367.00
586.00 2191.00 Month 1 44 1 1172.11 1005.52 19.00 452.00 949.50
1507.50 4478.00 Month 2 45 0 963.98 907.43 7.00 325.00 680.00
1091.00 3975.00 Month 3 45 0 2438.44 1675.69 73.00 1239.00 2008.00
3372.00 6538.00 CD4-IFN.gamma. Pool gE gE/Y Day 0 9 1 111.00 101.80
1.00 1.00 104.00 154.00 295.00 Month 1 9 1 966.89 1288.87 1.00
212.00 499.00 824.00 3972.00 Month 2 9 1 1263.67 1422.73 193.00
498.00 665.00 1298.00 4705.00 Month 3 9 1 1952.44 1798.75 358.00
1189.00 1302.00 2103.00 6292.00 gEVAR/Y Day 0 10 0 177.30 118.58
24.00 112.00 138.00 242.00 437.00 Month 1 10 0 823.50 948.60 1.00
272.00 591.00 851.00 3334.00 Month 2 9 1 1087.22 751.60 39.00
503.00 1290.00 1626.00 2226.00 Month 3 10 0 2285.60 1272.75 1146.00
1243.00 1967.00 2691.00 5437.00 VAR/E Day 0 44 1 174.95 478.34 1.00
29.50 54.50 154.00 3179.00 Month 1 43 2 290.14 764.08 1.00 47.00
92.00 220.00 4988.00 Month 2 44 1 172.70 201.53 1.00 40.50 117.00
216.00 1105.00 Month 3 45 0 279.49 586.44 1.00 1.00 100.00 193.00
3226.00 gE/E Day 0 44 1 128.91 150.94 1.00 10.50 75.50 197.00
586.00 Month 1 42 3 513.38 768.05 1.00 55.00 250.00 520.00 3471.00
Month 2 43 2 293.86 414.90 1.00 48.00 144.00 333.00 1894.00 Month 3
43 2 1672.33 1602.24 1.00 596.00 1307.00 2104.00 6309.00 gEVAR/E
Day 0 45 0 123.78 173.17 1.00 36.00 76.00 159.00 1078.00 Month 1 44
1 474.86 405.90 1.00 161.00 326.00 746.50 1536.00 Month 2 45 0
295.87 316.87 1.00 68.00 190.00 425.00 1132.00 Month 3 45 0 1516.18
1303.21 67.00 620.00 1024.00 2188.00 5829.00 Varilrix gE/Y Day 0 9
1 855.44 722.82 310.00 391.00 577.00 839.00 2466.00 Month 1 9 1
1051.89 1228.06 1.00 247.00 384.00 1462.00 3637.00 Month 2 9 1
1283.67 1065.12 410.00 475.00 966.00 1482.00 3808.00 Month 3 9 1
1362.44 910.53 448.00 833.00 1045.00 1873.00 3297.00 gEVAR/Y Day 0
10 0 946.70 536.60 438.00 548.00 822.50 1097.00 2162.00 Month 1 10
0 921.30 1143.99 1.00 365.00 586.00 1047.00 4042.00 Month 2 9 1
2176.56 1249.74 838.00 1181.00 1711.00 3439.00 3885.00 Month 3 10 0
2715.60 1892.41 716.00 1327.00 2380.50 3836.00 6992.00 VAR/E Day 0
44 1 481.73 550.51 1.00 95.50 367.50 600.00 2960.00 Month 1 43 2
827.30 941.73 1.00 253.00 529.00 1287.00 4765.00 Month 2 44 1
641.86 757.75 3.00 160.00 444.50 797.50 3922.00 Month 3 45 0 772.67
690.72 1.00 244.00 580.00 1003.00 2797.00 gE/E Day 0 44 1 629.98
847.28 1.00 72.50 228.00 819.00 3920.00 Month 1 42 3 972.52 1380.48
1.00 186.00 419.00 1154.00 6695.00 Month 2 43 2 673.05 857.44 1.00
137.00 371.00 730.00 4126.00 Month 3 43 2 1581.98 1625.99 58.00
618.00 1381.00 1735.00 7796.00 gEVAR/E Day 0 45 0 386.82 395.99
1.00 92.00 263.00 483.00 1579.00 Month 1 44 1 937.20 879.65 1.00
338.50 656.50 1132.50 3637.00 Month 2 45 0 786.73 775.90 1.00
343.00 566.00 901.00 3205.00 Month 3 45 0 1769.78 1236.32 1.00
884.00 1439.00 2289.00 4992.00 CD4-IL2 Pool gE gE/Y Day 0 9 1
166.44 178.37 1.00 1.00 66.00 337.00 420.00 Month 1 9 1 1259.44
1464.99 240.00 371.00 768.00 1119.00 4701.00 Month 2 9 1 1662.11
1698.78 402.00 593.00 892.00 1721.00 5076.00 Month 3 9 1 2317.67
1497.67 410.00 1423.00 2011.00 2595.00 5534.00 gEVAR/Y Day 0 10 0
237.20 233.77 1.00 50.00 231.00 349.00 751.00 Month 1 10 0 987.90
796.31 1.00 509.00 751.50 1361.00 2799.00 Month 2 9 1 1404.67
723.26 270.00 802.00 1905.00 1975.00 2029.00 Month 3 10 0 2747.80
1210.11 1436.00 1563.00 2367.50 4296.00 4451.00 VAR/E Day 0 44 1
269.98 833.31 1.00 33.00 119.00 212.00 5582.00 Month 1 43 2 388.26
995.89 1.00 58.00 164.00 293.00 6248.00 Month 2 44 1 211.18 231.64
1.00 53.50 158.00 286.50 1182.00 Month 3 45 0 397.51 916.41 1.00
2.00 146.00 329.00 4728.00 gE/E Day 0 44 1 149.86 153.06 1.00 1.00
95.50 252.00 550.00 Month 1 42 3 761.98 992.38 5.00 157.00 451.50
800.00 4039.00 Month 2 43 2 467.58 523.51 1.00 103.00 329.00 541.00
2094.00 Month 3 43 2 2809.07 2307.31 1.00 1037.00 2177.00 4347.00
10316.00 gEVAR/E Day 0 45 0 165.16 246.51 1.00 35.00 116.00 203.00
1551.00 Month 1 44 1 712.59 540.93 1.00 228.00 728.50 1048.00
2381.00 Month 2 45 0 465.58 354.74 1.00 184.00 385.00 667.00
1239.00 Month 3 45 0 2550.27 2304.36 1.00 945.00 1713.00 3910.00
9561.00 Varilrix gE/Y Day 0 9 1 941.44 645.88 446.00 492.00 617.00
967.00 2217.00 Month 1 9 1 1092.22 1164.69 1.00 249.00 469.00
1576.00 3480.00 Month 2 9 1 1482.00 1067.28 393.00 906.00 1148.00
1715.00 3938.00 Month 3 9 1 1551.22 851.75 484.00 869.00 1275.00
2083.00 3112.00 gEVAR/Y Day 0 10 0 1074.70 505.73 414.00 748.00
988.00 1265.00 2094.00 Month 1 10 0 903.00 887.48 1.00 486.00
650.50 880.00 3256.00 Month 2 9 1 2403.33 1122.05 989.00 1679.00
2086.00 3309.00 4127.00 Month 3 10 0 2791.90 1716.41 1043.00
1472.00 2186.50 3978.00 6004.00 VAR/E Day 0 44 1 536.80 592.24 1.00
135.50 403.50 710.00 3190.00 Month 1 43 2 866.63 984.53 1.00 220.00
608.00 1154.00 4813.00 Month 2 44 1 711.09 830.59 1.00 204.50
452.50 907.50 4261.00 Month 3 45 0 828.55 733.60 1.00 331.00 632.00
1076.00 2948.00 gE/E Day 0 44 1 701.02 866.81 1.00 97.50 280.00
885.50 3493.00 Month 1 42 3 1076.55 1440.99 1.00 233.00 548.00
1316.00 6726.00 Month 2 43 2 785.88 884.51 1.00 167.00 515.00
971.00 3707.00 Month 3 43 2 1866.37 1721.01 1.00 670.00 1569.00
2295.00 7835.00 gEVAR/E Day 0 45 0 465.67 489.70 1.00 142.00 334.00
557.00 2107.00 Month 1 44 1 1056.59 942.76 18.00 441.50 802.00
1408.50 4071.00 Month 2 45 0 885.89 853.16 1.00 328.00 628.00
1013.00 3815.00 Month 3 45 0 2114.20 1524.74 72.00 993.00 1660.00
3199.00 5671.00 CD4-TNF.alpha. Pool gE gE/Y Day 0 9 1 99.33 94.45
1.00 32.00 68.00 189.00 245.00 Month 1 9 1 659.56 926.82 114.00
155.00 201.00 757.00 2960.00 Month 2 9 1 891.11 1360.20 71.00
261.00 347.00 868.00 4435.00 Month 3 9 1 1423.33 1570.82 238.00
494.00 914.00 1350.00 5269.00 gEVAR/Y Day 0 10 0 117.00 157.25 1.00
1.00 28.00 265.00 380.00 Month 1 10 0 649.30 523.37 34.00 352.00
551.00 805.00 1952.00 Month 2 9 1 804.67 520.96 39.00 504.00 717.00
1191.00 1578.00 Month 3 10 0 1770.00 890.04 470.00 1223.00 1865.00
2258.00 3054.00 VAR/E Day 0 44 1 211.11 697.05 1.00 1.00 73.00
158.00 4652.00 Month 1 43 2 271.98 755.07 1.00 43.00 109.00 217.00
4812.00 Month 2 44 1 149.80 172.34 1.00 38.50 109.00 233.00 1007.00
Month 3 45 0 291.98 713.57 1.00 1.00 108.00 198.00 4213.00 gE/E Day
0 44 1 125.23 143.92 1.00 1.00 54.00 213.00 531.00 Month 1 42 3
444.05 585.76 1.00 64.00 283.50 473.00 2574.00 Month 2 43 2 319.30
371.77 1.00 103.00 225.00 425.00 1808.00 Month 3 43 2 1902.56
1602.67 1.00 779.00 1414.00 2860.00 7655.00 gEVAR/E Day 0 45 0
153.76 193.78 1.00 33.00 129.00 190.00 1025.00 Month 1 44 1 432.39
326.71 1.00 135.50 410.50 694.50 1479.00 Month 2 45 0 318.69 273.85
1.00 96.00 265.00 438.00 1097.00 Month 3 45 0 1662.40 1570.03 1.00
671.00 1091.00 2194.00 6609.00 Varilrix gE/Y Day 0 9 1 754.44
694.51 260.00 286.00 315.00 902.00 2217.00 Month 1 9 1 812.11
925.84 1.00 131.00 370.00 1275.00 2851.00 Month 2 9 1 1204.56
980.05 420.00 690.00 738.00 1436.00 3581.00 Month 3 9 1 1129.33
872.35 286.00 592.00 719.00 1539.00 2972.00 gEVAR/Y Day 0 10 0
816.50 449.18 239.00 597.00 733.00 1024.00 1577.00 Month 1 10 0
660.20 520.39 217.00 342.00 482.50 895.00 1965.00 Month 2 9 1
1789.67 952.77 418.00 1084.00 1396.00 2627.00 3273.00 Month 3 10 0
2016.50 1220.42 437.00 1321.00 1774.00 3179.00 4156.00 VAR/E Day 0
44 1 467.02 590.33 1.00 86.50 303.00 607.50 3075.00 Month 1 43 2
751.72 934.46 1.00 223.00 405.00 1079.00 4794.00 Month 2 44 1
638.30 813.04 1.00 215.50 340.00 812.00 3974.00 Month 3 45 0 711.04
662.11 1.00 268.00 535.00 865.00 2893.00 gE/E Day 0 44 1 623.09
800.32 1.00 96.50 257.50 833.50 3426.0 Month 1 42 3 862.48 1203.33
1.00 197.00 389.00 1091.00 5641.0 Month 2 43 2 664.63 820.99 22.00
142.00 334.00 845.00 3582.0 Month 3 43 2 1508.67 1419.81 1.00
537.00 1353.00 1836.00 6451.0 gEVAR/E Day 0 45 0 405.42 379.35 1.00
168.00 260.00 558.00 1616.0 Month 1 44 1 814.70 780.82 1.00 274.50
574.50 989.00 3311.0 Month 2 45 0 699.20 628.49 1.00 232.00 549.00
898.00 2501.0 Month 3 45 0 1634.80 1160.30 73.00 854.00 1450.00
2097.00 4405.0 gE/Y = gE-AS1/18-30 years gEVAR/Y = gE-AS1 +
Varilrix/18-30 years VAR/E = Varilrix/50-70 years gE/E =
gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70 years N =
number of subjects with available results N miss. = number of
subjects with missing results SD = Standard Deviation Min, Max =
Minimum, Maximum Q1, Q3 = First, Third quartile
TABLE-US-00015 SUPPLEMENTARY TABLE C.1 Intracellular Cytokine
Staining (ICS): Descriptive Statistics on CD8 T cells at each time
point (Total vaccinated Cohort) Test Antigen Group Timing N N miss.
