Centrin-Deleted Leishmania donovani Parasites Help CD4(+) T Cells to Acquire Th1 Phenotype and Multi-Functionality Through Downregulation of CD200–CD200R Immune Inhibitory Axis

The protozoan parasite Leishmania has evolved several strategies to undermine host defense mechanisms by inducing Th2-type adaptive immunity and suppressing effector functions of Th1 phenotype. In our earlier studies, using centrin gene-deleted Leishmania (LdCen(−/−)) parasites as an immunogen, we have shown induction of an effective Th1-type immunity and robust memory responses that mediate protection against virulent challenge. However, role of inhibitory signals in Leishmania vaccine induced immunity in general, and LdCen(−/−) in particular has not been studied. Herein, we report that immunization with LdCen(−/−) parasites produces more functional Th1-type CD4(+) T cells via downregulation of CD200–CD200R immune inhibitory axis compared to wild-type infection. We found that expression of CD200 and CD200R was significantly reduced in LdCen(−/−) infection compared to wild-type infection. Diminished CD200–CD200R signaling in LdCen(−/−) infection enabled proliferation of CD4(+) T cells and resulted in the induction of pro-inflammatory cytokines and suppression of anti-inflammatory response. The effects of diminished CD200–CD200R signaling by LdCen(−/−) were most evident in the suppression of IL-10-producing CD4(+) T cells that helped enhance more Th1 cytokine producing and multi-functional T cells compared to wild-type infection. In vivo blocking of CD200 expression with anti-CD200 treatment in wild-type infected mice limited Th2 response as indicated by reduction of IL-10-producing Tr1 cells and reduced parasite burden. On the other hand, treatment with anti-CD200 improved the LdCen(−/−) vaccine-induced multifunctional response and reduction in splenic parasite load upon challenge. Taken together, these studies demonstrate the role of CD200–CD200R signals in the protection induced by LdCen(−/−) parasites.