(A–C) Depo-provera treated WT C57BL/6 mice were intravaginally infected with 20,000 pfu of HSV-1 bbd or the rescue mutant bbdR, and were analyzed on days six and eight post infection. Total number of lymphocytes (A), and CD4+ T-cells (B) from the iliac and lingual draining lymph nodes is depicted. (C) Isolated CD4+ T-cells were co-cultured with naïve splenocytes in the presence of heat-inactivated HSV-1. IFN-γ secretion from CD4+ T cells was measured by ELISA. (D–G) CD4−/−, CD8−/−, and WT control C57BL/6 mice were intravaginally infected with 2,000 pfu of the indicated strain of HSV-1. Vaginal washes of CD4−/− (D) and CD8−/− (F) mice were collected daily and viral titers were measured in Vero cells. CD4−/− (E) and CD8−/− (G) mice were monitored for survival every 24 hours post infection. Error bars represent SEM of n > 5 mice per group. Data are pooled from four (A–C) or three (D–F) independent experiments. See also Figure S5.