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. Author manuscript; available in PMC: 2019 May 12.
Published in final edited form as: Nat Microbiol. 2018 Nov 12;4(1):62–70. doi: 10.1038/s41564-018-0280-x

Fig. 3. Depletion of MRC-1 abolishes pneumolysin induced cytokine inhibition and enhances T cell activation.

Fig. 3

(a) Uptake of T4R and T4R∆ply by WT and MRC-1 siRNA treated DCs (N=3 donors). The uptake was expressed as a percentage relative to untreated DCs. Data represent mean ± S.E.M. **** denotes P<0.0001 by two-way ANOVA with Bonferroni post-test. (b) Wild type DCs (control) or MRC-1 siRNA treated DCs were infected with T4R and secretion of IL-12, TNF-α and IL-6 was measured in culture supernatants (N=3 donors). Data represent mean ± S.E.M. **** denotes P<0.0001 and ** denotes P<0.01 by two-way ANOVA with Bonferroni post-test. (c-d) Wild type or MRC-1 siRNA treated DCs were infected with T4R, T4R∆ply or recombinant PLY (rPLY) (200 ng/mL) for 24 hrs and co-cultured with naïve CD4+ T cells for 5 days. Secretion of (c) IFN-γ and (d) IL-4 was measured in culture supernatants (N=5 donors). Data represent mean ± S.E.M. **** denotes P<0.0001, ** denotes P<0.01, * denotes P<0.05 by two-way ANOVA with Bonferroni post-test. (e) FoxP3 expression in human naïve CD4+ T cells upon co-culture with DCs (control or MRC-1 siRNA treated) infected with T4R or T4R∆ply. Data are representative of three independent experiments. (f) Percentage FoxP3+ CD4+ T cells upon co-culture with murine BMDMs (from wild type or MRC-1-/- mice) that were infected with heat-killed strain D39 or mutant derivatives lacking capsule (D39∆cps), PLY (D39∆ply) or a double mutant (D39ΔcpsΔply) or purified PLY. Data represent mean ± S.E.M. N=3, two way-ANOVA with Bonferroni multiple comparison test. * P < 0.05; **P< 0.01.