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. 2016 Nov 9;20(5):584–595. doi: 10.1016/j.chom.2016.10.007

Figure 6.

Figure 6

SteD Suppresses T Cell Proliferation

(A) Mouse BMDCs were infected with the indicated bacterial strains and total MHCII surface levels (I-A/I-E haplotypes) were quantified by flow cytometry at 20 hr post uptake.

(B) BMDCs were infected as in (A), and B7.2 surface levels were quantified by flow cytometry at 20 hr post uptake.

(A and B) Surface levels of proteins in infected cells are represented as a percentage of those in uninfected cells from the same sample and are the means ± SD from four independent experiments.

(C) Infected BMDCs were incubated with OVA peptide and co-cultured with T cells labeled with CFSE for 3 days. T cell proliferation was analyzed by flow cytometry after labeling cells with anti-CD3, anti-V alpha2, and anti-CD4 antibodies. The uninfected BMDCs incubated or not with OVA-peptide were used as controls. The results shown represent the % of T cells that proliferated and are the means ± SD from quadruplicate samples in three independent experiments.

(D) Representative FACS histograms showing T cell proliferation measured by CFSE levels in different conditions.

(E) Dendritic cells from mesenteric lymph nodes of mice infected with WT-GFP or ΔsteD-GFP Salmonella by oral gavage were isolated at 48 hr post inoculation and total MHCII surface levels were measured by flow cytometry. The data represent surface levels of MHCII in infected BMDCs as a percentage of those in uninfected cells from the same sample and are the means ± SD from three independent experiments.

(F) Activated T cells (CD25+CD44+ and CD62L−CD44+) as a percentage of total CD4+ T cells isolated from the spleens of mice infected by Salmonella WT or ΔsteD strains at day 17 post inoculation. The data in (A) and (B) were analyzed by comparison with WT and in (C) by comparison with uninfected +OVA by one-way ANOVA followed by Dunnett’s multiple comparison test. The data in (E) were analyzed by Student’s t test. The data in (F) were analyzed by two-tailed pairwise t test using pairs shown in Table S2. ∗∗∗∗p < 0.0001, ∗∗p < 0.01, ∗p < 0.02, and not significant, ns.