Figure 3. TCR Tg CD4 T cells mediate protection and demonstrate a recall response following challenge.
Rag1−/− mice (4–5 per group) were mock treated or intravenously injected with indicated numbers of CD4 T cells isolated from the spleens of naïve C57BL/6J wild type or Tg mice, and intravaginally inoculated with CM001 5 days later. Survival was monitored daily, and an exact log rank test was used to analyze survival differences between polyclonal and Tg groups (** P < 0.01). (B) Recall response of CD4+Vβ10+ Tg CD4 T cells to secondary infection. Following primary infection, Tg mice were treated with doxycycline, rested for 9 weeks, and re-challenged with CM001. On day 7 post-challenge, genital tract CD4 T cells were harvested, stimulated with 5 μg/ml C. muridarum, and analyzed for IFNγ and TNF production by intracellular cytokine staining. (C) Indicated numbers of naïve Tg or polyclonal CD4 T cells were adoptively transferred to Tcra−/− mice 5 days prior to intravaginal infection with CM001, and the course of primary infection was monitored by culture of lower genital tract swabs. Significance was determined by two-way RM ANOVA. Data represent the mean ± SEM of 10 mice per group. Comparison of individual days for 103 Tg versus 106 polyclonal: * P < 0.05, ** P < 0.01. Comparison of primary infection course between groups: P=NS for 106 Tg versus 103 Tg. *** P =0.0001 for 103 Tg versus 106 Polyclonal. **** P <0.0001 for all remaining group comparisons. (D) Immune mice were treated with doxycycline on days 52–56 post-infection, rested for 5 weeks, re-challenged with CM001, and infection monitored by culture of vaginal swabs. Significance was determined by two-way RM ANOVA. Data represent the mean + SD of 4–5 mice per group. Comparison of groups over primary infection course: P=NS for 103 Tg versus 106 polyclonal, 106 Tg versus 103 or 106 polyclonal. *P < 0.05 for 103 Tg versus 106 Tg. **** P <0.0001 for all remaining group comparisons.