Mean SD Min Q1 Median Q3 Max CD8-ALL Pool gE gE/Y Day 0 9 1 37.78
48.76 1.00 1.00 1.00 68.00 137.00 DOUBLES Month 1 9 1 61.78 111.15
1.00 1.00 1.00 68.00 345.00 Month 2 9 1 587.67 1585.68 1.00 1.00
1.00 137.00 4811.00 Month 3 9 1 38.67 50.04 1.00 1.00 1.00 67.00
141.00 gEVAR/Y Day 0 10 0 151.00 246.45 1.00 1.00 35.00 206.00
742.00 Month 1 10 0 34.30 65.70 1.00 1.00 1.00 64.00 205.00 Month 2
9 1 39.00 114.00 1.00 1.00 1.00 1.00 343.00 Month 3 10 0 177.80
313.49 1.00 1.00 36.00 272.00 1013.00 VAR/E Day 0 44 1 40.32 79.95
1.00 1.00 1.00 68.00 348.00 Month 1 43 2 33.16 59.81 1.00 1.00 1.00
72.00 216.00 Month 2 43 2 41.14 75.20 1.00 1.00 1.00 70.00 284.00
Month 3 45 0 29.20 62.99 1.00 1.00 1.00 1.00 286.00 gE/E Day 0 44 1
23.64 50.66 1.00 1.00 1.00 1.00 221.00 Month 1 42 3 35.17 82.74
1.00 1.00 1.00 28.00 422.00 Month 2 43 2 45.02 72.29 1.00 1.00 1.00
73.00 368.00 Month 3 43 2 34.74 86.80 1.00 1.00 1.00 1.00 461.00
gEVAR/E Day 0 43 2 15.58 39.75 1.00 1.00 1.00 1.00 220.00 Month 1
44 1 40.25 63.63 1.00 1.00 1.00 70.50 296.00 Month 2 45 0 30.38
55.59 1.00 1.00 1.00 68.00 267.00 Month 3 45 0 77.04 205.13 1.00
1.00 1.00 71.00 1135.00 Varilrix gE/Y Day 0 9 1 506.11 1386.23 1.00
1.00 2.00 69.00 4198.00 Month 1 9 1 594.89 1659.25 1.00 1.00 1.00
120.00 5015.00 Month 2 9 1 990.22 2761.19 1.00 1.00 68.00 136.00
8351.00 Month 3 9 1 419.33 1152.89 1.00 1.00 1.00 71.00 3491.00
gEVAR/Y Day 0 10 0 42.30 68.03 1.00 1.00 1.00 67.00 214.00 Month 1
10 0 21.50 45.55 1.00 1.00 1.00 1.00 134.00 Month 2 9 1 77.33
105.58 1.00 1.00 1.00 142.00 274.00 Month 3 10 0 98.40 149.12 1.00
1.00 37.00 141.00 481.00 VAR/E Day 0 44 1 228.18 753.26 1.00 1.00
1.00 138.00 4822.00 Month 1 43 2 205.77 502.36 1.00 1.00 70.00
149.00 3021.00 Month 2 44 1 191.41 509.66 1.00 1.00 34.00 136.00
3158.00 Month 3 45 0 356.69 1417.74 1.00 1.00 70.00 170.00 9496.00
gE/E Day 0 44 1 244.86 491.44 1.00 1.00 71.50 224.50 2300.00 Month
1 42 3 279.14 611.61 1.00 1.00 68.00 225.00 2909.00 Month 2 43 2
236.79 551.54 1.00 1.00 66.00 225.00 2663.00 Month 3 43 2 245.67
489.68 1.00 1.00 71.00 201.00 2491.00 gEVAR/E Day 0 43 2 159.93
381.14 1.00 1.00 1.00 130.00 2072.00 Month 1 44 1 188.82 311.58
1.00 1.00 69.50 217.00 1398.00 Month 2 45 0 223.47 517.02 1.00 1.00
1.00 212.00 2491.00 Month 3 45 0 304.16 520.03 1.00 1.00 143.00
290.00 2487.00 CD8- Pool gE gE/Y Day 0 9 1 30.11 34.57 1.00 1.00
1.00 68.00 68.00 CD40L Month 1 9 1 54.22 112.89 1.00 1.00 1.00
67.00 345.00 Month 2 9 1 565.11 1593.60 1.00 1.00 1.00 68.00
4811.00 Month 3 9 1 16.22 30.21 1.00 1.00 1.00 1.00 70.00 gEVAR/Y
Day 0 10 0 130.10 235.43 1.00 1.00 1.00 206.00 674.00 Month 1 10 0
34.30 65.70 1.00 1.00 1.00 64.00 205.00 Month 2 9 1 31.33 91.00
1.00 1.00 1.00 1.00 274.00 Month 3 10 0 164.20 315.17 1.00 1.00
2.00 204.00 1013.00 VAR/E Day 0 44 1 13.66 31.15 1.00 1.00 1.00
1.00 142.00 Month 1 43 2 9.65 29.29 1.00 1.00 1.00 1.00 153.00
Month 2 43 2 5.19 19.42 1.00 1.00 1.00 1.00 105.00 Month 3 45 0
12.62 31.58 1.00 1.00 1.00 1.00 142.00 gE/E Day 0 44 1 12.32 26.28
1.00 1.00 1.00 1.00 76.00 Month 1 42 3 14.31 27.82 1.00 1.00 1.00
1.00 78.00 Month 2 43 2 17.26 33.86 1.00 1.00 1.00 1.00 146.00
Month 3 43 2 14.16 32.29 1.00 1.00 1.00 1.00 150.00 gEVAR/E Day 0
43 2 7.16 19.48 1.00 1.00 1.00 1.00 71.00 Month 1 44 1 18.95 46.55
1.00 1.00 1.00 1.00 221.00 Month 2 45 0 22.71 43.70 1.00 1.00 1.00
1.00 200.00 Month 3 45 0 45.80 146.31 1.00 1.00 1.00 1.00 780.00
Varilrix gE/Y Day 0 9 1 260.56 651.12 1.00 1.00 68.00 69.00 1992.00
Month 1 9 1 235.78 609.46 1.00 1.00 1.00 120.00 1854.00 Month 2 9 1
291.22 741.43 1.00 1.00 68.00 71.00 2264.00 Month 3 9 1 240.78
644.21 1.00 1.00 1.00 70.00 1954.00 gEVAR/Y Day 0 10 0 21.00 32.21
1.00 1.00 1.00 67.00 69.00 Month 1 10 0 1.00 0.00 1.00 1.00 1.00
1.00 1.00 Month 2 9 1 24.00 49.34 1.00 1.00 1.00 1.00 142.00 Month
3 10 0 62.60 153.10 1.00 1.00 1.00 1.00 481.00 VAR/E Day 0 44 1
15.89 33.39 1.00 1.00 1.00 1.00 152.00 Month 1 43 2 31.95 58.90
1.00 1.00 1.00 68.00 261.00 Month 2 44 1 17.93 45.21 1.00 1.00 1.00
1.00 156.00 Month 3 45 0 17.84 39.12 1.00 1.00 1.00 1.00 152.00
gE/E Day 0 44 1 23.45 53.70 1.00 1.00 1.00 1.50 227.00 Month 1 42 3
25.02 65.40 1.00 1.00 1.00 1.00 363.00 Month 2 43 2 11.16 30.54
1.00 1.00 1.00 1.00 154.00 Month 3 43 2 20.49 44.84 1.00 1.00 1.00
1.00 218.00 gEVAR/E Day 0 43 2 25.58 48.47 1.00 1.00 1.00 1.00
147.00 Month 1 44 1 27.93 55.29 1.00 1.00 1.00 68.00 304.00 Month 2
45 0 12.18 38.15 1.00 1.00 1.00 1.00 225.00 Month 3 45 0 19.80
43.49 1.00 1.00 1.00 1.00 209.00 CD8-IFN.gamma. Pool gE gE/Y Day 0
9 1 15.22 28.26 1.00 1.00 1.00 1.00 68.00 Month 1 9 1 38.44 35.55
1.00 1.00 67.00 68.00 72.00 Month 2 9 1 215.78 472.21 1.00 1.00
68.00 135.00 1464.00 Month 3 9 1 31.11 50.03 1.00 1.00 1.00 66.00
141.00 gEVAR/Y Day 0 10 0 89.40 110.49 1.00 1.00 68.00 143.00
336.00 Month 1 10 0 13.90 27.20 1.00 1.00 1.00 1.00 67.00 Month 2 9
1 31.11 68.87 1.00 1.00 1.00 1.00 205.00 Month 3 10 0 62.40 66.91
1.00 1.00 68.00 76.00 202.00 VAR/E Day 0 44 1 30.77 74.58 1.00 1.00
1.00 1.00 348.00 Month 1 43 2 21.30 50.45 1.00 1.00 1.00 1.00
216.00 Month 2 43 2 38.81 69.55 1.00 1.00 1.00 70.00 284.00 Month 3
45 0 15.13 38.77 1.00 1.00 1.00 1.00 146.00 gE/E Day 0 44 1 12.91
28.23 1.00 1.00 1.00 1.00 110.00 Month 1 42 3 30.40 71.67 1.00 1.00
1.00 1.00 351.00 Month 2 43 2 36.77 73.76 1.00 1.00 1.00 66.00
368.00 Month 3 43 2 31.33 85.07 1.00 1.00 1.00 1.00 461.00 gEVAR/E
Day 0 43 2 12.63 38.39 1.00 1.00 1.00 1.00 220.00 Month 1 44 1
25.36 49.38 1.00 1.00 1.00 35.00 230.00 Month 2 45 0 11.67 25.17
1.00 1.00 1.00 1.00 75.00 Month 3 45 0 58.51 147.37 1.00 1.00 1.00
70.00 851.00 Varilrix gE/Y Day 0 9 1 475.11 1319.18 1.00 1.00 1.00
1.00 3984.00 Month 1 9 1 561.78 1644.43 1.00 1.00 1.00 1.00 4946.00
Month 2 9 1 935.22 2701.84 1.00 1.00 1.00 70.00 8139.00 Month 3 9 1
403.89 1183.38 1.00 1.00 1.00 1.00 3559.00 gEVAR/Y Day 0 10 0 49.30
87.98 1.00 1.00 1.00 68.00 283.00 Month 1 10 0 28.60 58.23 1.00
1.00 1.00 1.00 144.00 Month 2 9 1 61.44 86.91 1.00 1.00 1.00 68.00
208.00 Month 3 10 0 78.20 100.95 1.00 1.00 34.50 147.00 274.00
VAR/E Day 0 44 1 223.32 719.34 1.00 1.00 1.00 134.00 4534.00 Month
1 43 2 208.47 507.58 1.00 1.00 70.00 213.00 3021.00 Month 2 44 1
185.43 499.14 1.00 1.00 1.50 145.50 3085.00 Month 3 45 0 342.33
1408.21 1.00 1.00 31.00 147.00 9423.00 gE/E Day 0 44 1 227.52
482.15 1.00 1.00 1.00 204.50 2216.00 Month 1 42 3 273.88 613.11
1.00 1.00 66.00 225.00 2909.00 Month 2 43 2 228.79 542.44 1.00 1.00
70.00 206.00 2591.00 Month 3 43 2 235.33 490.73 1.00 1.00 69.00
189.00 2491.00 gEVAR/E Day 0 43 2 156.53 390.28 1.00 1.00 1.00
76.00 2000.00 Month 1 44 1 177.48 309.13 1.00 1.00 67.00 217.00
1398.00 Month 2 45 0 220.84 518.38 1.00 1.00 1.00 147.00 2491.00
Month 3 45 0 291.38 515.67 1.00 1.00 92.00 290.00 2418.00 CD8-IL2
Pool gE gE/Y Day 0 9 1 22.67 32.55 1.00 1.00 1.00 62.00 68.00 Month
1 9 1 61.78 111.15 1.00 1.00 1.00 68.00 345.00 Month 2 9 1 557.67
1595.67 1.00 1.00 1.00 68.00 4811.00 Month 3 9 1 31.33 50.21 1.00
1.00 1.00 66.00 141.00 gEVAR/Y Day 0 10 0 136.90 232.25 1.00 1.00
1.00 206.00 674.00 Month 1 10 0 34.30 65.70 1.00 1.00 1.00 64.00
205.00 Month 2 9 1 39.00 114.00 1.00 1.00 1.00 1.00 343.00 Month 3
