Generation of human Th1-like regulatory CD4+ T cells by an intrinsic IFN-γ- and T-bet-dependent pathway

Murine Foxp3(+) Treg have recently been shown to express T-bet, a transcription factor characteristic of Th1 effector cells. A human Treg phenotype equivalent has not been reported. Here, we show that naive human CD4(+) T cells incubated with low numbers of CD40-activated allogeneic B cells preferentially differentiate into alloantigen-specific CD4(hi)CD25(hi) Treg. These differentiated cells potently suppress effector T-cell responses and express T-bet, IFN-gamma, and CXCR3, the features of Th1 effector cells. In contrast, co-culture of naive CD4(+) T cells with high numbers of allogeneic B cells results in CD4(+)CD25(+) T cells that promote, rather than inhibit, effector T-cell responses, demonstrating the plasticity of CD4(+) T-cell differentiation in response to alloantigen-presenting B cells. The optimal accumulation of CD4(hi)CD25(hi) Treg induced using higher T cell: B cell co-culture ratios was dependent on the expression of T-bet and endogenously produced IFN-gamma. Induction of Treg-mediated suppression function in the Treg population was not. As CXCR3 confers the preferential trafficking of T cells to tissue sites of IFN-gamma, these human Th1-like Treg might be useful for modulating pathological Th1 effector responses, such as that occurring during graft-versus-host disease or graft rejection.