Figure 4.
S1PR5 induces CD8+ T cell egress from nonlymphoid tissues. (A–C) Effector gBT-I T cells carrying distinct congenic markers were transduced with Ctrl-RV or S1PR5-RV and cotransferred intradermally (i.d.) into the flank of naive mice. (A) Ratios of Ctrl-RV and S1PR5-RV cells isolated from the indicated organs normalized to those in the spleen. (B) CD69 and CD103 expression on Ctrl-RV or S1PR5-RV cells isolated from the skin 8 d after transfer. (C) Skin was harvested 4 or 8 d after i.d. cell transfer and cultured overnight. Transduced cells were enumerated from the skin and culture supernatant (S/N) and shown as a ratio. (D–F) Effector gBT-I or OT-I T cells were transduced with control or S1PR5 RVs and cotransferred i.v. into naive mice whose flanks were treated with DNFB. (D) Mice were i.v. labeled before harvest. (D and E) Ratios of Ctrl-RV and S1PR5-RV cells were enumerated from the indicated organs and normalized to numbers obtained from the spleen at the indicated times after transfer. (F) CD69, CXCR6, and CD103 expression on Ctrl-RV (black) or S1PR5-RV (blue) cells isolated from the indicated organs 8 d after transfer. (G) P14 T cells were transduced with control or S1PR5 RVs and cotransferred i.v. into mice infected with LCMV and treated with DNFB. Transduced cells were isolated from the indicated tissues 8 d after transfer. (H) Effector CD8+ T cells from WT (CD45.1+CD45.2+) or Ccr7−/− (CD45.2+) were transduced with Ctrl-RV or S1PR5-RV and transferred i.d. into CD45.1 recipient mice. Shown is the ratio Ccr7−/− and WT cells from the skin 7 d after transfer. In A and B, data are representative of two independent experiments, with n = 6–8 mice per experiment. In C, data are pooled from two independent experiments, with n = 4 mice per time point. In D, data are representative of two independent experiments, with n = 6–8 mice per time point per experiment. In E–G, data are representative of two to four independent experiments, with n = 6–10 mice per experiment. In H, data are pooled from three independent experiments, with n = 4 mice per group per experiment. Unpaired t test was used in A, multiple Wilcoxon test in C, multiple Mann–Whitney test in D, Kruskal-Wallis test in E and G, and Mann–Whitney test in H. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001. SG, salivary gland; TEFF, effector T cell.

S1PR5 induces CD8+ T cell egress from nonlymphoid tissues. (A–C) Effector gBT-I T cells carrying distinct congenic markers were transduced with Ctrl-RV or S1PR5-RV and cotransferred intradermally (i.d.) into the flank of naive mice. (A) Ratios of Ctrl-RV and S1PR5-RV cells isolated from the indicated organs normalized to those in the spleen. (B) CD69 and CD103 expression on Ctrl-RV or S1PR5-RV cells isolated from the skin 8 d after transfer. (C) Skin was harvested 4 or 8 d after i.d. cell transfer and cultured overnight. Transduced cells were enumerated from the skin and culture supernatant (S/N) and shown as a ratio. (D–F) Effector gBT-I or OT-I T cells were transduced with control or S1PR5 RVs and cotransferred i.v. into naive mice whose flanks were treated with DNFB. (D) Mice were i.v. labeled before harvest. (D and E) Ratios of Ctrl-RV and S1PR5-RV cells were enumerated from the indicated organs and normalized to numbers obtained from the spleen at the indicated times after transfer. (F) CD69, CXCR6, and CD103 expression on Ctrl-RV (black) or S1PR5-RV (blue) cells isolated from the indicated organs 8 d after transfer. (G) P14 T cells were transduced with control or S1PR5 RVs and cotransferred i.v. into mice infected with LCMV and treated with DNFB. Transduced cells were isolated from the indicated tissues 8 d after transfer. (H) Effector CD8+ T cells from WT (CD45.1+CD45.2+) or Ccr7−/− (CD45.2+) were transduced with Ctrl-RV or S1PR5-RV and transferred i.d. into CD45.1 recipient mice. Shown is the ratio Ccr7−/− and WT cells from the skin 7 d after transfer. In A and B, data are representative of two independent experiments, with n = 6–8 mice per experiment. In C, data are pooled from two independent experiments, with n = 4 mice per time point. In D, data are representative of two independent experiments, with n = 6–8 mice per time point per experiment. In E–G, data are representative of two to four independent experiments, with n = 6–10 mice per experiment. In H, data are pooled from three independent experiments, with n = 4 mice per group per experiment. Unpaired t test was used in A, multiple Wilcoxon test in C, multiple Mann–Whitney test in D, Kruskal-Wallis test in E and G, and Mann–Whitney test in H. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001. SG, salivary gland; TEFF, effector T cell.

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