10 0 170.70 315.26 1.00 1.00 35.50 272.00 1013.00 VAR/E Day 0 44 1
20.30 46.60 1.00 1.00 1.00 1.00 223.00 Month 1 43 2 13.14 36.17
1.00 1.00 1.00 1.00 153.00 Month 2 43 2 16.26 43.32 1.00 1.00 1.00
1.00 212.00 Month 3 45 0 17.49 38.46 1.00 1.00 1.00 1.00 152.00
gE/E Day 0 44 1 16.55 43.92 1.00 1.00 1.00 1.00 212.00 Month 1 42 3
19.29 36.16 1.00 1.00 1.00 1.00 140.00 Month 2 43 2 28.53 54.26
1.00 1.00 1.00 66.00 221.00 Month 3 43 2 16.28 44.66 1.00 1.00 1.00
1.00 229.00 gEVAR/E Day 0 43 2 7.21 19.47 1.00 1.00 1.00 1.00 71.00
Month 1 44 1 25.84 51.64 1.00 1.00 1.00 6.50 221.00 Month 2 45 0
24.18 44.41 1.00 1.00 1.00 3.00 149.00 Month 3 45 0 45.76 148.50
1.00 1.00 1.00 1.00 851.00 Varilrix gE/Y Day 0 9 1 102.78 207.48
1.00 1.00 1.00 69.00 640.00 Month 1 9 1 151.78 359.31 1.00 1.00
1.00 120.00 1098.00 Month 2 9 1 227.78 503.56 1.00 1.00 68.00
136.00 1556.00 Month 3 9 1 93.56 181.12 1.00 1.00 1.00 71.00 559.00
gEVAR/Y Day 0 10 0 14.40 28.25 1.00 1.00 1.00 1.00 69.00 Month 1 10
0 7.60 20.87 1.00 1.00 1.00 1.00 67.00 Month 2 9 1 39.11 60.89 1.00
1.00 1.00 67.00 142.00 Month 3 10 0 62.60 153.10 1.00 1.00 1.00
1.00 481.00 VAR/E Day 0 44 1 99.27 241.07 1.00 1.00 1.00 73.00
1223.00 Month 1 43 2 78.49 122.70 1.00 1.00 66.00 109.00 575.00
Month 2 44 1 97.00 226.12 1.00 1.00 1.00 75.50 1028.00 Month 3 45 0
144.38 452.96 1.00 1.00 1.00 137.00 2994.00 gE/E Day 0 44 1 140.34
240.41 1.00 1.00 33.50 153.00 1271.00 Month 1 42 3 124.21 246.93
1.00 1.00 1.00 145.00 1017.00 Month 2 43 2 99.79 166.75 1.00 1.00
1.00 144.00 863.00 Month 3 43 2 148.19 264.30 1.00 1.00 14.00
145.00 1173.00 gEVAR/E Day 0 43 2 68.81 116.27 1.00 1.00 1.00 77.00
573.00 Month 1 44 1 87.89 122.66 1.00 1.00 68.00 141.00 521.00
Month 2 45 0 113.91 231.46 1.00 1.00 1.00 138.00 1127.00 Month 3 45
0 163.44 232.53 1.00 1.00 71.00 215.00 967.00 CD8-TNF.alpha. Pool
gE gE/Y Day 0 9 1 23.33 33.50 1.00 1.00 1.00 68.00 68.00 Month 1 9
1 1.00 0.00 1.00 1.00 1.00 1.00 1.00 Month 2 9 1 15.89 44.67 1.00
1.00 1.00 1.00 135.00 Month 3 9 1 23.89 49.04 1.00 1.00 1.00 1.00
141.00 gEVAR/Y Day 0 10 0 35.00 65.20 1.00 1.00 1.00 68.00 201.00
Month 1 10 0 1.00 0.00 1.00 1.00 1.00 1.00 1.00 Month 2 9 1 8.56
22.67 1.00 1.00 1.00 1.00 69.00 Month 3 10 0 8.00 21.10 1.00 1.00
1.00 1.00 68.00 VAR/E Day 0 44 1 32.20 75.14 1.00 1.00 1.00 1.50
348.00 Month 1 43 2 33.05 63.48 1.00 1.00 1.00 72.00 288.00 Month 2
43 2 31.40 67.10 1.00 1.00 1.00 12.00 284.00 Month 3 45 0 19.80
48.19 1.00 1.00 1.00 1.00 207.00 gE/E Day 0 44 1 18.80 44.11 1.00
1.00 1.00 1.00 221.00 Month 1 42 3 32.60 77.37 1.00 1.00 1.00 37.00
422.00 Month 2 43 2 30.02 47.35 1.00 1.00 1.00 66.00 221.00 Month 3
43 2 24.53 67.55 1.00 1.00 1.00 1.00 306.00 gEVAR/E Day 0 43 2
15.79 40.14 1.00 1.00 1.00 1.00 220.00 Month 1 44 1 28.91 57.44
1.00 1.00 1.00 33.50 230.00 Month 2 45 0 21.13 41.06 1.00 1.00 1.00
1.00 149.00 Month 3 45 0 52.49 124.92 1.00 1.00 1.00 69.00 709.00
Varilrix gE/Y Day 0 9 1 459.33 1297.02 1.00 1.00 1.00 2.00 3913.00
Month 1 9 1 535.67 1526.03 1.00 1.00 1.00 70.00 4603.00 Month 2 9 1
928.11 2730.86 1.00 1.00 1.00 67.00 8210.00 Month 3 9 1 373.22
1090.66 1.00 1.00 1.00 1.00 3281.00 gEVAR/Y Day 0 10 0 35.60 68.51
1.00 1.00 1.00 67.00 214.00 Month 1 10 0 21.50 45.55 1.00 1.00 1.00
1.00 134.00 Month 2 9 1 69.33 102.76 1.00 1.00 1.00 70.00 274.00
Month 3 10 0 44.20 77.20 1.00 1.00 1.00 73.00 215.00 VAR/E Day 0 44
1 200.27 729.98 1.00 1.00 1.00 75.00 4678.00 Month 1 43 2 168.60
469.56 1.00 1.00 34.00 141.00 2805.00 Month 2 44 1 159.45 432.79
1.00 1.00 1.00 129.50 2717.00 Month 3 45 0 309.73 1310.43 1.00 1.00
1.00 133.00 8765.00 gE/E Day 0 44 1 201.23 441.88 1.00 1.00 1.00
150.50 2057.00 Month 1 42 3 230.05 531.94 1.00 1.00 47.50 225.00
2545.00 Month 2 43 2 221.44 535.05 1.00 1.00 66.00 225.00 2591.00
Month 3 43 2 189.44 402.52 1.00 1.00 61.00 189.00 2125.00 gEVAR/E
Day 0 43 2 133.88 332.42 1.00 1.00 1.00 72.00 1712.00 Month 1 44 1
172.50 306.32 1.00 1.00 67.00 212.50 1398.00 Month 2 45 0 190.29
470.52 1.00 1.00 1.00 147.00 2340.00 Month 3 45 0 255.16 490.59
1.00 1.00 71.00 215.00 2279.00 gE/Y = gE-AS1/18-30 years gEVAR/Y =
gE-AS1 + Varilrix/18-30 years VAR/E = Varilrix/50-70 years gE/E =
gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70 years N =
number of subjects with available results N miss. = number of
subjects with missing results SD = Standard Deviation Min, Max =
Minimum, Maximum Q1, Q3 = First, Third quartile
TABLE-US-00016 TABLE C.2 Intracellular Cytokine Staining (ICS):
Inferential statistics: P- values from Kruskal-Wallis Tests for CD4
T cells at each time point (Total Vaccinated Cohort) Groups P_value
at P_value at P_value at P_value at T cells compared Antigen Test
day 0 Month 1 Month 2 Month 3 CD4 VAR\E and Pool gE ALL DOUBLES
0.5025 0.0000 0.0015 0.0000 gEVAR\E CD40L 0.4448 0.0000 0.0004
0.0000 IFN.gamma. 0.5900 0.0001 0.0956 0.0000 IL2 0.6415 0.0000
0.0001 0.0000 TNF.alpha. 0.2634 0.0000 0.0019 0.0000 Varilrix ALL
DOUBLES 0.7118 0.1489 0.3148 0.0000 CD40L 0.6488 0.1664 0.2609
0.0000 IFN.gamma. 0.3602 0.2905 0.2277 0.0000 IL2 0.4880 0.1442
0.2406 0.0000 TNF.alpha. 0.8631 0.2624 0.2455 0.0000 VAR\E and Pool
gE ALL DOUBLES 0.9764 0.0004 0.0100 0.0000 gE\E CD40L 0.9765 0.0003
0.0026 0.0000 IFN.gamma. 0.9665 0.0228 0.2961 0.0000 IL2 0.7183
0.0001 0.0035 0.0000 TNF.alpha. 0.9026 0.0069 0.0053 0.0000
Varilrix ALL DOUBLES 0.9069 0.9965 0.8552 0.0002 CD40L 0.8904
0.9790 0.9155 0.0002 IFN.gamma. 0.8806 0.5797 0.6868 0.0010 IL2
0.9601 0.9860 0.9054 0.0003 TNF.alpha. 0.6073 0.9719 0.8154 0.0016
gE\E and Pool gE ALL DOUBLES 0.4951 0.1777 0.5702 0.4832 gEVAR\E
CD40L 0.3731 0.2215 0.5312 0.5368 IFN.gamma. 0.7732 0.2331 0.5958
0.8576 IL2 0.9406 0.3059 0.4181 0.5069 TNF.alpha. 0.3949 0.2039
0.5613 0.3287 Varilrix ALL DOUBLES 0.8469 0.1876 0.2409 0.2687
CD40L 0.9803 0.1980 0.2060 0.2277 IFN.gamma. 0.7520 0.2103 0.1205
0.2182 IL2 0.7211 0.2135 0.2375 0.3045 TNF.alpha. 0.8118 0.2134
0.1817 0.2778 VAR/E = Varilrix/50-70 years gE/E = gE-AS1/50-70
years gEVAR/E = gE-AS1 + Varilrix/50-70 years
TABLE-US-00017 SUPPLEMENTARY TABLE C.2 Intracellular Cytokine
Staining (ICS): Inferential statistics: P-values from
Kruskal-Wallis Tests for CD8 T cells at each time point (Total
Vaccinated Cohort) Groups P_value at P_value at P_value at P_value
at T cells compared Antigen Test day 0 Month 1 Month 2 Month 3 CD8
VAR\E and Pool gE ALL DOUBLES 0.1477 0.4418 0.8141 0.2762 gEVAR\E
CD40L 0.2897 0.2513 0.0126 0.3511 IFN.gamma. 0.1695 0.4069 0.0478
0.0478 IL2 0.2705 0.1316 0.2008 0.5872 TNF.alpha. 0.2968 0.7017
0.6470 0.0947 Varilrix ALL DOUBLES 0.9267 0.7605 0.9651 0.1197
CD40L 0.6260 0.9111 0.6512 0.8826 IFN.gamma. 0.9846 0.9611 0.9225
0.1009 IL2 0.7027 0.6963 0.4626 0.1181 TNF.alpha. 0.9047 0.4655
0.9929 0.1639 VAR\E and Pool gE ALL DOUBLES 0.4117 0.9608 0.4570
0.9320 gE\E CD40L 0.7891 0.2636 0.0315 0.7302 IFN.gamma. 0.4922
0.5672 0.7960 0.3690 IL2 0.6092 0.2137 0.1416 0.6416 TNF.alpha.
0.5891 0.8828 0.4633 0.9530 Varilrix ALL DOUBLES 0.2336 0.9168
0.4792 0.6436 CD40L 0.5969 0.3443 0.6968 0.8133 IFN.gamma. 0.3606
0.9342 0.3019 0.5406 IL2 0.1743 0.6509 0.2577 0.4652 TNF.alpha.
