FIGURE 4.

Immune responses of HF10-immunized mice. (A) Serum was obtained from HF10-immunized mice (n = 3), and its neutralizing ability against HSV-1 strains (HF10 and KOS) and an HSV-2 strain (186) was investigated by the reduction in plaque formation. (B) Mice were immunized with UV-inactivated HF10 or HF10. After 4 weeks, serum (n = 3) collected and neutralizing ability against HF10 was assayed by the reduction in plaque formation. (C) IFN-γ produced by splenocytes stimulated with HSV-2 strain 186-infected NIH3T3 cells. Splenocytes (1 × 10⁷ cells/dish) from unimmunized (n = 3), UV-inactivated HF10-immunized (n = 3), or HF10-immunized (n = 3) mice were incubated with 186-infected NIH3T3 cells (1 × 10⁷ cells/dish) for 5 and 20 h, and the supernatants were collected. Supernatants from three mice were combined and assayed for IFN-γ concentrations using an ELISA. (D) IFN-γ levels in mice vagina after HSV-2 challenge. Genital tracts of unimmunized, UV-inactivated HF10-immunized, and HF10-immunized mice were washed 0, 1, and 3 days after challenge with strain 186, and the washes were assayed for IFN-γ concentrations by ELISA. *P < 0.05 between unimmunized mice and HF10-immunized mice. ND; not detected.