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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: J Immunol. 2016 Aug 29;197(7):2748–2761. doi: 10.4049/jimmunol.1501926

Figure 8. Effect of Anti-Ad5 antibodies in the immunogenicity of Ad5 and Ad5/3.

Figure 8

(A) Neutralization of Ad5 (Closed Bars) or Ad5/3 (Open Bars) by sera of mice immunized with Ad5. Viruses at an MOI of 10 were incubated with 5μl of sera from Ad5 immunized CB6F1/J mice before infecting HeLa cells plated in 96-well plates. The infectivity was determined by measurement of the luciferase transgene. The results present the reciprocal mean neutralization titers able to neutralize 50% of the luciferase activity ** p<0.01 by two tailed unpaired Student’s T test. The experiments were done in triplicate. (B) Specific SYVPSAEQI/APC Tet+ CD8+ T cells induced by immunization with Ad5/33PP or Ad5PP regimens in mice (n=5 per group) previously immunized with empty Ad5 to induce neutralizing antibodies. Tetramer recognition was assessed 10 days after the final protein boost and is expressed as arithmetic mean values ± standard deviations (SD). **p<0.01 by Kruskal-Walllis with Dunns’ post-test comparsions are presented between the immunization groups and the control group. (C) Anti-PyLPC/RMC antibody titers measured 20 days after the third and final immunization. Endpoint ELISA titers at 0.5 optical density were determined using the recombinant protein PyLPC/RMC and serum samples from mice immunized with the different heterologous regimens and expressed as arithmetic mean values ± standard deviations (SD). (D) Frequencies of CD4+ and (E) CD8+ IFN-γ, IL-2 or TNF-α secreting T cells after stimulation with PyLPC/RMC. Spleens were obtained from previously Ad5 sensitized mice, after immunization with the heterologous regimens. Bars represent mean responses ± standard deviations for five mice per group.