0.3405 0.7627 0.3869 0.5577 gE\E and Pool gE ALL DOUBLES 0.4942
0.3322 0.2975 0.3120 gEVAR\E CD40L 0.1831 0.9898 0.6047 0.5555
IFN.gamma. 0.4515 0.8129 0.0948 0.2325 IL2 0.6171 0.7224 0.8439
0.3147 TNF.alpha. 0.6064 0.7472 0.2571 0.1078 Varilrix ALL DOUBLES
0.2524 0.8479 0.4410 0.2783 CD40L 0.9594 0.3385 0.9095 0.9433
IFN.gamma. 0.3465 0.9277 0.3691 0.2849 IL2 0.2333 0.5263 0.7101
0.4173 TNF.alpha. 0.4678 0.7167 0.3198 0.4684 VAR/E =
Varilrix/50-70 years gE/E = gE-AS1/50-70 years gEVAR/E = gE-AS1 +
Varilrix/50-70 years
TABLE-US-00018 TABLE C.3 Intracellular Cytokine Staining (ICS):
Descriptive Statistics on CD4 T cells at POST-PRE (Total vaccinated
Cohort) Test Antigen Group Timing N N miss. Mean SD Min Q1 Median
Q3 Max CD4-ALL Pool gE gE/Y Month 1 9 1 1170.33 1640.79 -80.00
230.00 434.00 1332.00 5096.00 DOUBLES Month 2 9 1 1574.11 1884.90
256.00 496.00 723.00 1285.00 5428.00 Month 3 9 1 2526.44 1927.96
442.00 1329.00 2233.00 2647.00 6852.00 gEVAR/Y Month 1 10 0 926.50
846.70 -348.00 393.00 865.50 1363.00 2752.00 Month 2 9 1 1308.56
846.35 -233.00 845.00 1681.00 1995.00 2178.00 Month 3 10 0 3045.30
1361.87 1288.00 1969.00 2830.50 4533.00 5057.00 VAR/E Month 1 42 3
147.90 460.77 -436.00 -49.00 25.50 155.00 1898.00 Month 2 43 2
-84.23 947.38 -5979.00 -60.00 35.00 179.00 658.00 Month 3 44 1
173.16 1441.28 -5191.00 -128.00 0.00 146.00 6263.00 gE/E Month 1 42
3 691.74 1008.97 -163.00 135.00 310.00 826.00 4210.00 Month 2 43 2
352.00 515.95 -337.00 39.00 206.00 485.00 1974.00 Month 3 42 3
3118.48 2537.47 -112.00 1324.00 2344.50 4620.00 11479.00 gEVAR/E
Month 1 44 1 616.02 505.69 -184.00 165.00 597.50 902.00 2377.00
Month 2 45 0 303.13 389.20 -549.00 52.00 266.00 515.00 1234.00
Month 3 45 0 2712.02 2508.85 -542.00 925.00 1601.00 4223.00
10467.00 Varilrix gE/Y Month 1 9 1 256.89 842.23 -811.00 -214.00
-87.00 712.00 1612.00 Month 2 9 1 610.22 651.41 -72.00 129.00
361.00 831.00 1904.00 Month 3 9 1 770.56 891.01 -787.00 548.00
718.00 1165.00 2343.00 gEVAR/Y Month 1 10 0 -98.40 1180.58 -1610.00
-647.00 -31.50 75.00 2732.00 Month 2 9 1 1547.67 993.79 225.00
797.00 1286.00 2179.00 3188.00 Month 3 10 0 2181.30 1781.51 151.00
698.00 1867.00 2861.00 6210.00 VAR/E Month 1 42 3 366.95 798.30
-663.00 0.00 173.00 536.00 4145.00 Month 2 43 2 169.07 569.41
-2100.00 -39.00 197.00 325.00 1608.00 Month 3 44 1 362.93 662.91
-2100.00 54.00 267.50 633.50 1906.00 gE/E Month 1 42 3 524.74
946.86 -660.00 0.00 229.00 734.00 4178.00 Month 2 43 2 95.30 633.65
-1405.00 -205.00 18.00 254.00 1961.00 Month 3 42 3 1533.69 1557.01
-600.00 528.00 1090.00 2181.00 7044.00 gEVAR/E Month 1 44 1 664.14
765.08 -555.00 184.50 575.00 1109.00 3364.00 Month 2 45 0 451.04
652.53 -397.00 34.00 244.00 639.00 2601.00 Month 3 45 0 1973.73
1577.30 70.00 918.00 1480.00 2659.00 6575.00 CD4-CD40L Pool gE gE/Y
Month 1 9 1 1142.67 1567.96 -77.00 290.00 434.00 1272.00 4889.00
Month 2 9 1 1519.78 1786.74 336.00 465.00 680.00 1369.00 5120.00
Month 3 9 1 2363.22 1772.92 375.00 1303.00 1870.00 2323.00 6282.00
gEVAR/Y Month 1 10 0 868.50 852.69 -348.00 381.00 813.00 1265.00
2782.00 Month 2 9 1 1278.44 841.17 -233.00 691.00 1620.00 2027.00
2048.00 Month 3 10 0 2915.50 1350.46 1150.00 1816.00 2785.50
4273.00 5088.00 VAR/E Month 1 42 3 141.24 434.95 -487.00 -22.00
44.00 161.00 1837.00 Month 2 43 2 -80.09 893.99 -5627.00 -74.00
26.00 195.00 601.00 Month 3 44 1 169.66 1401.60 -4969.00 -100.00
6.50 136.00 6071.00 gE/E Month 1 42 3 691.05 975.64 -183.00 154.00
323.00 814.00 4153.00 Month 2 43 2 367.37 493.31 -306.00 64.00
222.00 546.00 1976.00 Month 3 42 3 3036.88 2447.00 -189.00 1342.00
2320.00 4568.00 10509.00 gEVAR/E Month 1 44 1 603.75 496.38 -112.00
183.00 583.50 878.50 2268.00 Month 2 45 0 308.96 364.06 -493.00
115.00 266.00 515.00 1185.00 Month 3 45 0 2692.24 2497.48 -397.00
918.00 1652.00 4189.00 10417.00 Varilrix gE/Y Month 1 9 1 236.33
828.70 -790.00 -212.00 -62.00 634.00 1594.00 Month 2 9 1 581.89
584.04 -74.00 217.00 361.00 781.00 1587.00 Month 3 9 1 722.89
892.40 -737.00 210.00 747.00 1091.00 2284.00 gEVAR/Y Month 1 10 0
-122.90 1156.16 -1604.00 -613.00 -114.50 75.00 2676.00 Month 2 9 1
1519.67 1018.54 259.00 684.00 1228.00 2137.00 3210.00 Month 3 10 0
2054.60 1759.67 36.00 554.00 1840.50 2421.00 6057.00 VAR/E Month 1
42 3 355.62 780.99 -623.00 25.00 150.50 529.00 4128.00 Month 2 43 2
160.72 567.36 -2024.00 -32.00 158.00 349.00 1631.00 Month 3 44 1
351.45 663.33 -2024.00 4.00 199.00 616.50 1972.00 gE/E Month 1 42 3
509.52 852.27 -679.00 -5.00 213.50 747.00 3242.00 Month 2 43 2
109.05 630.36 -1378.00 -181.00 27.00 257.00 2004.00 Month 3 42 3
1501.02 1546.41 -881.00 562.00 1036.50 2095.00 7128.00 gEVAR/E
Month 1 44 1 661.55 758.32 -487.00 155.50 593.50 1066.50 3519.00
Month 2 45 0 457.07 652.72 -411.00 33.00 290.00 580.00 2646.00
Month 3 45 0 1931.53 1542.71 71.00 916.00 1427.00 2686.00 6529.00
CD4-IFN.gamma. Pool gE gE/Y Month 1 9 1 855.89 1270.64 -83.00 56.00
498.00 823.00 3836.00 Month 2 9 1 1152.67 1414.76 192.00 402.00
511.00 1087.00 4569.00 Month 3 9 1 1841.44 1805.38 254.00 894.00
1227.00 1892.00 6156.00 gEVAR/Y Month 1 10 0 646.20 866.49 -241.00
194.00 402.00 739.00 2897.00 Month 2 9 1 898.89 743.23 -203.00
367.00 1064.00 1602.00 1789.00 Month 3 10 0 2108.30 1216.76 920.00
1089.00 1854.50 2667.00 5000.00 VAR/E Month 1 42 3 104.52 323.14
-231.00 -17.00 29.50 84.00 1809.00 Month 2 43 2 -25.56 505.62
-3122.00 -24.00 33.00 117.00 561.00 Month 3 44 1 99.75 731.24
-2787.00 -32.00 11.00 88.50 3037.00 gE/E Month 1 42 3 395.14 676.51
-72.00 45.00 174.50 333.00 3116.00 Month 2 43 2 161.98 330.61
-305.00 5.00 54.00 219.00 1539.00 Month 3 42 3 1583.48 1568.34
-47.00 491.00 1224.00 2077.00 6308.00 gEVAR/E Month 1 44 1 350.95
333.21 -34.00 91.00 270.50 554.00 1349.00 Month 2 45 0 172.09
273.80 -186.00 8.00 82.00 274.00 1062.00 Month 3 45 0 1392.40
1301.10 -29.00 484.00 875.00 1961.00 5786.00 Varilrix gE/Y Month 1
9 1 196.44 699.90 -663.00 -156.00 -144.00 431.00 1472.00 Month 2 9
1 428.22 503.10 -132.00 84.00 332.00 743.00 1342.00 Month 3 9 1
507.00 696.10 -781.00 196.00 471.00 831.00 1563.00 gEVAR/Y Month 1
10 0 -25.40 1073.83 -1588.00 -456.00 -20.00 106.00 2599.00 Month 2
9 1 1246.56 985.49 88.00 467.00 945.00 1661.00 3056.00 Month 3 10 0
1768.90 1593.80 168.00 634.00 1485.50 2136.00 5549.00 VAR/E Month 1
42 3 309.07 689.54 -701.00 16.00 187.50 396.00 3829.00 Month 2 43 2
116.07 496.80 -1934.00 -74.00 134.00 283.00 1515.00 Month 3 44 1
254.25 539.74 -1750.00 23.00 161.50 445.00 1861.00 gE/E Month 1 42
3 405.48 723.30 -631.00 -25.00 204.00 515.00 2775.00 Month 2 43 2
33.23 464.33 -1258.00 -102.00 42.00 204.00 1236.00 Month 3 42 3
1042.86 1155.33 -529.00 402.00 805.00 1590.00 6357.00 gEVAR/E Month
1 44 1 548.59 714.90 -621.00 111.00 374.50 757.50 3276.00 Month 2
45 0 399.91 618.77 -454.00 47.00 231.00 474.00 2730.00 Month 3 45 0
1382.96 1099.42 -1.00 652.00 1088.00 1679.00 4831.00 CD4-IL2 Pool
gE gE/Y Month 1 9 1 1093.00 1486.37 -40.00 228.00 465.00 1118.00
4669.00 Month 2 9 1 1495.67 1762.08 316.00 524.00 623.00 1301.00
5044.00 Month 3 9 1 2151.22 1569.05 344.00 1086.00 2010.00 2510.00
5502.00 gEVAR/Y Month 1 10 0 750.70 647.88 -348.00 466.00 613.00
1063.00 2048.00 Month 2 9 1 1167.22 708.40 -79.00 801.00 1192.00
1802.00 1974.00 Month 3 10 0 2510.60 1142.73 1026.00 1562.00
2307.50 3545.00 4153.00 VAR/E Month 1 42 3 106.40 368.83 -361.00
-83.00 54.00 126.00 1897.00 Month 2 43 2 -85.70 860.78 -5429.00
-81.00 0.00 159.00 634.00 Month 3 44 1 125.25 1203.10 -4856.00
-80.50 0.00 137.00 4576.00 gE/E Month 1 42 3 617.10 907.63 -49.00
128.00 312.50 692.00 3736.00 Month 2 43 2 314.26 460.49 -271.00
25.00 189.00 389.00 1723.00 Month 3 42 3 2711.76 2304.96 0.00
1008.00 2082.00 4222.00 10315.00 gEVAR/E Month 1 44 1 543.70 465.89
-111.00 104.00 508.50 826.00 1920.00 Month 2 45 0 300.42 336.69
-460.00 103.00 300.00 429.00 1088.00 Month 3 45 0 2385.11 2307.17
-460.00 819.00 1478.00 3888.00 9485.00 Varilrix gE/Y Month 1 9 1
150.78 735.34 -812.00 -271.00 -77.00 664.00 1263.00 Month 2 9 1
540.56 597.57 -125.00 211.00 316.00 712.00 1721.00 Month 3 9 1
609.78 735.25 -565.00 377.00 576.00 895.00 1986.00 gEVAR/Y Month 1
10 0 -171.70 949.81 -1498.00 -545.00 -111.00 135.00 1991.00 Month 2
9 1 1396.11 925.12 443.00 678.00 1111.00 1980.00 3192.00 Month 3 10
0 1717.20 1413.72 133.00 532.00 1356.50 2606.00 4739.00 VAR/E Month
1 42 3 290.62 714.95 -689.00 5.00 161.00 401.00 3784.00 Month 2 43
2 121.05 534.68 -2122.00 -83.00 134.00 369.00 1492.00 Month 3 44 1
257.48 599.43 -1964.00 -17.00 196.00 504.50 1857.00 gE/E Month 1 42
3 438.14 817.33 -695.00 -21.00 173.00 644.00 3233.00 Month 2 43 2
71.42 575.91 -1320.00 -274.00 69.00 255.00 1678.00 Month 3 42 3
1254.76 1389.73 -548.00 382.00 827.50 1798.00 6315.00 gEVAR/E Month
1 44 1 586.45 695.20 -363.00 169.50 437.50 858.00 3292.00 Month 2
45 0 420.22 582.63 -416.00 78.00 185.00 545.00 2287.00 Month 3 45 0
1648.53 1355.42 71.00 742.00 1148.00 2418.00 5462.00 CD4-TNF.alpha.
