Figure 8 | Mucosal Immunology

Figure 8

From: Hypertrophy of infected Peyer’s patches arises from global, interferon-receptor, and CD69-independent shutdown of lymphocyte egress

Figure 8

CD69-dependent and -independent lymphocyte retention mechanisms. (a) Quantitative RT-PCR analysis of sphingosine-1-phosphate receptor 1 (S1PR1) expression in fluorescence-activated cell sorting (FACS)-sorted CD19+CD62L+ cells from non-infected and day 7-infected Peyer’s patches (PPs). A total of 13 (d0) and 12 (d7) samples generated in eight independent sorts. For one sample, PPs were pooled from one to two mice. Mean+s.d. of samples. (b) CXCR4 expression on CD19+ cells in PPs of non-infected (ctrl), LPS-treated (LPS), and day 7-infected (d7) mice, determined using flow cytometry. Two independent experiments, total four to six mice per group. Symbols represent PP pools from individual mice. (c) Selective retention of activated CD4+ T cells. CD69wtOTII or CD69−/−OTII cells labeled with cell proliferation dye were transferred to congenic recipients. Fifty microgram OVA were given po, PPs were fluorescein isothiocyanate (FITC)-injected on day 3 and analyzed on day 4. Symbols represent individual PPs. One-way analysis of variance (ANOVA), Tukey post test. FACS plot from CD69wtOTII transfer shows transferred CD4+ cells. Transferred CD4+ cells were subgated on division cycles. Mean+s.d. of individual PPs. Two-way ANOVA, Bonferroni post test comparing genotypes. Four experiments with total 4 (no OVA), 11 (CD69wtOTII+OVA), 8 (CD69−/−OTII+OVA), and three mice per group (LPS). (d) Retention of non-activated OTII cells. OTII cells were transferred to congenic recipients. Mice were infected or not infected and analyzed on days 5–6, 1 day after FITC injection. Left FACS plot is gated on viable PP cells, subsequent plots gated on indicated subpopulations. Two experiments, total four (d0) and six (d5–6) mice. Symbols represent individual PPs. Mann–Whitney test. Fold retention infected/non-infected PPs. Mean+s.d. of experiments.

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