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. Author manuscript; available in PMC: 2020 Feb 15.
Published in final edited form as: Mucosal Immunol. 2019 Aug 15;12(5):1201–1211. doi: 10.1038/s41385-019-0194-9

Figure 1. IL-10-mediated inhibition of IFNγ secretion by human CD4+ T cells requires dendritic cells.

Figure 1.

(a, b) Peripheral blood mononuclear cells or (c, d) CD4+ T cells from an IL10RA-deficient patient and adult healthy controls (n=4) were stimulated with soluble anti-CD3 or anti-CD3/CD28 beads (bead-to-cell ratio 1:2) with or without IL-10. After 24 h or 48 h, supernatants were assayed for IFNγ using an ELISA. (e) Purified CD14+ monocytes, immature moDCs, LPS-matured moDCs, CD4+ T cells and anti-CD3/CD28-activated CD4+ T cells (48 h) were analyzed by flow cytometry for expression of IL-10RA and IL-10RB chains. Delta Mean fluorescence intensity (MFI) values (MFI-minus-MFI of the isotype control) are shown for n=4–8 adult healthy individuals per group. (f) Control whole blood or purified CD4+ T cells were stimulated with IL-10 for 60 min followed by quantification of STAT3 phosphorylation (Tyr705) by flow cytometry. (g) Allogeneic MLRs were performed using LPS-matured moDCs and CD4+ T cells from an IL10RA-deficient patient or healthy individuals. The bacterial superantigen Staphylococcal enterotoxin B (SEB) was added to all conditions in the presence or absence of IL-10. After 72h, supernatants were assayed for IFNγ using an ELISA. (h) Allogeneic MLRs were performed using moDCs and CD4+ T cells from healthy individuals or AD-HIES patients carrying STAT3 mutations. SEB was added to all conditions in the presence or absence of LPS and IL-10. After 72 h, supernatants were assayed for IFNγ using a Cytometric Bead Array. Results are mean ± SD (B, D, H) or mean ± SEM (A, C E, G) of a representative of at least two independent experiments. *P<0.05, ***P<0.001 using one-way ANOVA (E) or unpaired Student’s t test (A, G, H)