Pool gE gE/Y Month 1 9 1 560.22 934.14 12.00 65.00 123.00 756.00
2903.00 Month 2 9 1 791.78 1375.33 3.00 149.00 315.00 623.00
4378.00 Month 3 9 1 1324.00 1590.20 206.00 415.00 831.00 1105.00
5212.00 gEVAR/Y Month 1 10 0 532.30 452.83 -11.00 253.00 503.50
651.00 1572.00 Month 2 9 1 715.00 462.65 38.00 503.00 712.00
1039.00 1314.00 Month 3 10 0 1653.00 811.66 465.00 1195.00 1850.50
1931.00 2902.00 VAR/E Month 1 42 3 51.69 229.81 -470.00 -57.00
41.00 115.00 1215.00 Month 2 43 2 -86.14 688.49 -4412.00 -46.00
2.00 126.00 254.00 Month 3 44 1 77.75 960.07 -3944.00 -80.00 1.00
94.00 4212.00 gE/E Month 1 42 3 325.40 519.68 -165.00 0.00 161.50
419.00 2302.00 Month 2 43 2 191.19 319.19 -246.00 0.00 119.00
225.00 1367.00 Month 3 42 3 1815.83 1594.15 -152.00 690.00 1389.50
2762.00 7491.00 gEVAR/E Month 1 44 1 275.16 261.28 -156.00 57.50
233.50 480.00 1045.00 Month 2 45 0 164.93 267.03 -494.00 26.00
126.00 316.00 675.00 Month 3 45 0 1508.64 1548.74 -433.00 508.00
875.00 2193.00 6403.00 Varilrix gE/Y Month 1 9 1 57.67 443.72
-683.00 -219.00 47.00 528.00 634.00 Month 2 9 1 450.11 411.03
-101.00 184.00 411.00 513.00 1364.00 Month 3 9 1 374.89 664.84
-945.00 113.00 404.00 637.00 1536.00 gEVAR/Y Month 1 10 0 -156.30
634.57 -1203.00 -365.00 -107.50 217.00 941.00 Month 2 9 1 1047.56
757.92 179.00 535.00 765.00 1585.00 2566.00 Month 3 10 0 1200.00
918.82 -123.00 626.00 1214.50 1693.00 3132.00 VAR/E Month 1 42 3
250.64 695.21 -805.00 -33.00 138.50 281.00 3707.00 Month 2 43 2
110.26 519.34 -1984.00 -121.00 103.00 294.00 1491.00 Month 3 44 1
210.23 548.98 -1984.00 -10.00 167.50 423.50 1806.00 gE/E Month 1 42
3 298.93 637.78 -566.00 -45.00 146.50 405.00 2494.00 Month 2 43 2
29.81 538.43 -1359.00 -171.00 13.00 150.00 1845.00 Month 3 42 3
967.05 1085.98 -609.00 239.00 707.50 1502.00 4441.00 gEVAR/E Month
1 44 1 405.30 604.26 -432.00 90.00 304.50 554.50 2871.00 Month 2 45
0 293.78 495.54 -347.00 -34.00 167.00 324.00 2242.00 Month 3 45 0
1229.38 1061.80 -73.00 492.00 913.00 1565.00 4380.00 gE/Y =
gE-AS1/18-30 years gEVAR/Y = gE-AS1 + Varilrix/18-30 years VAR/E =
Varilrix/50-70 years gE/E = gE-AS1/50-70 years gEVAR/E = gE-AS1 +
Varilrix/50-70 years N = number of subjects with available results
N miss. = number of subjects with missing results SD = Standard
Deviation Min, Max = Minimum, Maximum Q1, Q3 = First, Third
quartile
TABLE-US-00019 SUPPLEMENTARY TABLE C.3 Intracellular Cytokine
Staining (ICS): Descriptive Statistics on CD8 T cells at POST-PRE
(Total vaccinated Cohort) Test Antigen Group POST N N miss. Mean SD
Min Q1 Median Q3 Max CD8-ALL Pool gE gE/Y Month 1 9 1 24.00 136.52
-136.00 -61.00 0.00 67.00 344.00 DOUBLES Month 2 9 1 549.89 1599.52
-67.00 -2.00 0.00 69.00 4810.00 Month 3 9 1 0.89 83.03 -136.00
-61.00 0.00 65.00 140.00 gEVAR/Y Month 1 10 0 -116.70 198.88
-537.00 -139.00 -34.00 0.00 63.00 Month 2 9 1 -128.67 170.57
-413.00 -205.00 -68.00 0.00 0.00 Month 3 10 0 26.80 117.89 -142.00
-68.00 0.00 69.00 271.00 VAR/E Month 1 42 3 -13.38 97.87 -278.00
-65.00 0.00 0.00 210.00 Month 2 42 3 -3.50 76.50 -222.00 -22.00
0.00 0.00 209.00 Month 3 44 1 -10.48 107.47 -347.00 -11.00 0.00
0.00 285.00 gE/E Month 1 42 3 10.45 95.31 -211.00 0.00 0.00 1.00
421.00 Month 2 43 2 22.51 95.28 -220.00 0.00 0.00 70.00 367.00
Month 3 42 3 7.29 88.24 -211.00 0.00 0.00 0.00 460.00 gEVAR/E Month
1 42 3 26.12 69.05 -71.00 0.00 0.00 68.00 295.00 Month 2 43 2 16.16
62.56 -147.00 0.00 0.00 67.00 201.00 Month 3 43 2 65.00 217.39
-219.00 0.00 0.00 70.00 1134.00 Varilrix gE/Y Month 1 9 1 88.78
274.69 -67.00 -1.00 0.00 0.00 817.00 Month 2 9 1 484.11 1376.23
-1.00 0.00 1.00 68.00 4153.00 Month 3 9 1 -86.78 246.23 -707.00
-68.00 0.00 0.00 135.00 gEVAR/Y Month 1 10 0 -20.80 71.94 -141.00
-66.00 0.00 0.00 133.00 Month 2 9 1 30.44 143.10 -213.00 -66.00
0.00 139.00 273.00 Month 3 10 0 56.10 163.61 -213.00 0.00 35.00
140.00 414.00 VAR/E Month 1 42 3 -31.67 340.22 -1801.00 -71.00 0.00
74.00 491.00 Month 2 43 2 -52.51 298.58 -1664.00 -68.00 0.00 68.00
362.00 Month 3 44 1 131.75 725.55 -611.00 -26.00 0.00 104.50
4674.00 gE/E Month 1 42 3 59.62 265.24 -296.00 -72.00 0.00 81.00
1015.00 Month 2 43 2 -10.51 382.79 -1736.00 -42.00 0.00 71.00
1242.00 Month 3 42 3 28.60 190.71 -423.00 -71.00 0.00 72.00 597.00
gEVAR/E Month 1 42 3 5.90 351.50 -1924.00 -1.00 0.00 73.00 694.00
Month 2 43 2 15.98 365.42 -1594.00 -70.00 0.00 143.00 1213.00 Month
3 43 2 129.19 225.91 -149.00 0.00 16.00 265.00 916.00 CD8- Pool gE
gE/Y Month 1 9 1 24.11 127.75 -67.00 -61.00 0.00 0.00 344.00 CD40L
Month 2 9 1 535.00 1604.13 -67.00 0.00 0.00 0.00 4810.00 Month 3 9
1 -13.89 55.72 -67.00 -67.00 0.00 0.00 69.00 gEVAR/Y Month 1 10 0
-95.80 189.16 -469.00 -139.00 0.00 0.00 63.00 Month 2 9 1 -113.11
179.39 -413.00 -205.00 0.00 0.00 0.00 Month 3 10 0 34.10 138.26
-210.00 0.00 0.00 69.00 339.00 VAR/E Month 1 42 3 -4.40 43.17
-141.00 0.00 0.00 0.00 152.00 Month 2 42 3 -7.29 37.64 -141.00 0.00
0.00 0.00 104.00 Month 3 44 1 -0.77 37.03 -73.00 0.00 0.00 0.00
102.00 gE/E Month 1 42 3 1.45 39.39 -75.00 0.00 0.00 0.00 76.00
Month 2 43 2 6.33 45.80 -75.00 0.00 0.00 0.00 145.00 Month 3 42 3
-1.93 36.10 -75.00 0.00 0.00 0.00 72.00 gEVAR/E Month 1 42 3 10.86
48.91 -70.00 0.00 0.00 0.00 220.00 Month 2 43 2 16.56 45.02 -70.00
0.00 0.00 65.00 145.00 Month 3 43 2 40.72 152.63 -70.00 0.00 0.00
0.00 779.00 Varilrix gE/Y Month 1 9 1 -24.78 56.16 -138.00 -67.00
0.00 0.00 51.00 Month 2 9 1 30.67 102.65 -68.00 -1.00 0.00 69.00
272.00 Month 3 9 1 -19.78 46.08 -73.00 -67.00 0.00 0.00 68.00
gEVAR/Y Month 1 10 0 -20.00 32.21 -68.00 -66.00 0.00 0.00 0.00
Month 2 9 1 0.78 68.53 -68.00 -66.00 0.00 0.00 141.00 Month 3 10 0
41.60 134.83 -68.00 0.00 0.00 0.00 414.00 VAR/E Month 1 42 3 16.17
63.26 -72.00 0.00 0.00 0.00 260.00 Month 2 43 2 2.09 48.77 -73.00
0.00 0.00 0.00 153.00 Month 3 44 1 2.34 43.59 -73.00 0.00 0.00 0.00
146.00 gE/E Month 1 42 3 2.14 80.48 -149.00 0.00 0.00 0.00 362.00
Month 2 43 2 -11.23 59.14 -218.00 0.00 0.00 0.00 152.00 Month 3 42
3 -1.93 76.17 -226.00 0.00 0.00 0.00 217.00 gEVAR/E Month 1 42 3
3.05 51.88 -135.00 0.00 0.00 0.00 157.00 Month 2 43 2 -12.88 47.07
-145.00 0.00 0.00 0.00 78.00 Month 3 43 2 -4.91 46.80 -145.00 0.00
0.00 0.00 73.00 CD8-IFN.gamma. Pool gE gE/Y Month 1 9 1 23.22 45.52
-61.00 0.00 4.00 66.00 67.00 Month 2 9 1 200.56 477.92 -61.00 0.00
67.00 133.00 1463.00 Month 3 9 1 15.89 65.31 -67.00 0.00 0.00 65.00
140.00 gEVAR/Y Month 1 10 0 -75.50 120.41 -335.00 -142.00 -34.00
0.00 63.00 Month 2 9 1 -68.11 59.39 -142.00 -131.00 -67.00 0.00
0.00 Month 3 10 0 -27.00 56.93 -134.00 -68.00 0.00 0.00 67.00 VAR/E
Month 1 42 3 -15.52 75.76 -278.00 0.00 0.00 0.00 193.00 Month 2 42
3 2.43 62.84 -139.00 0.00 0.00 0.00 192.00 Month 3 44 1 -15.32
79.50 -347.00 0.00 0.00 0.00 75.00 gE/E Month 1 42 3 16.93 72.52
-109.00 0.00 0.00 0.00 282.00 Month 2 43 2 23.58 84.49 -109.00 0.00
0.00 65.00 367.00 Month 3 42 3 18.57 70.71 -66.00 0.00 0.00 0.00
392.00 gEVAR/E Month 1 42 3 13.55 62.27 -146.00 0.00 0.00 0.00
229.00 Month 2 43 2 -0.47 40.58 -147.00 0.00 0.00 0.00 74.00 Month
3 43 2 48.56 146.44 -144.00 0.00 0.00 68.00 781.00 Varilrix gE/Y
Month 1 9 1 86.67 344.40 -284.00 0.00 0.00 0.00 962.00 Month 2 9 1
460.11 1388.68 -215.00 0.00 0.00 66.00 4155.00 Month 3 9 1 -71.22
165.93 -425.00 0.00 0.00 0.00 68.00 gEVAR/Y Month 1 10 0 -20.70
