Figure 45.

Flow cytometric analysis of IL-10 production by human CD4⁺ T-cell subsets. (A) Conventional human CD4⁺ T-cells (Complete gating strategy see Chapter human CD4⁺T-cells) were isolated according to IL-7R and CCR6 expression and stimulated for 4 or 30 hours with anti-CD3 Abs in the absence or presence of anti-CD28 Abs or 100 U/ml IL-2. The production of IL-2 and IL-10 is shown. The frequencies of IL-10⁺ cells are reported (Statistics: paired student’s t-test, n=5). (B and C) Representative intracellular IL-10 and IFN-γ stainings of FACS-purified CD4⁺CXCR5⁺ICOS⁺ tonsillar TFH-cells (B) or of human blood CD4⁺IL-7R^lowCD25⁻T-cells (C) stimulated with PMA and Ionomycin for 4 h. (D) Same as in C, but IL- 10 was analyzed with a secretion assay and combined with CD40L surface staining. (E) IL-10 secretion of human CD4⁺T-cells following overnight stimulation of total PBMC with SEB gated on CD4⁺LAG3⁺CD49b⁺Tr1- cells. 2010 Häringer et al. Originally published in J. Exp. Med. https://doi.org/10.1084/jem.20091021 [163] (Fig. 45A) and https://doi.org/10.1084/jem.20082238 [165] (Fig. 45C).