Figure 5. VAD affected T cell functionality in the periphery in viral infection.

A) Naïve CD44⁻CD4⁺ and B) CD44⁻CD8⁺ T cells were purified from spleens of naïve mice. Cells were labeled with CFSE and cultured in vitro for 4 days with the anti-CD3/CD28 antibody stimulation. At last 5 h before harvest, PMA/Ionomycin and Brefeldin A were added into the culture system. Cell proliferation and intracellular IFN-γ expression were measured by flow cytometry. C) Splenocytes were isolated from VAC and VAD mice, followed by adoptively transferring into CD45.1 transgenic mice. D) Splenocytes were isolated from naïve CD45.1 transgenic mice, followed by adoptively transferring into VAC and VAD mice. These recipient mice were infected with LCMV and sacrificed at 6 dpi. The adoptive transferred cells were gated first, and the percentages of cytokine-producing T cells were analyzed by flow cytometry. The data are shown as mean ± SEM of three mice per group from a single representative experiment. The experiment was repeated three times independently. A two-tailed Student’s t-test was used to compare the two groups. ** P<0.01, NS, no significance.