71.64 -139.00 -67.00 0.00 0.00 133.00 Month 2 9 1 6.78 136.42
-282.00 -1.00 0.00 67.00 204.00 Month 3 10 0 28.90 155.44 -282.00
0.00 0.50 146.00 273.00 VAR/E Month 1 42 3 -23.81 304.92 -1513.00
0.00 0.00 73.00 561.00 Month 2 43 2 -53.65 284.04 -1449.00 0.00
0.00 65.00 289.00 Month 3 44 1 121.93 758.36 -610.00 -0.50 0.00
73.00 4889.00 gE/E Month 1 42 3 74.10 267.93 -296.00 -2.00 0.00
71.00 1088.00 Month 2 43 2 -4.00 372.80 -1652.00 -66.00 0.00 68.00
1170.00 Month 3 42 3 37.74 175.85 -339.00 -3.00 0.00 74.00 670.00
gEVAR/E Month 1 42 3 -0.81 360.67 -1999.00 0.00 0.00 73.00 694.00
Month 2 43 2 16.63 341.39 -1452.00 -66.00 0.00 72.00 1213.00 Month
3 43 2 119.21 197.20 -284.00 0.00 16.00 221.00 659.00 CD8-IL2 Pool
gE gE/Y Month 1 9 1 39.11 127.97 -67.00 -61.00 0.00 67.00 344.00
Month 2 9 1 535.00 1604.48 -67.00 -61.00 0.00 67.00 4810.00 Month 3
9 1 8.67 71.13 -67.00 -61.00 0.00 65.00 140.00 gEVAR/Y Month 1 10 0
-102.60 186.54 -469.00 -139.00 0.00 0.00 63.00 Month 2 9 1 -113.00
162.73 -413.00 -205.00 0.00 0.00 0.00 Month 3 10 0 33.80 125.98
-142.00 0.00 0.00 69.00 339.00 VAR/E Month 1 42 3 -7.79 57.52
-148.00 0.00 0.00 0.00 152.00 Month 2 42 3 -2.90 68.64 -222.00 0.00
0.00 0.00 211.00 Month 3 44 1 -4.02 64.96 -222.00 0.00 0.00 0.00
151.00 gE/E Month 1 42 3 2.00 52.68 -211.00 0.00 0.00 0.00 139.00
Month 2 43 2 13.28 64.17 -211.00 0.00 0.00 65.00 218.00 Month 3 42
3 -4.19 59.14 -211.00 0.00 0.00 0.00 228.00 gEVAR/E Month 1 42 3
19.67 59.09 -70.00 0.00 0.00 11.00 220.00 Month 2 43 2 18.05 48.05
-70.00 0.00 0.00 65.00 148.00 Month 3 43 2 40.63 153.41 -66.00 0.00
0.00 0.00 850.00 Varilrix gE/Y Month 1 9 1 49.00 156.22 -67.00 0.00
0.00 0.00 458.00 Month 2 9 1 125.00 298.57 -1.00 0.00 0.00 70.00
916.00 Month 3 9 1 -9.22 73.85 -81.00 -68.00 0.00 0.00 136.00
gEVAR/Y Month 1 10 0 -6.80 37.82 -68.00 0.00 0.00 0.00 66.00 Month
2 9 1 23.22 76.53 -68.00 0.00 0.00 66.00 141.00 Month 3 10 0 48.20
155.17 -68.00 0.00 0.00 0.00 480.00 VAR/E Month 1 42 3 -23.52
167.46 -648.00 -67.00 0.00 69.00 211.00 Month 2 43 2 -10.63 110.07
-289.00 -72.00 0.00 1.00 361.00 Month 3 44 1 46.75 299.86 -687.00
0.00 0.00 69.00 1771.00 gE/E Month 1 42 3 -10.98 155.68 -266.00
-110.00 0.00 0.00 539.00 Month 2 43 2 -40.56 207.85 -1030.00 -72.00
0.00 0.00 367.00 Month 3 42 3 13.12 137.98 -226.00 -71.00 0.00
60.00 662.00 gEVAR/E Month 1 42 3 21.33 123.99 -425.00 -2.00 0.00
71.00 488.00 Month 2 43 2 29.33 179.40 -301.00 -66.00 0.00 68.00
831.00 Month 3 43 2 86.84 165.87 -136.00 0.00 0.00 199.00 709.00
CD8-TNF.alpha. Pool gE gE/Y Month 1 9 1 -22.33 33.50 -67.00 -67.00
0.00 0.00 0.00 Month 2 9 1 -7.44 40.26 -67.00 0.00 0.00 0.00 67.00
Month 3 9 1 0.56 59.77 -67.00 -1.00 0.00 0.00 140.00 gEVAR/Y Month
1 10 0 -34.00 65.20 -200.00 -67.00 0.00 0.00 0.00 Month 2 9 1
-30.22 68.11 -200.00 0.00 0.00 0.00 1.00 Month 3 10 0 -27.00 71.95
-200.00 -67.00 0.00 0.00 67.00 VAR/E Month 1 42 3 -6.71 90.36
-278.00 0.00 0.00 0.00 210.00 Month 2 42 3 -6.67 78.03 -219.00
-1.00 0.00 0.00 209.00 Month 3 44 1 -11.98 89.66 -347.00 0.00 0.00
0.00 206.00 gE/E Month 1 42 3 12.95 86.37 -184.00 0.00 0.00 7.00
421.00 Month 2 43 2 10.81 69.64 -220.00 0.00 0.00 65.00 220.00
Month 3 42 3 5.45 65.44 -140.00 0.00 0.00 0.00 305.00 gEVAR/E Month
1 42 3 14.10 72.50 -219.00 0.00 0.00 0.00 229.00 Month 2 43 2 6.28
57.99 -219.00 0.00 0.00 0.00 148.00 Month 3 43 2 39.09 129.10
-219.00 0.00 0.00 68.00 639.00 Varilrix gE/Y Month 1 9 1 76.33
232.77 -71.00 0.00 0.00 0.00 690.00 Month 2 9 1 468.78 1436.63
-143.00 0.00 0.00 0.00 4297.00 Month 3 9 1 -86.11 218.67 -632.00
-1.00 0.00 0.00 70.00 gEVAR/Y Month 1 10 0 -14.10 70.26 -141.00
-66.00 0.00 0.00 133.00 Month 2 9 1 29.89 146.91 -213.00 -66.00
0.00 69.00 273.00 Month 3 10 0 8.60 118.21 -213.00 -66.00 0.00
72.00 214.00 VAR/E Month 1 42 3 -40.52 354.84 -1873.00 -68.00 0.00
69.00 491.00 Month 2 43 2 -51.70 327.59 -1961.00 0.00 0.00 6.00
361.00 Month 3 44 1 110.00 653.00 -991.00 0.00 0.00 73.00 4087.00
gE/E Month 1 42 3 52.86 220.80 -455.00 -4.00 0.00 71.00 869.00
Month 2 43 2 17.14 356.23 -1493.00 -19.00 0.00 71.00 1312.00 Month
3 42 3 15.07 172.57 -571.00 0.00 0.00 70.00 449.00 gEVAR/E Month 1
42 3 15.48 312.96 -1711.00 0.00 0.00 134.00 694.00 Month 2 43 2
10.81 297.77 -1440.00 -66.00 0.00 67.00 838.00 Month 3 43 2 109.00
211.09 -148.00 0.00 0.00 205.00 904.00 gE/Y = gE-AS1/18-30 years;
gEVAR/Y = gE-AS1 + Varilrix/18-30 years VAR/E = Varilrix/50-70
years; gE/E = gE-AS1/50-70 years; gEVAR/E = gE-AS1 + Varilrix/50-70
years N = number of subjects with available results; N miss. =
number of subjects with missing results SD = Standard Deviation
Min, Max = Minimum, Maximum Q1, Q3 = First, Third quartile
TABLE-US-00020 TABLE C.4 Intracellular Cytokine Staining (ICS):
Inferential statistics: P- values from Kruskal-Wallis Tests for CD4
T cells at POST-PRE (Total Vaccinated Cohort) P_value at P_value at
P_value at Groups Month 1- Month 2- Month 3- T cells compared
Antigen Test PRE PRE PRE CD4 VAR\E and Pool gE ALL DOUBLES 0.0000
0.0004 0.0000 gEVAR\E CD40L 0.0000 0.0002 0.0000 IFN.gamma. 0.0000
0.0429 0.0000 IL2 0.0000 0.0000 0.0000 TNF.alpha. 0.0000 0.0009
0.0000 Varilrix ALL DOUBLES 0.0078 0.1736 0.0000 CD40L 0.0090
0.0727 0.0000 IFN.gamma. 0.0261 0.0447 0.0000 IL2 0.0067 0.0575
0.0000 TNF.alpha. 0.0620 0.1957 0.0000 VAR\E and Pool gE ALL
DOUBLES 0.0000 0.0004 0.0000 gE\E CD40L 0.0000 0.0001 0.0000
IFN.gamma. 0.0001 0.0880 0.0000 IL2 0.0000 0.0003 0.0000 TNF.alpha.
0.0009 0.0018 0.0000 Varilrix ALL DOUBLES 0.5370 0.1120 0.0000
CD40L 0.5137 0.2217 0.0000 IFN.gamma. 0.7205 0.2367 0.0000 IL2
0.5791 0.3599 0.0000 TNF.alpha. 0.8440 0.0880 0.0001 gE\E and Pool
gE ALL DOUBLES 0.3100 0.6612 0.3060 gEVAR\E CD40L 0.3996 0.7134
0.3350 IFN.gamma. 0.2366 0.7134 0.6835 IL2 0.4707 0.3629 0.4148
TNF.alpha. 0.3923 0.7134 0.2480 Varilrix ALL DOUBLES 0.1034 0.0049
0.1231 CD40L 0.1262 0.0054 0.1305 IFN.gamma. 0.0832 0.0021 0.0910
IL2 0.0921 0.0040 0.0831 TNF.alpha. 0.1179 0.0035 0.1372 VAR/E =
Varilrix/50-70 years gE/E = gE-AS1/50-70 years gEVAR/E = gE-AS1 +
Varilrix/50-70 years
TABLE-US-00021 SUPPLEMENTARY TABLE C.4 Intracellular Cytokine
Staining (ICS): Inferential statistics: P-values from
Kruskal-Wallis Tests for CD8 T cells at POST-PRE (Total Vaccinated
Cohort) Groups P_value at P_value at P_value at T cells compared
Antigen Test description Month 1-PRE Month 2-PRE Month 3-PRE CD8
VAR\E and Pool gE ALL DOUBLES 0.0575 0.2069 0.1364 gEVAR\E CD40L
0.1647 0.0113 0.1579 IFN.gamma. 0.1411 0.8759 0.0360 IL2 0.0456
0.1080 0.1442 TNF.alpha. 0.2938 0.3356 0.0499 Varilrix ALL DOUBLES
0.6363 0.8116 0.1785 CD40L 0.6944 0.4151 0.9266 IFN.gamma. 0.5953
0.8108 0.0486 IL2 0.6656 0.5567 0.1544 TNF.alpha. 0.3677 0.8788
0.2679 VAR\E and Pool gE ALL DOUBLES 0.2913 0.1159 0.9259 gE\E
CD40L 0.5885 0.2542 0.9217 IFN.gamma. 0.1900 0.3113 0.4158 IL2
0.1687 0.1288 0.8127 TNF.alpha. 0.3700 0.2008 0.8454 Varilrix ALL
DOUBLES 0.9067 0.9436 0.4197 CD40L 0.1382 0.4574 0.7783 IFN.gamma.
0.7445 0.6841 0.8567 IL2 0.1893 0.3980 0.3536 TNF.alpha. 0.6716
0.8132 0.6206 gE\E and Pool gE ALL DOUBLES 0.3308 0.6165 0.1380
gEVAR\E CD40L 0.4801 0.2231 0.1503 IFN.gamma. 0.9259 0.2911 0.1157
IL2 0.4306 1.0000 0.0678 TNF.alpha. 0.9797 0.6343 0.0646 Varilrix
ALL DOUBLES 0.5447 0.7670 0.0227 CD40L 0.2490 0.9913 0.8265
IFN.gamma. 0.4940 0.5395 0.0801 IL2 0.0705 0.1827 0.0264 TNF.alpha.
