FIG. 2.
Humoral and cellular anti-gD responses elicited after prime-boosting with rVSV-gD. Mice were primed intranasally with the indicated dose in PFU of rVSVCh-gD or rVSVCh-Gag at week 0, boosted intranasally with the same dose of rVSVI-gD or rVSVI-Gag at week 4, and euthanized to collect sera and spleen cells at week 8. (A) Individual serum samples were tested for presence of IgG1 and IgG2a anti-gD antibodies by ELISA and for anti-HSV-2 neutralizing antibodies. Spleen cells were pooled per treatment group and were restimulated in vitro with UV-inactivated HSV-2 to evaluate the cellular immune responses. (B) Three-day antigen-restimulated pooled spleen cell populations were evaluated by fluorescence-activated cell sorting for internal IFN-γ expression. (C) Culture supernatants obtained from 3-day restimulated pooled spleen cells were analyzed by cytokine bead array testing for the presence of TNF-α, IFN-γ, IL-2, IL-4, and IL-5. Concentrations of IL-4 were below background and are not shown. (D) Five-day antigen-restimulated pooled spleen cell populations were examined for their ability to recognize and lyse HSV-2-infected and mock-infected target cells.