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There is no safe and efficacious vaccine against human leishmaniasis available and live attenuated vaccines have been used as a prophylactic alternative against the disease. In order to obtain an attenuated  Leishmania  parasite for vaccine purposes, we generated  L. infantum KHARON1  ( KH1 ) null mutants (Δ Likh1 ). This gene was previously associated with growth defects in  L. mexicana . Δ Likh1  was obtained and confirmed by PCR, qPCR and  Southern blot . We also generate a  KH1  complemented line with the introduction of episomal copies of  KH1 . Although Δ Likh1  promastigote forms exhibited a growth pattern similar to the wild-type line, they differ in morphology without affecting parasite viability.  L. infantum  KH1-deficient amastigotes were unable to sustain experimental infection in macrophages, forming multinucleate cells which was confirmed by  in vivo  attenuation phenotype. The cell cycle analysis of Δ Likh1  amastigotes showed arrested cells at G 2 /M phase. Δ Likh1- immunized mice presented reduced parasite burden upon challenging with virulent  L. infantum , when compared to naïve mice. An effect associated with increased  Li  SLA-specific IgG serum levels and IL-17 production. Thus, Δ Likh1  parasites present an infective-attenuated phenotype due to a cytokinesis defect, whereas it induces immunity against visceral leishmaniasis in mouse model, being a candidate for antileishmanial vaccine purposes.

Growth arrested live-attenuated Leishmania infantum KHARON1 null mutants display cytokinesis defect and protective immunity in mice