0.6200 0.7754 0.1098 VAR/E = Varilrix/50-70 years gE/E =
gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70 years
Lymphoproliferation Tables
TABLE-US-00022 [0176] LIST OF TABLES PAGE Table L.1 Descriptive
statistics on the Stimulating Index for lymphoproliferation (ATP
cohort for immunogenicity) . . . Table L.2 Lymphoproliferation:
Geometric Mean (ATP cohort for immunogenicity) . . . Table L.3
Lymphoproliferation: Inferential statistics on Stimultaing Index
(ATP cohort for immunogenicity) . . . Table L.4
Lymphoproliferation: Fold increase in Geometric Mean (GMR) (ATP
cohort for immunogenicity) . . .
TABLE-US-00023 TABLE L.1 Descriptive statistics on the Stimulating
Index for lymphoproliferation (ATP cohort for immunogenicity)
Stimulating N Concentration Antigen Group Timing N miss. Mean SD
Min Q1 Median Q3 Max 0.1 CPAU/ml VZV VAR/E Day 0 44 1 27.40 35.81
1.06 9.90 17.22 33.96 221.53 Month 1 44 1 29.82 25.70 0.94 9.36
23.66 45.66 92.16 Month 2 43 2 27.50 17.97 5.61 13.90 24.01 34.62
95.32 Month 3 43 2 29.34 24.53 6.72 13.62 20.55 36.93 124.46 gE/E
Day 0 43 2 21.86 17.72 1.21 9.66 17.76 28.15 91.05 Month 1 42 3
28.36 23.99 2.55 12.13 21.85 31.99 107.18 Month 2 42 3 27.47 22.90
1.62 14.85 20.71 32.88 138.04 Month 3 42 3 28.52 17.80 2.16 15.28
24.92 40.87 87.26 gE/Y Day 0 10 0 37.50 16.31 9.36 28.85 33.88
43.11 67.30 Month 1 9 1 42.87 28.45 5.42 24.89 38.04 44.23 102.82
Month 2 10 0 47.78 29.73 12.35 24.79 42.91 63.27 108.21 Month 3 10
0 47.77 25.64 8.87 25.98 44.79 74.27 82.49 gEVAR/E Day 0 42 2 19.30
21.14 1.35 6.27 17.79 22.49 136.88 Month 1 42 2 28.63 18.39 3.31
15.08 21.69 44.13 68.92 Month 2 42 2 30.92 24.15 1.97 11.97 23.06
49.92 96.33 Month 3 43 1 37.51 29.04 4.45 19.92 29.98 43.02 142.50
gEVAR/Y Day 0 10 0 49.64 22.42 29.45 39.22 41.28 57.67 106.72 Month
1 10 0 54.43 29.44 18.42 27.53 49.96 77.50 105.66 Month 2 9 1 51.83
24.74 11.07 34.87 52.85 74.34 82.69 Month 3 10 0 47.69 21.37 16.19
34.27 50.79 66.25 77.75 1 CPAU/ml VZV VAR/E Day 0 44 1 35.73 29.28
0.81 15.47 27.32 49.16 143.45 Month 1 44 1 43.08 28.02 1.58 22.67
34.43 58.49 122.15 Month 2 43 2 50.16 87.00 12.27 23.16 34.68 41.08
586.25 Month 3 43 2 36.02 23.73 8.56 17.36 28.63 46.93 116.31 gE/E
Day 0 43 2 32.54 26.56 4.59 17.91 27.88 39.18 139.71 Month 1 42 3
35.02 25.59 1.54 19.97 28.32 37.30 110.93 Month 2 42 3 35.73 22.52
1.46 21.91 31.20 41.19 107.68 Month 3 42 3 37.34 19.84 8.37 20.36
32.55 52.92 85.04 gE/Y Day 0 10 0 43.97 23.26 9.30 31.05 43.43
53.02 87.99 Month 1 9 1 51.10 35.47 6.31 37.60 44.42 57.47 132.58
Month 2 10 0 54.24 20.58 16.73 44.20 51.53 71.02 88.07 Month 3 10 0
56.04 37.19 13.72 35.84 45.31 68.27 143.80 gEVAR/E Day 0 42 2 29.13
24.91 1.20 12.54 23.70 39.25 146.46 Month 1 42 2 39.10 23.60 3.01
22.28 33.18 52.91 108.05 Month 2 42 2 41.11 26.67 7.67 21.90 35.00
55.75 126.34 Month 3 43 1 46.67 33.81 3.98 24.34 35.11 64.97 151.51
gEVAR/Y Day 0 10 0 59.60 32.75 27.66 39.90 46.26 75.47 138.10 Month
1 10 0 71.90 45.92 21.95 24.54 61.95 117.28 145.08 Month 2 9 1
68.71 36.16 28.77 34.89 69.01 94.82 128.29 Month 3 10 0 67.50 34.90
18.29 36.67 62.03 92.64 118.24 20 .mu.g/ml gE VAR/E Day 0 44 1 2.85
3.08 0.77 1.24 1.68 2.81 14.30 Month 1 44 1 3.53 3.87 0.57 1.34
2.03 4.31 20.96 Month 2 43 2 4.09 5.08 0.86 1.54 2.12 4.64 30.33
Month 3 43 2 3.66 3.01 0.60 1.53 2.48 5.72 14.05 gE/E Day 0 43 2
3.45 5.78 0.78 1.11 1.93 2.99 37.21 Month 1 42 3 12.69 10.72 0.76
5.41 8.88 17.63 45.95 Month 2 42 3 11.76 11.43 0.93 4.35 8.44 14.46
51.08 Month 3 42 3 30.34 21.83 1.97 15.69 21.16 40.12 101.46 gE/Y
Day 0 10 0 2.62 1.24 1.01 1.75 2.18 3.71 4.88 Month 1 9 1 19.09
15.69 2.17 6.67 13.12 35.81 43.15 Month 2 10 0 25.50 18.51 9.37
10.31 16.67 32.61 60.58 Month 3 10 0 37.46 21.69 4.72 27.78 31.11
52.84 76.56 gEVAR/E Day 0 42 2 3.84 7.70 0.59 1.17 1.49 3.61 49.50
Month 1 42 2 9.78 9.70 0.94 3.82 6.44 12.74 49.37 Month 2 42 2 9.89
8.26 0.79 3.88 8.90 13.30 43.68 Month 3 43 1 34.03 25.88 3.01 14.92
30.69 45.24 117.38 gEVAR/Y Day 0 10 0 5.78 5.63 1.85 2.42 3.26 7.53
20.39 Month 1 10 0 19.80 16.75 5.89 9.80 13.77 18.38 51.54 Month 2
9 1 24.38 14.38 8.00 9.03 22.40 38.49 44.11 Month 3 10 0 33.17
19.70 6.72 11.15 32.50 52.45 58.00 4 .mu.g/ml gE VAR/E Day 0 44 1
2.19 2.64 0.67 1.14 1.37 2.30 14.80 Month 1 44 1 3.47 3.55 0.26
1.31 1.87 4.70 17.20 Month 2 43 2 2.86 2.94 0.62 1.10 1.52 3.55
15.40 Month 3 43 2 3.30 2.69 0.47 1.36 2.36 4.20 11.36 gE/E Day 0
43 2 2.74 4.00 0.29 1.04 1.37 3.24 25.55 Month 1 42 3 10.08 12.66
0.65 2.84 4.49 14.49 67.46 Month 2 42 3 8.07 8.23 0.87 2.88 4.02
10.16 38.39 Month 3 42 3 28.13 21.36 1.78 14.58 20.83 36.28 104.34
gE/Y Day 0 10 0 2.85 1.95 0.85 1.12 2.70 3.69 6.21 Month 1 9 1
14.39 15.26 0.83 5.17 9.06 13.73 49.06 Month 2 10 0 19.81 14.60
4.71 9.75 13.64 28.99 51.20 Month 3 10 0 32.53 17.61 3.93 26.84
32.60 43.71 56.99 gEVAR/E Day 0 42 2 3.30 6.69 0.66 1.11 1.56 2.64
43.49 Month 1 42 2 7.40 9.69 0.88 2.33 4.80 8.10 58.76 Month 2 42 2
6.94 6.77 0.91 2.25 5.37 9.28 40.34 Month 3 43 1 30.45 25.08 4.02
9.63 24.42 43.85 110.76 gEVAR/Y Day 0 10 0 4.15 3.07 1.31 1.91 2.85
5.65 10.99 Month 1 10 0 15.96 10.92 3.86 10.96 13.00 15.98 40.77
Month 2 9 1 21.57 14.61 3.08 8.91 22.20 26.82 47.30 Month 3 10 0
30.11 19.58 3.58 8.61 30.13 51.66 54.23 gE/Y = gE-AS1/18-30 years
gEVAR/Y = gE-AS1 + Varilrix/18-30 years VAR/E = Varilrix/50-70
years gE/E = gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70
years N = number of subjects with available results N miss. =
number of subjects with missing results SD = Standard Deviation
Min, Max = Minimum, Maximum Q1, Q3 = First, Third quartile
TABLE-US-00024 TABLE L.2 Lymphoproliferation: Geometric Mean of
stimulation index (ATP cohort for immunogenicity) Stimulating N
Concentration Antigen Group Timing N miss. GMT Min Max 0.1 CPAU/ml
VZV VAR/E Day 0 44 1 15.69 1.06 221.53 Month 1 44 1 19.10 0.94
92.16 Month 2 43 2 22.62 5.61 95.32 Month 3 43 2 22.77 6.72 124.46
gE/E Day 0 43 2 15.38 1.21 91.05 Month 1 42 3 20.48 2.55 107.18
Month 2 42 3 21.23 1.62 138.04 Month 3 42 3 22.60 2.16 87.26 gE/Y
Day 0 10 0 33.69 9.36 67.30 Month 1 9 1 33.79 5.42 102.82 Month 2
10 0 39.66 12.35 108.21 Month 3 10 0 40.08 8.87 82.49 gEVAR/E Day 0
42 2 12.94 1.35 136.88 Month 1 42 2 22.67 3.31 68.92 Month 2 42 2
22.03 1.97 96.33 Month 3 43 1 28.62 4.45 142.50 gEVAR/Y Day 0 10 0
46.28 29.45 106.72 Month 1 10 0 46.68 18.42 105.66 Month 2 9 1
44.87 11.07 82.69 Month 3 10 0 42.48 16.19 77.75 1 CPAU/ml VZV
VAR/E Day 0 44 1 22.89 0.81 143.45 Month 1 44 1 33.48 1.58 122.15
Month 2 43 2 34.44 12.27 586.25 Month 3 43 2 29.76 8.56 116.31 gE/E
Day 0 43 2 24.61 4.59 139.71 Month 1 42 3 27.63 1.54 110.93 Month 2
42 3 29.72 1.46 107.68 Month 3 42 3 31.84 8.37 85.04 gE/Y Day 0 10
0 37.53 9.30 87.99 Month 1 9 1 39.83 6.31 132.58 Month 2 10 0 49.89
16.73 88.07 Month 3 10 0 46.60 13.72 143.80 gEVAR/E Day 0 42 2
20.59 1.20 146.46 Month 1 42 2 32.04 3.01 108.05 Month 2 42 2 33.52
7.67 126.34 Month 3 43 1 35.94 3.98 151.51 gEVAR/Y Day 0 10 0 53.44
27.66 138.10 Month 1 10 0 57.76 21.95 145.08 Month 2 9 1 60.28
28.77 128.29 Month 3 10 0 58.44 18.29 118.24 20 .mu.g/ml gE VAR/E
Day 0 44 1 2.05 0.77 14.30 Month 1 44 1 2.41 0.57 20.96 Month 2 43
2 2.75 0.86 30.33 Month 3 43 2 2.71 0.60 14.05 gE/E Day 0 43 2 2.15
0.78 37.21 Month 1 42 3 8.48 0.76 45.95 Month 2 42 3 7.76 0.93
51.08 Month 3 42 3 23.31 1.97 101.46 gE/Y Day 0 10 0 2.36 1.01 4.88
Month 1 9 1 12.71 2.17 43.15 Month 2 10 0 20.50 9.37 60.58 Month 3
10 0 30.14 4.72 76.56 gEVAR/E Day 0 42 2 2.08 0.59 49.50 Month 1 42
2 6.68 0.94 49.37 Month 2 42 2 7.09 0.79 43.68 Month 3 43 1 25.27
3.01 117.38 gEVAR/Y Day 0 10 0 4.29 1.85 20.39 Month 1 10 0 15.21
5.89 51.54 Month 2 9 1 20.06 8.00 44.11 Month 3 10 0 26.11 6.72
58.00 4 .mu.g/ml gE VAR/E Day 0 44 1 1.63 0.67 14.80 Month 1 44 1
2.37 0.26 17.20 Month 2 43 2 2.03 0.62 15.40 Month 3 43 2 2.48 0.47
11.36 gE/E Day 0 43 2 1.75 0.29 25.55 Month 1 42 3 5.74 0.65 67.46
Month 2 42 3 5.36 0.87 38.39 Month 3 42 3 20.95 1.78 104.34 gE/Y
Day 0 10 0 2.26 0.85 6.21 Month 1 9 1 8.55 0.83 49.06 Month 2 10 0
15.55 4.71 51.20 Month 3 10 0 25.27 3.93 56.99 gEVAR/E Day 0 42 2
1.94 0.66 43.49 Month 1 42 2 4.64 0.88 58.76 Month 2 42 2 4.90 0.91
40.34 Month 3 43 1 21.47 4.02 110.76 gEVAR/Y Day 0 10 0 3.31 1.31
