Fig. 1

IL-10-mediated inhibition of IFNγ secretion by human CD4⁺ T cells requires dendritic cells. 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 72 h, 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 < 0.01, ***P < 0.001 using one-way ANOVA (e) or unpaired Student’s t test (a, g, h)