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The Journal of Immunology, 2005, 174: 1446-1455.
Copyright © 2005 by The American Association of Immunologists

Differentiation and Expansion of T Cells with Regulatory Function from Human Peripheral Lymphocytes by Stimulation in the Presence of TGF-{beta}

Patricia E. Rao1, Andria L. Petrone and Paul D. Ponath

TolerRx, Cambridge, MA 02139

T cells with immunoregulatory function have been described in human and mouse systems. In both systems these cells can be differentiated either in the thymus or from peripheral T cells. To date, more progress has been made in the study of murine regulatory T cells, because it has been very difficult to isolate human regulatory T cells of sufficient purity and in sufficient numbers to permit detailed examinations of their biochemistry. We report in this study that human T cells with regulatory function can be differentiated in vitro from naive (CD4+CD45RA+) cord blood or peripheral T cells by stimulation with anti-CD3 and anti-CD28 in the presence of TGF-{beta}. Cells derived in this manner express a surface phenotype (CD25+, CD122+, HLA-DR+, glucocorticoid-induced TNF receptor-related gene+, CD103+, CTLA-4+) described for human and mouse regulatory T cells and express protein and message for the transcription factor forkhead/winged helix transcription factor (FOXP3). They produce primarily TGF-{beta} and IL-10, with lesser amounts of IFN-{gamma} and IL-13, when stimulated through their TCRs and are capable of inhibiting cytokine production and proliferation by stimulated naive T cells. Unlike Th1 and Th2 cells, these TGF-{beta}-derived regulatory T cells do not appear to be dependent on the protein kinase C{theta} pathway of NF-{kappa}B activation for Ag-induced responses.




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