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Currently no effective vaccine is available for human visceral leishmaniasis(VL) caused by  Leishmania donovani . Previously, we showed that centrin1 and p27gene deleted live attenuated  Leishmania  parasites ( Ld Cen1 −/−  and  Ld p27 −/− ) are safe, immunogenic and protective in animal models. Here, to assess the correlates of protection, we evaluated immune responses induced by  Ld Cen1 −/−  and  Ld p27 −/−  in human blood samples obtained from healthy, healed VL (HVL), post kala-azar dermal leishmaniasis(PKDL) and VL subjects. Both parasites infected human macrophages, as effectively as the wild type parasites. Further,  Ld Cen1 −/−  and  Ld p27 −/−  strongly stimulated production of pro-inflammatory cytokines including, IL-12, IFN-γ, TNF-α, IL-2, IL-6 and IL-17 in the PBMCs obtained from individuals with a prior exposure to  Leishmania  (HVL and PKDL). There was no significant stimulation of anti-inflammatory cytokines (IL-4 and IL-10). Induction of Th1 biased immune responses was supported by a remarkable increase in IFN-γ secreting CD4 +  and CD8 +  T cells and IL-17 secreting CD4 +  cells in PBMCs from HVL cases with no increase in IL-10 secreting T cells. Hence,  Ld Cen1 −/−  and  Ld p27 −/−  are promising as live vaccine candidates against VL since they elicit strong protective immune response in human PBMCs from HVL, similar to the wild type parasite infection, mimicking a naturally acquired protection following cure.

Gene deleted live attenuated Leishmania vaccine candidates against visceral leishmaniasis elicit pro-inflammatory cytokines response in human PBMCs