Figure 3.
S1PR5 expression alters CD8+ T cell trafficking in lymphoid organs. (A and B) Effector OT-I T cells carrying distinct congenic markers were transduced with control (Ctrl-RV) or S1PR5 (S1PR5-RV) GFP-expressing RVs and cotransferred into recipient mice. Transduced cells were isolated from the spleen and inguinal LNs 8 d after transfer. (A) Ratio of Ctrl-RV and S1PR5-RV cells isolated from indicated organs is shown. (B) Mice were administered i.v. with anti-CD45 antibody (i.v.+) to label vasculature-associated cells. Frequencies of i.v.+ Ctrl-RV and S1PR5-RV cells from indicated organs are shown. (C–E) Effector CD45.1+ gBT-I T cells were transduced with control or S1PR5 RVs and transferred into recipient mice. Spleens were harvested 7 d after transfer. (C) Confocal images of the spleen showing the localization of GFP+ Ctrl-RV or S1PR5-RV cells (green) in relation to B cells (B220+, blue) and endothelial cells (CD31+, red). Selected areas are magnified (right panels) to highlight the boundary between WP and RP. Lack of CD31 staining was used to delineate WP/RP separation (dotted lines). (D) Number of GFP+ cells on whole spleen sections were quantified. (E) Frequencies of Ctrl-RV and S1PR5-RV cells located in the WP and RP were calculated. n = 4 mice per group. (F) Effector OT-I T cells carrying distinct congenic markers were transduced with Ctrl-RV or S1PR5-RV, cotransferred into recipient mice, and isolated from the spleen and LNs 2 h later. Mice were i.v. labeled before harvest. Ratio of Ctrl-RV and S1PR5-RV cells isolated from the indicated organs were normalized to splenic i.v.+ cells. In A, B, and F, data are representative of two independent experiments, with n = 8–10 mice per experiment. Wilcoxon test was used in B, Mann–Whitney test in E, and one-way ANOVA in F. *, P < 0.05; **, P < 0.01; ****, P < 0.0001. Graph shows mean ± SD. TEFF, effector T cell.

S1PR5 expression alters CD8+ T cell trafficking in lymphoid organs. (A and B) Effector OT-I T cells carrying distinct congenic markers were transduced with control (Ctrl-RV) or S1PR5 (S1PR5-RV) GFP-expressing RVs and cotransferred into recipient mice. Transduced cells were isolated from the spleen and inguinal LNs 8 d after transfer. (A) Ratio of Ctrl-RV and S1PR5-RV cells isolated from indicated organs is shown. (B) Mice were administered i.v. with anti-CD45 antibody (i.v.+) to label vasculature-associated cells. Frequencies of i.v.+ Ctrl-RV and S1PR5-RV cells from indicated organs are shown. (C–E) Effector CD45.1+ gBT-I T cells were transduced with control or S1PR5 RVs and transferred into recipient mice. Spleens were harvested 7 d after transfer. (C) Confocal images of the spleen showing the localization of GFP+ Ctrl-RV or S1PR5-RV cells (green) in relation to B cells (B220+, blue) and endothelial cells (CD31+, red). Selected areas are magnified (right panels) to highlight the boundary between WP and RP. Lack of CD31 staining was used to delineate WP/RP separation (dotted lines). (D) Number of GFP+ cells on whole spleen sections were quantified. (E) Frequencies of Ctrl-RV and S1PR5-RV cells located in the WP and RP were calculated. n = 4 mice per group. (F) Effector OT-I T cells carrying distinct congenic markers were transduced with Ctrl-RV or S1PR5-RV, cotransferred into recipient mice, and isolated from the spleen and LNs 2 h later. Mice were i.v. labeled before harvest. Ratio of Ctrl-RV and S1PR5-RV cells isolated from the indicated organs were normalized to splenic i.v.+ cells. In A, B, and F, data are representative of two independent experiments, with n = 8–10 mice per experiment. Wilcoxon test was used in B, Mann–Whitney test in E, and one-way ANOVA in F. *, P < 0.05; **, P < 0.01; ****, P < 0.0001. Graph shows mean ± SD. TEFF, effector T cell.

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