Novel HIV IL-4R antagonist vaccine strategy can induce
both high avidity CD8 and excellent B cell immunity
Charani Ranasinghe
Molecular Mucosal Vaccine Immunology Group, Dept Immunology
The John Curtin School of Medical Research
The Australian National University
 Mucosal vaccination induces high quality/avidity HIV-specific
CD8 T cells
Ranasinghe et al. J. Immunol 2007
 Induction of high quality/avidity HIV-specific CD8 T cells
following mucosal vaccination is associated with lower
expression of IL-4/IL-13 by CD8 T cells
 Absence of IL-4/IL-13 induces high avidity HIV-specific CD8 T
cells
Ranasinghe et al. Euro J Immunol 2009
Ranasinghe et al. Mucosal Immunology 2013
2
Construction of poxviral vector-based vaccines that co-express IL-4R
antagonist using homologous recombination
IL-4R antagonist
HIV gag/pol/env
Recombinant fowlpox virus (rFPV) - priming vaccine
IL-4R antagonist
HIV gag/pol
Recombinant vaccinia virus (rVV) or Modified Vaccinia Ankara (rMVA) - booster vaccine
intranasal/ intramuscular (i.n./i.m.)
combined mucosal/systemic strategy
Jackson Boyle Ranasinghe Methods in Molecular Biology (2014)
Novel IL-4R antagonist adjuvanted vaccine will bind to IL-4R and
transiently block IL-4/IL-13 signaling via the STAT6 pathway
IL-13
γc
IL-13Rα1
IL-4Rα
IL-4
IL-13Rα2m
12 hrs
STAT6
96 hrs
Ranasinghe et al Cytokine and Growth Factor Reviews (In press)
Jackson Worley Trivedi Ranasinghe (submitted)
4
i.n./i.m. IL-4R antagonist adjuvanted vaccine strategy
can induce HIV-specific CD8 T cells of high avidity
Inclusion of the inhibitor in the priming vaccination
is crucial to induce the high avidity T cell repertoire
Jackson Worley Trivedi Ranasinghe (submitted)
5
Inclusion of the inhibitor in the i.n. rFPV priming vaccination
is crucial to induce high avidity CD8 T cell repertoire
FPV HIV/VV HIV
FPV HIV/VV HIVIL-4C118
FPV HIVIL4C118/VVHIVIL-4C118
9.8%
21.8%
19.6%
1.16%
0.76%
0.98%
FPV HIVIL-4C118/VV
HIV
6.9%
Spleen
Genitorectal
nodes
Kd-Gag
0.38%
CD8
Jackson Worley Trivedi Ranasinghe (submitted)
Novel vaccines can enhance both systemic & mucosal HIV-specific
poly-functional HIV-specific CD8 T cell immunity
CD8+IFN-g+
CD8+TNF-a+
CD8+IFN-g+ TNF-a+
CD8+
Spleen
17.3%
Iliac nodes
1.0%
98.4%
78.1%
0.5%
3.5%
13.3%
Lung
2.5%
0.8%
0.5%
0.3%
32.1%
15.9%
78.9%
81.7%
59.7%
2.2%
96.7%
83.7%
0.1%
1.1%
Lung nodes
24.6%
15.4%
67.8%
5.2%
8.2%
Control Spl
4.7%
0.7%
94.1%
2.9%
Control illiac n.
Control lung
0.55%
99.4%
93.8%
0.03%
0.5%
5.58%
0.45%
7.6%
Contro lung n.
99.3%
0.02%
0.12%
0.01%
0.65%
0.01%
79.0%
12.5%
93.2%
5.1%
90.7%
7.3%
95.6%
2.9%
8.6%
1.9%
1.7%
Spleen
Iliac nodes
Ranasinghe Mucosal Immunology 2013
Lung
1.5%
Lung nodes
i.n./i.m. IL-4R antagonist adjuvanted HIV vaccine strategy
induces excellent CD8 T cell mediated protective immunity
* * *
P < 0.05 compared to
control vaccination
Ranasinghe et al Mucosal Immunology 2013;
Jackson Worley Trivedi Ranasinghe (submitted)
8
i.n./i.m. IL-4R antagonist adjuvanted vaccine strategy can
induce Gag-specific antibody class switching
6 weeks
12 weeks
* 0.0567
IgG1
* 0.0256
* 0.0256
* 0.0566
*** 0.0006
IgG2a
Jackson Worley Trivedi Ranasinghe (submitted)
Statistics were calculated using Mann – Whitney U test
9
i.n. delivery of FPV-HIV IL-4R antagonist adjuvanted vaccine recruits
unique antigen presenting cells to the lung mucosae responsible for
the induction of high avidity CD8 T and excellent B cell immunity
FPV-HIV - unadjuvanted
FPV-HIV IL-4RC118 (IL-4R antagonist)
73.5%
45.1%
CD11C+ CD11b+ CD103CD11b
1.92%
1.03%
CD103
Ranasinghe et al Mucosal Immunology 2013 ;
Trivedi Jackson Ranasinghe (Submitted)
10
Unique features of the novel i.n./i.m. HIV IL-4R antagonist
adjuvanted vaccine strategy.




Help recruit unique antigen presenting cell subsets to the lung
mucosae
Induce enhanced high quality/avidity mucosal & systemic HIV
Gag-specific CD8 T cell immunity*
HIV Gag-specific antibody class switching (IgG1 and IgG2a)*
Env-specific IgG1 following a second i.m. Env protein booster**
Induce triple action immunity
The immune responses induced are consistent with
• HIV controllers* and
• Features of partial protective efficacy in the RV144 trial**
=> Platform technology against other chronic pathogens
11
Acknowledgements
Molecular Mucosal Vaccine Immunology Group:
Ronald Jackson
Annette Buchanan
Lisa Pavlinovic, Megan Glidden, Sherry Tu
Students: Danushka Wijsundara, Shubhanshi
Trivedi, Matthew Worley
ANU/BRF - Kerong Zhang
Collaborators:
David Boyle - CSIRO AAHL
JCSMR/MCRF - Harpreet Vohra, Mick Devoy
ANU/Animal services staff
John Stambas - Deakin Uni/ CSIRO AAHL
Robert Center - Burnet Institute/ Uni of Melbourne
12