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. Author manuscript; available in PMC: 2018 Dec 6.
Published in final edited form as: Sci Transl Med. 2010 Oct 27;2(55):55ra78. doi: 10.1126/scitranslmed.3000448

Fig. 3.

Fig. 3.

ICOS is critical for the expansion of human TH17 cells. (A and B) The frequency (A) and absolute number (B) of CCR4+CCR6+CD4+ T cells over time were assessed by flow cytometry from peripheral blood CD4+ T cells cultured in TH17-polarizing conditions and activated with antibodies to CD3/CD28 or CD3/ICOS beads. (C) CD27 and CD62L expression was measured on day 10 on these cells with flow cytometry. (D) On the days indicated, CD28- or ICOS-engaged TH17-polarized CD4+ T cells were stimulated with PMA-ionomycin and the frequency of cells secreting IL-17A and IFN-γ was assessed via flow cytometry. (E) The frequency of CD28- or ICOS-engaged TH17-polarized cells coproducing IL-17A and/or IFN-γ was determined at the end of their primary expansion (ranging from days 9 to 14) in several different normal donors (n = 8). (F and G) RORC2 and T-bet expression in these treated cells was measured with RT-PCR on days 3 and 10. Representative of two to three experiments.