Quantifying HSV-specific T Cell Responses

HSV-specific T cell responses were analyzed as previously described42 (link). At day 6 post-infection, CD4⁺ and CD8⁺ T cells were isolated from the draining lymph nodes of mice infected intravaginally with 10⁶ pfu of TK- HSV-2 using anti-CD4- or anti-CD8-conjugated microbeads (Miltenyi Biotech) according to the manufacturer’s instructions. 1 × 10⁵ CD4⁺ and CD8⁺ T cells were co-cultured with 2 × 10⁵ splenocytes as antigen presenting cells and restimulated with various amounts of heat-inactivated HSV-2 or HSV glycoprotein B (gB) peptide (SSIEFARL) at 37 °C for 72 h. IFN-γ production in supernatants was measured by ELISA.