10.99 Month 1 10 0 13.14 3.86 40.77 Month 2 9 1 16.14 3.08 47.30
Month 3 10 0 22.09 3.58 54.23 gE/Y = gE-AS1/18-30 years gEVAR/Y =
gE-AS1 + Varilrix/18-30 years VAR/E = Varilrix/50-70 years gE/E =
gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70 years N =
number of subjects with available results N miss. = number of
subjects with missing results GMT = Geometric Mean Titer LL, UL =
Lower, Upper Limit of 95% confidence interval Min, Max = Minimum,
Maximum
TABLE-US-00025 TABLE L.3 Lymphoproliferation: Inferential
statistics on Stimulating Index (ATP cohort for immunogenicity)
P-value Stimulating Kruskal- Groups compared Concentration Antigen
Timing Wallis gE\Y and 0.1 CPAU/ml VZV Day 0 0.1509 gEVAR\Y Month 1
0.3272 Month 2 0.6242 Month 3 0.8206 1 CPAU/ml VZV Day 0 0.3643
Month 1 0.3691 Month 2 0.5676 Month 3 0.3643 20 .mu.g/ml gE Day 0
0.0963 Month 1 0.7440 Month 2 0.8065 Month 3 0.7624 4 .mu.g/ml gE
Day 0 0.2899 Month 1 0.3691 Month 2 0.9349 Month 3 0.7624 VAR\E and
gE\E 0.1 CPAU/ml VZV Day 0 0.9594 Month 1 0.9037 Month 2 0.7317
Month 3 0.6226 1 CPAU/ml VZV Day 0 0.6899 Month 1 0.1062 Month 2
0.5041 Month 3 0.4657 20 .mu.g/ml gE Day 0 0.9526 Month 1 0.0000
Month 2 0.0000 Month 3 0.0000 4 .mu.g/ml gE Day 0 0.7342 Month 1
0.0002 Month 2 0.0000 Month 3 0.0000 gE\E and 0.1 CPAU/ml VzAg Day
0 0.3794 gEVAR\E Month 1 0.5489 Month 2 0.6939 Month 3 0.1903 1
CPAU/ml VzAg Day 0 0.5097 Month 1 0.2412 Month 2 0.3855 Month 3
0.3653 20 .mu.g/ml gE Day 0 0.6226 Month 1 0.1375 Month 2 0.6164
Month 3 0.5737 4 .mu.g/ml gE Day 0 0.6226 Month 1 0.4524 Month 2
0.8161 Month 3 0.9160 VAR\E and 0.1 CPAU/ml VzAg Day 0 0.5059
gEVAR\E Month 1 0.5922 Month 2 0.8812 Month 3 0.0913 1 CPAU/ml VzAg
Day 0 0.2688 Month 1 0.5803 Month 2 0.7450 Month 3 0.1253 20
.mu.g/ml gE Day 0 0.7690 Month 1 0.0000 Month 2 0.0000 Month 3
0.0000 4 .mu.g/ml gE Day 0 0.3553 Month 1 0.0016 Month 2 0.0000
Month 3 0.0000 gE/Y = gE-AS1/18-30 years gEVAR/Y = gE-AS1 +
Varilrix/18-30 years VAR/E = Varilrix/50-70 years gE/E =
gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70 years
TABLE-US-00026 TABLE L.4 Lymphoproliferation: Fold increase in
Geometric Mean (GMR) (ATP cohort for immunogenicity) Stimulating
Ratio N Concentration Antigen Group POST/PRE N miss. GMR LL UL Min
Max 0.1 CPAU/ml VZV VAR/E Month 1/Month 0 43 2 1.54 1.01 2.34 0.04
27.05 Month 2/Month 0 42 3 1.65 1.05 2.58 0.09 52.56 Month 3/Month
0 42 3 2.04 1.39 2.99 0.29 48.92 gE/E Month 1/Month 0 41 4 1.39
1.01 1.92 0.15 18.32 Month 2/Month 0 41 4 1.42 0.94 2.12 0.02 20.08
Month 3/Month 0 41 4 1.86 1.38 2.52 0.13 14.83 gE/Y Month 1/Month 0
9 1 0.68 0.28 1.61 0.05 1.53 Month 2/Month 0 10 0 1.01 0.73 1.40
0.44 1.72 Month 3/Month 0 10 0 1.16 0.88 1.53 0.61 2.17 gEVAR/E
Month 1/Month 0 40 4 2.10 1.45 3.04 0.13 31.67 Month 2/Month 0 40 4
2.06 1.36 3.11 0.06 58.19 Month 3/Month 0 41 3 3.22 2.28 4.54 0.86
82.33 gEVAR/Y Month 1/Month 0 10 0 0.87 0.63 1.20 0.34 1.44 Month
2/Month 0 9 1 0.87 0.52 1.43 0.18 1.37 Month 3/Month 0 10 0 0.64
0.37 1.11 0.12 1.06 1 CPAU/ml VZV VAR/E Month 1/Month 0 43 2 1.73
1.17 2.56 0.06 86.17 Month 2/Month 0 42 3 1.63 1.06 2.51 0.08
124.71 Month 3/Month 0 42 3 1.79 1.24 2.59 0.47 70.57 gE/E Month
1/Month 0 41 4 1.18 0.90 1.53 0.05 13.44 Month 2/Month 0 41 4 1.16
0.82 1.64 0.01 18.94 Month 3/Month 0 41 4 1.64 1.23 2.18 0.09 19.91
gE/Y Month 1/Month 0 9 1 0.75 0.32 1.73 0.05 1.38 Month 2/Month 0
10 0 1.14 0.91 1.42 0.78 1.81 Month 3/Month 0 10 0 1.21 1.01 1.46
0.83 1.69 gEVAR/E Month 1/Month 0 40 4 1.85 1.40 2.43 0.46 15.65
Month 2/Month 0 40 4 1.82 1.33 2.49 0.13 48.63 Month 3/Month 0 41 3
2.52 1.88 3.38 0.88 80.53 gEVAR/Y Month 1/Month 0 10 0 0.93 0.75
1.17 0.48 1.33 Month 2/Month 0 9 1 0.98 0.79 1.22 0.60 1.26 Month
3/Month 0 10 0 0.76 0.46 1.26 0.15 1.29 20 .mu.g/ml gE VAR/E Month
1/Month 0 43 2 1.35 1.04 1.75 0.16 11.02 Month 2/Month 0 42 3 1.46
1.10 1.94 0.27 20.90 Month 3/Month 0 42 3 1.90 1.42 2.54 0.39 26.66
gE/E Month 1/Month 0 41 4 4.26 2.98 6.07 0.22 74.67 Month 2/Month 0
41 4 3.62 2.41 5.46 0.05 28.79 Month 3/Month 0 41 4 13.90 9.16
21.07 0.31 205.73 gE/Y Month 1/Month 0 9 1 3.84 1.17 12.65 0.11
12.75 Month 2/Month 0 10 0 7.45 5.15 10.77 3.91 15.23 Month 3/Month
0 10 0 12.50 7.41 21.10 3.14 37.77 gEVAR/E Month 1/Month 0 40 4
3.89 2.90 5.22 0.95 28.69 Month 2/Month 0 40 4 3.86 2.54 5.87 0.03
38.27 Month 3/Month 0 41 3 17.92 12.65 25.38 1.38 172.56 gEVAR/Y
Month 1/Month 0 10 0 3.07 1.40 6.72 0.38 15.77 Month 2/Month 0 9 1
4.09 2.10 7.98 1.85 15.42 Month 3/Month 0 10 0 4.25 1.60 11.30 0.28
15.60 4 .mu.g/ml gE VAR/E Month 1/Month 0 43 2 1.75 1.36 2.25 0.08
8.82 Month 2/Month 0 42 3 1.40 1.03 1.89 0.12 36.27 Month 3/Month 0
42 3 2.17 1.60 2.95 0.51 66.05 gE/E Month 1/Month 0 41 4 3.32 2.34
4.69 0.31 36.29 Month 2/Month 0 41 4 3.10 2.02 4.76 0.04 25.22
Month 3/Month 0 41 4 14.32 9.39 21.82 0.26 110.97 gE/Y Month
1/Month 0 9 1 2.88 0.75 11.06 0.07 21.03 Month 2/Month 0 10 0 5.91
3.32 10.52 2.49 31.74 Month 3/Month 0 10 0 10.95 5.62 21.37 3.07
48.19 gEVAR/E Month 1/Month 0 40 4 2.90 2.26 3.71 0.54 22.40 Month
2/Month 0 40 4 2.96 2.08 4.21 0.10 35.98 Month 3/Month 0 41 3 16.40
11.62 23.15 1.51 237.09 gEVAR/Y Month 1/Month 0 10 0 3.43 1.61 7.32
0.41 16.60 Month 2/Month 0 9 1 3.98 1.76 9.01 0.91 31.37 Month
3/Month 0 10 0 4.65 1.72 12.60 0.30 19.48 gE/Y = gE-AS1/18-30 years
gEVAR/Y = gE-AS1 + Varilrix/18-30 years VAR/E = Varilrix/50-70
years gE/E = gE-AS1/50-70 years gEVAR/E = gE-AS1 + Varilrix/50-70
years N = number of subjects with available results N miss. =
number of subjects with missing results GMR = Geometric Mean ratio
LL, UL = Lower, Upper Limit of 95% confidence interval for GMR Min,
Max = Minimum, Maximum
CONCLUSIONS
[0177] The gE AS1 vaccine and the concomitant delivery of gE AS1
with the OKA strain both provoke a good immune response in
comparison to the response obtained by the OKA strain alone.
Sequence CWU 1
1
11546PRTVaricella zoster 1Met Gly Thr Val Asn Lys Pro Val Val Gly
Val Leu Met Gly Phe Gly1 5 10 15Ile Ile Thr Gly Thr Leu Arg Ile Thr
Asn Pro Val Arg Ala Ser Val 20 25 30Leu Arg Tyr Asp Asp Phe His Ile
Asp Glu Asp Lys Leu Asp Thr Asn 35 40 45Ser Val Tyr Glu Pro Tyr Tyr
His Ser Asp His Ala Glu Ser Ser Trp 50 55 60Val Asn Arg Gly Glu Ser
Ser Arg Lys Ala Tyr Asp His Asn Ser Pro65 70 75 80Tyr Ile Trp Pro
Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His 85 90 95Glu His His
Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu 100 105 110Arg
Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp 115 120
125Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe
Lys Gly145 150 155 160Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu
Ile Glu Val Ser Val 165 170 175Glu Glu Asn His Pro Phe Thr Leu Arg
Ala Pro Ile Gln Arg Ile Tyr 180 185 190Gly Val Arg Tyr Thr Glu Thr
Trp Ser Phe Leu Pro Ser Leu Thr Cys 195 200 205Thr Gly Asp Ala Ala
Pro Ala Ile Gln His Ile Cys Leu Lys His Thr 210 215 220Thr Cys Phe
Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr225 230 235
240Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu
Phe Asp 260 265 270Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly
Val Leu Lys Val 275 280 285Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val
Tyr Ile Trp Asn Met Arg 290 295 300Gly Ser Asp Gly Thr Ser Thr Tyr
Ala Thr Phe Leu Val Thr Trp Lys305 310 315 320Gly Asp Glu Lys Thr
Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro 325 330 335Arg Gly Ala
Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser 340 345 350Val
Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His 355 360
365Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr
His Pro385 390 395 400Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser
Gly Cys Thr Phe Thr 405 410 415Ser Pro His Leu Ala Gln Arg Val Ala
Ser Thr Val Tyr Gln Asn Cys 420 425 430Glu His Ala Asp Asn Tyr Thr
Ala Tyr Cys Leu Gly Ile Ser His Met 435 440 445Glu Pro Ser Phe Gly
Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys 450 455 460Phe Val Asp
Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val465 470 475
480Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro
Pro Thr 500 505 510Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu
Ile Thr Pro Val 515 520 525Asn Pro Gly Thr Ser Pro Leu Ile Arg Tyr
Ala Ala Trp Thr Gly Gly 530 535 540Leu Ala545
* * * * *