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IFN- and IL-10 Induce the Differentiation of Human Type 1 T Regulatory Cells¹

Megan K. Levings^*, Romina Sangregorio^*, Francesca Galbiati^*, Stefania Squadrone^*, Rene de Waal Malefyt and Maria-Grazia Roncarolo^2,*

^* San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy; and DNAX Research Institute, Palo Alto, CA 94304

CD4⁺ T regulatory type 1 (Tr1) cells suppress Ag-specific immune responses in vitro and in vivo. Although IL-10 is critical for the differentiation of Tr1 cells, the effects of other cytokines on differentiation of naive T cells into Tr1 cells have not been investigated. Here we demonstrate that endogenous or exogenous IL-10 in combination with IFN-, but not TGF-, induces naive CD4⁺ T cells derived from cord blood to differentiate into Tr1 cells: IL-10⁺IFN-⁺IL-2^-/lowIL-4^-. Naive CD4⁺ T cells derived from peripheral blood require both exogenous IL-10 and IFN- for Tr1 cell differentiation. The proliferative responses of the Tr1-containing lymphocyte populations, following activation with anti-CD3 and anti-CD28 mAbs, were reduced. Similarly, cultures containing Tr1 cells displayed reduced responses to alloantigens via a mechanism that was partially mediated by IL-10 and TGF-. More importantly, Tr1-containing populations strongly suppressed responses of naive T cells to alloantigens. Collectively, these results show that IFN- strongly enhances IL-10-induced differentiation of functional Tr1 cells, which represents a first major step in establishing specific culture conditions to generate T regulatory cells for biological and biochemical analysis, and for cellular therapy to induce peripheral tolerance in humans.

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