Live Attenuated <i>Leishmania donovani</i> Centrin Gene–Deleted Parasites Induce IL-23–Dependent IL-17–Protective Immune Response against Visceral Leishmaniasis in a Murine Model

Abstract

No vaccine exists against visceral leishmaniasis. To develop effective vaccines, we have previously reported protective role of live attenuated centrin gene-deleted <i>Leishmania donovani</i> (<i>LdCen^-/-</i> ) parasites through induction of Th1 type immune response in mice, hamsters, and dogs. In this study, we specifically explored the role of Th17 cells in <i>LdCen^-/-</i> -induced host protection in mice. Our results showed that compared with wild-type <i>L. donovani</i> infection, <i>LdCen^-/-</i> parasites induce significantly higher expression of Th17 differentiation cytokines in splenic dendritic cells. There was also induction of IL-17 and its promoting cytokines in total splenocytes and in both CD4 and CD8 T cells following immunization with <i>LdCen^-/-</i> Upon challenge with wild-type parasites, IL-17 and its differentiating cytokines were significantly higher in <i>LdCen^-/-</i> -immunized mice compared with nonimmunized mice that resulted in parasite control. Alongside IL-17 induction, we observed induction of IFN-γ-producing Th1 cells as reported earlier. However, Th17 cells are generated before Th1 cells. Neutralization of either IL-17 or IFN-γ abrogated <i>LdCen^-/-</i> -induced host protection further confirming the essential role of Th17 along with Th1 cytokines in host protection. Treatment with recombinant IL-23, which is required for stabilization and maintenance of IL-17, heightened Th17, and Tc17 responses in immunized mice splenocytes. In contrast, Th17 response was absent in immunized IL-23R^-/- mice that failed to induce protection upon virulent <i>Leishmania</i> challenge suggesting that IL-23 plays an essential role in IL-17-mediated protection by <i>LdCen^-/-</i> parasites. This study unveiled the role of IL-23-dependent IL-17 induction in <i>LdCen^-/-</i> parasite-induced immunity and subsequent protection against visceral leishmaniasis.