Fig. 1: The differential activation and expansion of human CD4+ T cell subsets following exposure to bacteria derived sAgs. | Nature Communications

Fig. 1: The differential activation and expansion of human CD4+ T cell subsets following exposure to bacteria derived sAgs.

From: Staphylococcal superantigens evoke temporary and reversible T cell anergy, but fail to block the development of a bacterium specific cellular immune response

Fig. 1

250,000 PBMCs were cultured for 48 h either alone (Nil) or with 1% v/v of bacterial supernatants harvested from overnight cultures of the following strains, JKD6159, JKD6159::sea, JKD6159::seb, Newman, Newmanâ–³sea and the percentages of CD4+ T cells expressing the activation marker CD69 was measured by flow cytometry. a Representative FACs profiles gated on CD4+ T cells showing the percentage of CD69+ cells. b The percentage of CD69+CD4+ T cells. Symbols represent individual donors, bars represent the mean ± SEM. Data pooled from 3 donors (one way ANOVA, Dunnett’s multiple comparison). c–g 20,000 moDCs were cultured (1:1) with matched CFSE labelled bulk PBMCs for 7 days either alone (Nil) or with 0.1%, 1.0% or 10% v/v of bacterial supernatants harvested from overnight cultures of the following strains, JKD6159, JKD6159::sea, JKD6159::seb, Newman, Newmanâ–³sea. The absolute number of divided (CFSElo) CD4+ T cells was measured by flow cytometry. Graphs depict the percentages of maximum CD4+ T cell division per experiment (lines represent individual donors, Two-way ANOVA, Dunnett’s multiple comparison). h The absolute number of IFNγ and TNFα producing CD4+ T cells, measured by intracellular cytokine staining, following a brief in vitro restimulation with bacterial supernatant. Symbols represent individual donors, bars represent the mean ± SEM. Data pooled from 3 donors (log transformation, Two-way ANOVA, Dunnett’s multiple comparison). i Representative flow cytometry profiles gated on HLA-DR+ cells and CD4+ T cells, which were further subdivided into naïve (CD27+CD45RA+), Tcm (CD27+CD45RA−) and Tem (CD27−CD45RA−). j–k 20,000 HLA-DR+ cells and CFSE labelled CD4+ Tcm, Tem and Tn subsets sort purified from matched donor PBMCs were cultured 1:1 for 7 days either alone, (Nil) or with 0.1% (v/v) of bacterial supernatants harvested from overnight cultures of the following strains, JKD6159, JKD6159::sea, JKD6159::seb, Newman, Newmanâ–³sea, COL or COLâ–³sebâ–³sekâ–³seq. j The absolute number of divided (CFSElo) CD4+ T cells and (k) the proportion producing TNFα (TNFα+) following a brief in vitro restimulation. Symbols represent individual donors, bars represent the mean ± SEM. Data pooled from 4 donors (two way ANOVA, Tukey’s multiple comparison). *p < 0.5, **p < 0.01, ***p < 0.001, and ****p < 0.0001.

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