IL-21 and TGF-beta are required for differentiation of human T(H)17 cells

Abstract

The recent discovery of CD4(+) T cells characterized by secretion of interleukin (IL)-17 (T(H)17 cells) and the naturally occurring regulatory FOXP3(+) CD4 T cell (nT(reg)) has had a major impact on our understanding of immune processes not readily explained by the T(H)1/T(H)2 paradigm. T(H)17 and nT(reg) cells have been implicated in the pathogenesis of human autoimmune diseases, including multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease and psoriasis. Our recent data and the work of others demonstrated that transforming growth factor-beta (TGF-beta) and IL-6 are responsible for the differentiation of naive mouse T cells into T(H)17 cells, and it has been proposed that IL-23 may have a critical role in stabilization of the T(H)17 phenotype. A second pathway has been discovered in which a combination of TGF-beta and IL-21 is capable of inducing differentiation of mouse T(H)17 cells in the absence of IL-6 (refs 6-8). However, TGF-beta and IL-6 are not capable of differentiating human T(H)17 cells and it has been suggested that TGF-beta may in fact suppress the generation of human T(H)17 cells. Instead, it has been recently shown that the cytokines IL-1beta, IL-6 and IL-23 are capable of driving IL-17 secretion in short-term CD4(+) T cell lines isolated from human peripheral blood, although the factors required for differentiation of naive human CD4 to T(H)17 cells are still unknown. Here we confirm that whereas IL-1beta and IL-6 induce IL-17A secretion from human central memory CD4(+) T cells, TGF-beta and IL-21 uniquely promote the differentiation of human naive CD4(+) T cells into T(H)17 cells accompanied by expression of the transcription factor RORC2. These data will allow the investigation of this new population of T(H)17 cells in human inflammatory disease.

Figure 1. TGF-β and IL-21 promote T_H17 differentiation from naive CD4⁺ T cells

a, CD4⁺ T cells were sorted into populations enriched for naive or central memory T helper cells. b, IFN-γ and IL-17A secretion is shown from T cells stimulated for 7 days in the presence of the indicated cytokines. Standard deviation using T cells from three unrelated subjects is represented. TGF-β- and IL-21-induced IL-17 secretion is highly significant (P < 0.01). Tx, treatment. c, IL-17 secretion from naive T cells from six different donors is represented (mean±s.e.m.). d, Intracellular expression of IL-17 and IFN-γ from one of five experiments is shown.

Figure 2. TGF-β and IL-21 induce RORC2 in naive CD4⁺ T cells

a, mRNA expression levels (fold-induction relative to T cells without exogenous cytokines) of RORC2, Tbet, GATA3, IL23R and FOXP3 are shown after naive T cells were differentiated as indicated. b, Fold-induction±s.e.m. (n = 3) of IL21 and IL22 are represented. c, Mean expression±s.e.m. of IL17A and RORC2 are shown (n = 3) for naive T cells obtained from cord blood. d, Shown is intracytoplasmic staining of IL-17 and IFN-γ from cord blood naive T cells after 7 days of differentiation. Similar results were seen in another independent assay.