The contribution of CD4+ CD25+ T-regulatory-cells to immune suppression in sepsis

Studies have indicated that there is a development of generalized immune dysfunction after septic insult. However, the mechanisms responsible for these changes remain unclear. Recently, accumulating evidence shows that several lymphocyte subpopulations such as NKT-, CD4(+)-Th2-T-, CD8(+)-T-, gamma delta T-, and CD4(+)CD25(+) T regulatory cells are capable of actively contributing to the induction of septic immune suppression. Thus, our aim was to investigate the contribution of CD4(+)CD25(+) cells to the immune dysfunction seen in sepsis. To study this, C57BL/6J, C57BL/6-116(tm1Kopf) (interleukin [IL] 6 -/-), and C57BL/6-ll10(tm1Cgn) (IL-10 -/-) mice were subjected to cecal ligation and puncture (CLP) or sham operations. Twenty-four hours later, blood was collected, and splenocytes were isolated. Phenotypic expression of CD4/CD25 (by fluorescence-activated cell sorter), cell proliferation (presented as proliferation index = [with anti-CD3]/ [without anti-CD3]), and immune suppressive capacity (by in vitro add-back experiments) were assessed. The results indicate a marked elevation in CD4(+)CD25(+) cell levels and their proliferation index after sepsis in background mice. CD4(+)CD25(-) cells from sham and CLP mice proliferated equally. However, coculture of CD4(+)CD25(-) with CD4(+)CD25(+) cells suppressed their proliferation in both sham and CLP mice. Depletion of CD25(+) cells in vivo before CLP markedly restored CD4(+)CD25(-) proliferative capacity and Th1 cytokine release while not altering plasma proinflammatory cytokine levels. Subsequently, IL-6 -/- and IL-10 -/- mice were used to elucidate the possible mediator(s) regulating the changes seen after sepsis. Although CD4(+)CD25(+) cells increased after septic insult in both C57BL/6J and IL-6 -/- mice, this was not observed in IL-10 -/- mice. Similarly, in vitro proliferation studies showed that proliferation index increased in CD4(+)CD25(+) cells from septic C57BL/6J and IL6 -/- mice, but it remained the same in IL-10 -/- mice. Surprisingly, depletion of CD25(+) cells before inducing sepsis did not alter septic mortality. Together, these findings suggest that although CD4(+)CD25(+) T regulatory cells induced by IL-10 seem to contribute to aspects of sepsis-induced lymphoid immune suppression, the oblation of CD25(+) cells does not provide a survival advantage or disadvantage.