A role for trichosanthin in the expansion of CD4CD25 regulatory T cells

CD4(+)CD25(+) regulatory T cells (Tregs) are critical for the peripheral immune tolerance. Understanding the signals for the generation of Tregs is important for the clinical immunotherapy, but only limited progress has been made on obtaining enough peripheral Tregs. The aim of this study was to evaluate the role of trichosanthin (Tk) extracted from Chinese medicinal herb Trichosanthes kirilowi on the function of Tregs in vitro and in vivo. We reported here that Tk is needed for the expansion of freshly isolated CD4(+)CD25(+)Tregs (nTregs) into Tk-expanded CD4(+)CD25(+)Tregs (Tk-Tregs) through up-regulating CD25 and Foxp3 expression. The dose-response analyses indicated that 100 ng/ml Tk was the most appropriate dose. The result of real-time PCR showed that Tk-Tregs expressed 1.5-fold higher levels of Foxp3 than those observed in nTregs. Tk-Tregs markedly suppressed activation of effector T cells at a suppressor/responder ratio of 1:1, 1:2, 1:4, 1:8 or 1:16, and their effect was dose dependent. Moreover, Tk-Tregs secreted more immunosuppressive cytokines interleukin (IL)-10 and transforming growth factor (TGF)-beta1 after stimulating with antigen and antigen-presenting cells (APC). Transwell experiments showed that not only cell-to-cell contact but also soluble cytokines were involved in suppressive mechanism of Tk-Tregs. And Tk-Tregs were more efficient in suppressing CD25(-)T cell response to specific antigen than to irrelative antigen. Most importantly, it was revealed for the first time that Tk-Tregs could prolong the survival duration of mice with acute graft-versus-host disease (aGVHD). In conclusion, the study suggests a possible therapeutic potential of Tk-Tregs for clinical treatment on aGVHD.