DNA and Protein Vaccination Confers Protection Upon Mucosal Challenge with Heterologous SIVsmE660 (OA 10.04)
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1 DNA and Protein Vaccination Confers Protection Upon Mucosal Challenge with Heterologous SIVsmE660 (OA 10.04) Rashmi Jalah Human Retrovirus Pathogenesis Section, Vaccine Branch, CCR, National Cancer Institute at Frederick, Frederick, Maryland, USA AIDS Vaccine 2011 Meeting Bangkok, k Thailand Wednesday, September 14th, 2011
2 Vaccination with DNA as the ONLY Vaccine Modality Leads to: Induction of long-lasting (~2 years) humoral and cellular immune responses Ptl Patel, et al., Vaccine 2010 Dissemination of antigen-specific T cells and Ab to mucosal sites Patel, et al., Vaccine 2010 Protection from high viremia upon SIVmac251 high dose challenge Rosati, et al., JVirol, 2005; Rosati, et al., PNAS 2009
3 Objectives Improve humoral immune responses by combining i DNA vaccine with protein Study vaccine efficacy upon repeated lowdose heterologous SIVsmE660 mucosal challenge
4 Vaccine: Optimized SIV Plasmid DNAs Alone or with Protein (Co-delivered, or as Prime-Boost) Group (N=8) Vaccine Vaccination 1: 0 Vaccination 2: month 2 Vaccination 3: month 4 Vaccination 4: month 9 1 DNA Only DNA DNA DNA DNA 2 3 DNA + Protein codelivered DNA prime- Protein Boost DNA + AT-2 SIV DNA + AT-2 SIV DNA + AT-2 SIV DNA + AT-2 SIV DNA DNA AT-2 SIV AT-2 SIV 4 Sham-Vaccinated Sham DNA Sham DNA Sham DNA Sham DNA SIVmac239 DNA mixture: DNAs expressing SIV immunogens (Env, Gag, Pol, Nef, Tat, Vif) coinjected with optimized rhesus IL-12 DNA as molecular adjuvant Intramuscular (IM) delivery of DNA by in vivo electroporation (Inovio) Protein: AT-2 inactivated SIVmac239 particles
5 Presence of High Levels of Vaccine-induced % SIV-spe cific IFNγ+ T ce ells SIV-specific IFN-γ + T cells (up to 3.5%) DNA DNA+Protein DNA prime Only coimmunization Protein Boost EP2w2 EP4w2 DOC EP2w2 EP4w2 DOC EP2w2 EP4w2 DOC Vaccine:D D D+P D+P D P
6 Immunization with SIVmac239 Based Vaccines Induces Humoral Immune Responses ses Against the Heterologous SIVsmE660 Env Binding Abs to E660 env Avidity to E660 env Neutralizing Abs to E660 (CG_7G) p<0.05 p= p= DNA DNA +protein DNA prime protein boost DNA DNA +protein DNA prime protein boost DNA DNA +protein DNA prime protein boost 7 months after last vaccination (Day of Challenge)
7 Inclusion of DNA in Vaccine Protocol Promotes the Maintenance of Binding Antibody Levels Overtime EP4w2 7 months DOC (7 months after EP4) Log bindin ng antibody titers to S IVmac239 P < DNA DNA DNA prime +protein protein boost ~4x drop ~3x drop P < P< DNA DNA +protein DNA prime protein boost ~14x drop (Least persistent) Co-immunization with DNA+Protein confers highest and longest lasting Env binding antibodies
8 Heterologous Challenge with SIVsmE660 4 vaccinations (SIVmac239) EP1 EP2 EP3 EP4 Repeated Low Dose Mucosal Challenge with heterologous SIVsmE660 (83% aa identity in env) 14 weekly exposures Month months 10 months of follow up
9 Virus Acquisition Upon SIVsmE660 Challenge nfected % unin 2/8 = 25% (DNA+protein co immunization) remain uninfected No of SIVsmE660 exposures Trend for slower virus acquisition in the vaccine groups 2 animals in DNA+proteincoimm coimmunization group remained uninfected Group size (n=8) does not provide the power to find differences among individual groups
10 DNA/protein Vaccination Delays Virus Acquisition % unin nfected Combination of our controls and a cohort of Indian Rhesus Macaques infected with SIVsmE660 (R. Amara & H. Robinson) using: the same virus lot (V. Hirsch) the same amount of virus the same route of infection No of SIVsmE660 exposures Slower virus acquisition (p=0.05) in vaccinees (N=24) compared to controls (N=23) Cochran-Mantel-Haensel test, stratified for TRIM5α
11 Vaccinated Macaques Show Significant Lower Median Peak Viremia After Challenge with SIVsmE660 Decrease of peak viremia i by 17l 1.7 log p= Median peak log SIV RNA copies/m ml Vaccinees N=22 Controls N=8 Significant virological benefit also after stratification of animals according to TRIM5a genotype
12 Potent Long-lasting Protection from SIVsmE660 Challenge in 75% of Vaccinees (18 of 24 animals) Monitored for 40 weeks Sterilizing immunity: 2 Elite controller: 10 Viremic Controller: 6 s load Virus Weeks of infection Chronic Median VL (wk 6-40): <threshold animals did not control challenge (chronic median VL of 5.59)
13 Correlation of E660 Env Binding Ab and Avidity with Slower Acquisition of the SIVsmE660 Challenge Virus Log bind ing ab to E660 at DOC Not infected R= p = Avidity to E660 at DOC Not infected R= p = No of SIVsmE660 exposures
14 Inverse Correlation of SIV-specific Cellular Effector Memory Responses with Control of Viremia peak viru us load R= R= p = p = Log % IFN+ CD4+ EM T cells %IFN+GrzB+ CD4+ EM T cells (Square root) (Square root) Data are from EP2wk2; Correlations are also found at EP4w2
15 Conclusions DNA+protein codelivery improves immunogenicity High and persistent levels of cellular immune responses Increase of magnitude, longevity and quality of humoral responses to Env higher avidity babs and higher neutralizing Abs Heterologous challenge outcome: Significant delay in virus acquisition Significant reduction in VL and long-lasting control (>40 weeks) of viremia 75% of the vaccinees control viremia after challenge with SIVsmE660 Provides sterilizing immunity in 2 macaques (vaccinated with DNA+protein) Correlation of E660 binding Abs and avidity with slower virus acquisition No correlation of SIVmac239 binding Abs and avidity with acquisition Inverse correlation of cytotoxic CD4 effector memory with peak VL
16 Acknowledgements Vanessa Hirsch (NIAID) Viraj Kulkarni Yongjun Guan, IHV, Baltimore Candido Alicea Brandon Keele, Julian Bess & Jeff Lifson Jenifer Bear (AIDS and Cancer Virus Program, SAIC, Rachel Kelly Beach Frederick) Barbara K. Felber, NCI Frederick David Venzon (NCI) David Montefiori (Duke) Rama Amara (Emory) & Harriet Vainav Patel Robinson (Geovax) Antonio Valentin Margherita Rosati Welkin Johnson (Harvard Med School) Agneta von Gegerfelt KE K.E. Broderick k& N. Y. Sardesai i(inovio George N. Pavlakis, NCI Frederick Pharmaceuticals) Phil Markham + ABL staff (Kensington)
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