Figure 4
Primary immunization combined with boost vaccination results in an increase in the number of germinal center and isotype-switched B cells and IFN-γ-secreting, HSV-1 gB-specific CD8+ T cells in 0∆NLS vaccinated mice. C57BL/6 mice (n = 5–8/group) were injected in the footpad with the HSV-1 parental (GFP105) or live-attenuated HSV-1 0ΔNLS at 105 plaque forming units (PFU) in 10 µl PBS. At day 21 post immunization, the mice were boosted with the same amount of either parental or live-attenuated HSV-1 virus and left for 30 days. Thirty days after boosting, mice were exsanguinated to evaluate (A) germinal center (GC) B cell (CD19+GL-7+CD95+) and (B) isotype-switched B cell (CD19+IgM-IgD-) formation along with (C) analysis of the number of pre-activated B cells secreting IgG1 after polyclonal stimulation, and (D) HSV-1 neutralization titer from sera obtained from mice 30 days post-booster. (E) The spleens were removed and processed to single-cell suspensions in which 1 × 106 cells/well were stimulated overnight with gB peptide, SSIEFARL (10 µg/ml) to measure IFN-γ secreting CD8+ T cells by ELISPOT assay. Bars represent the mean ± SEM, **p < .01, *p < .05 comparing the indicated groups as determined by Student’s t-test.
