Figure 3.

Vaginal LCMV infection elicits delayed and dampened expansion of virus-specific effector CD8 T cells leading to delayed viral control in LFRT. (A–C and H–L) Naive CD45.1⁺ P14 CD8 T cells (P14 cells) were isolated, and 500,000 (days 0.5–4) or 20,000 (day 5 onward) P14 cells were adoptively transferred into CD45.2⁺ WT recipient mice 1 d before i.p. or i.vag. infection with 2 × 10⁵ PFU LCMV. (A–C) Viral copies in total RNA from the indicated tissues of infected WT mice were determined by qRT-PCR. (D–G) WT, CD8^−/−, MHC-II^−/−, or RAG1^−/− mice were infected i.vag., and LCMV copies at the indicated times and in the indicated tissues were determined as in A–C. (H) LFRT tissues from mice in A–C were digested, and extracted immune cells were analyzed by flow cytometry. Absolute counts of P14 cells at days 4 and 5 are shown. (I) Representative flow cytometry plots showing the abundance of CD45.1⁺ P14 cells in total cells isolated from the LFRT at days 5 and 7 after infection from P14 immune chimeras in A–C. (J) Longitudinal fraction of P14s in the PBMCs of i.p. or i.vag. infected animals. (K–L) Total number of P14s in indicated tissues at the indicated time points after i.p. or i.vag. infection. Data represent one of two (A–G and J) or three (H, I, K, and L, days 7–9) independent experiments. n = 3–4 (A–C), 10 (WT in D), 3 (CD8^−/− in D; F; and d4 in H), 4 (MHC^−/− in D; d5 in H; K; and L), 7 (i.p. in J), or 5 (i.vag. in J) mice per group. Error bars represent mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001, unpaired Student’s t test. UI, uninfected.