Fig. 3.

Tregs are required to mount a protective immune response at the site of infection subsequent to genital HSV-2 infection. Foxp3^WT or Foxp3^DTR mice were infected with HSV-2 and treated with DT. (A) For ex vivo cultures, CD4⁺ T cells were isolated from the dLNs of naïve or infected Foxp3^WT or Foxp3^DTR mice and cultured for 3 days with irradiated, heat-inactivated HSV-2-pulsed APCs. IFN-γ was detected by enzyme-linked immunosorbent assay (ELISA). (B) Mice were infected and treated with DT as in Fig. 2, dLNs were collected 2 days after infection, and extracts were prepared for IFN-α detection by ELISA. (C and D) IFN-γ present in vaginal washes collected at various times after infection (C) and IFN-α from vaginal washes collected 2 days after infection (D) were measured by ELISA. (E) At the indicated times after infection, dLNs and vaginal tracts were subjected to flow cytometric analysis. Naïve mice received DT according to the same schedule as infected mice.