T cell-mediated responses in genital tract (GT) following vaginal Chlamydia trachomatis (C.t.) challenge. Mice were immunized twice with a mixture of Hirep1, CTH93, and CAF01 with 2-week intervals using either the subcutaneous (SC) or simultaneous (SIM) immunization strategy. A CAF01 adjuvant control was included. Five weeks after the immunization protocol described in Figure , mice were challenged intravaginally with C.t., and on days 0, 1, 3, 6, 21, and 35 postinfection pools of cells from the GT, or iliac LNs of four mice were restimulated in vitro with a mixture of Hirep1 and CTH93. (A,C) Supernatants from restimulated iliac LNs were harvested and interferon gamma (IFNγ) and interleukin (IL)-17 levels determined by enzyme-linked immunosorbent assay. (B,D) Frequencies of IFNγ and IL-17-producing antigen-specific T cells (CD4+CD44high) were assessed by intracellular flow cytometry.