Comparison of Heterologous Prime-Boost Strategies against Human Immunodeficiency Virus Type 1 Gag Using Negative Stranded RNA Viruses
Figure 7
Recall response in mice after NDV-Gag prime and heterologous boost.
(A) Schedule of prime-boost-challenge vaccinations. Mice were primed intramuscularly on Day 0 and 6. 35 days later the mice were boosted. Mice were challenged intraperitoneally 33 days post prime. PBS mice were included as a negative control. 5 days after challenge, mice were euthanized and spleens harvested for analysis of the cellular response. (B–D) Each point is representative of splenocytes from one mouse (n = 5 per group). (B) The quantity of activated HIV-1 Gag-specific CD8+ T cells in the spleen was analyzed by flow cytometry and the percentage of cells are shown. Activated cells were determined by gating on CD62Llo cells and HIV-1 Gag-specific cells were determined by tetramer staining against the H2d restricted AMQMKLETI epitope. (C–E) The functionality of the CD8+ T cells was measured by intracellular cytokine staining for (C) IFNγ+, (D) TNFα+, and (E) IFNγ+TNFα+ cells after stimulation of the cells with AMQMKLETI peptide. Statistical analysis was performed using one-WAY ANOVA. All comparisons to PBS are statistically significant. Results shown are presented as the mean. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.