In vitro generated Th17 cells support the expansion and phenotypic stability of CD4+Foxp3+ regulatory T cells in vivo

• Th17 cells express the highest levels of TNF than other Th subsets.
• Th17 cells most potently stimulate co-transferred Tregs in Rag1 KO mice.
• This effect of Th17 cells is TNF- and IL-2-dependent.
• TNF–TNFR2 plays crucial role in reciprocal stimulation of Th17 cells and Tregs.

CD4+ T cells stimulate immune responses through distinct patterns of cytokine produced by Th1, Th2 or Th17 cells, or inhibit immune responses through Foxp3-expressing regulatory T cells (Tregs). Paradoxically, effector T cells were recently shown to activate Tregs, however, it remains unclear which Th subset is responsible for this effect. In this study, we found that Th17 cells expressed the highest levels of TNF among in vitro generated Th subsets, and most potently promoted expansion and stabilized Foxp3 expression by Tregs when co-transferred into Rag1-/- mice. Both TNF and IL-2 produced by Th17 cells contributed to this effect. The stimulatory effect of Th17 cells on Tregs was largely abolished when co-transferred with TNFR2-deficient Tregs. Furthermore, Tregs deficient in TNFR2 also supported a much lower production of IL-17A and TNF expression by co-transferred Th17 cells. Thus, our data indicate that the TNF–TNFR2 pathway plays a crucial role in the reciprocal stimulatory effect of Th17 cells and Tregs. This bidirectional interaction should be taken into account when designing therapy targeting Th17 cells, Tregs, TNF and TNFR2.