by Peter Abbink, Rafael A. Larocca, Rafael A

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Protective efficacy of multiple vaccine platforms against Zika virus challenge in rhesus monkeys by Peter Abbink, Rafael A. Larocca, Rafael A. De La Barrera, Christine A. Bricault, Edward T. Moseley, Michael Boyd, Marinela Kirilova, Zhenfeng Li, David Ng’ang’a, Ovini Nanayakkara, Ramya Nityanandam, Noe B. Mercado, Erica N. Borducchi, Arshi Agarwal, Amanda L. Brinkman, Crystal Cabral, Abishek Chandrashekar, Patricia B. Giglio, David Jetton, Jessica Jimenez, Benjamin C. Lee, Shanell Mojta, Katherine Molloy, Mayuri Shetty, George H. Neubauer, Kathryn E. Stephenson, Jean Pierre S. Peron, Paolo M. de A. Zanotto, Johnathan Misamore, Brad Finneyfrock, Mark G. Lewis, Galit Alter, Kayvon Modjarrad, Richard G. Jarman, Kenneth H. Eckels, Nelson L. Michael, Stephen J. Thomas, and Dan H. Barouch Science Volume ():aah6157 August 4, 2016 Published by AAAS

Fig. 1 Immunogenicity of the ZIKV PIV vaccine. Immunogenicity of the ZIKV PIV vaccine. (A) Env-specific ELISA titers and (B) ZIKV-specific microneutralization (MN50) titers following immunization of rhesus monkeys by the s.c route with 5 μg ZIKV PIV vaccine at weeks 0 and 4 (red arrows). The maximum measurable log MN50 titer in this assay was 3.86. Cellular immune responses by IFN-γ ELISPOT assays to prM, Env, Cap, and NS1 at (C) week 2 and (D) week 6. Red bars reflect medians. Peter Abbink et al. Science 2016;science.aah6157 Published by AAAS

Fig. 2 Protective efficacy of the ZIKV PIV vaccine. Protective efficacy of the ZIKV PIV vaccine. PIV vaccinated and sham control rhesus monkeys (n = 8/group) were challenged by the s.c route with 106 VP (103 PFU) ZIKV-BR or ZIKV-PR. Each group contained 6 female and 2 male animals. Viral loads are shown in (A) plasma, (B) urine, (C) CSF, (D) colorectal secretions, and (E) cervicovaginal secretions. Viral loads were determined on days 0, 1, 2, 3, 4, 5, 6, 7 for the plasma samples (A) and on days 0, 3, 7 for the other samples [(B) to (E)]. Data are shown for all 8 animals in each panel, except for the 6 females for cervicovaginal secretions in (E). P value reflects Fisher’s exact test. Peter Abbink et al. Science 2016;science.aah6157 Published by AAAS

Fig. 3 Adoptive transfer studies in mice. Adoptive transfer studies in mice. (A) Env-specific serum ELISA titers and (B) ZIKV-specific microneutralization (MN50) titers in serum from recipient Balb/c mice (n = 5/group) 1 hour following adoptive transfer of 5-fold serial dilutions (Groups I, II, III, IV) of IgG purified from PIV vaccinated rhesus monkeys or sham controls. (C) Plasma viral loads in mice following challenge with 105 VP (102 PFU) ZIKV-BR. (D and E) Immune correlates of protection. Red bars reflect medians. P values reflect t tests. Peter Abbink et al. Science 2016;science.aah6157 Published by AAAS

Fig. 4 Adoptive transfer studies in rhesus monkeys. Adoptive transfer studies in rhesus monkeys. (A) ZIKV-specific microneutralization (MN50) titers in serum from recipient rhesus monkeys (n = 2/group) 1 hour following adoptive transfer of 5-fold dilutions (Groups I, II) of IgG purified from PIV vaccinated rhesus monkeys or sham controls. (B) Plasma viral loads in rhesus monkeys following challenge with 106 VP (103 PFU) ZIKV-BR. Red bars reflect medians. Peter Abbink et al. Science 2016;science.aah6157 Published by AAAS

Fig. 5 Immunogenicity of the ZIKV DNA-prM-Env and RhAd52-prM-Env vaccines. Immunogenicity of the ZIKV DNA-prM-Env and RhAd52-prM-Env vaccines. (A) ZIKV-specific microneutralization (MN50) titers following immunization of rhesus monkeys by the i.m. route with 5 mg DNA-prM-Env vaccine at weeks 0 and 4 (red arrows) or a single immunization with 1011 vp RhAd52-prM-Env at week 0. (B) Cellular immune responses by IFN-γ ELISPOT assays to prM, Env, Cap, and NS1 at week 6 for the DNA vaccine or at week 4 for the RhAd52 vaccine. Red bars reflect medians. Peter Abbink et al. Science 2016;science.aah6157 Published by AAAS

Fig. 6 Protective efficacy of the ZIKV DNA-prM-Env and RhAd52-prM-Env vaccines. Protective efficacy of the ZIKV DNA-prM-Env and RhAd52-prM-Env vaccines. DNA vaccinated, RhAd52 vaccinated, and sham control rhesus monkeys (n = 4/group) were challenged by the s.c route with 106 VP (103 PFU) ZIKV-BR or ZIKV-PR. Plasma viral loads are shown. Peter Abbink et al. Science 2016;science.aah6157 Published by AAAS