Immunogenicity and Efficacy of Live L. Tarentolae Expressing Kmp11-Ntgp96-Gfp Fusion As A Vaccine Candidate Against Experimental Visceral Leishmaniasis Caused By L. Infantum

Background: the aim of present study was to evaluate the protective efficacy of live recombinant l. tarentolae expressing kmp11-ntgp96-gfp fusion as candidates for live engineered recombinant vaccine against visceral leishmaniasis in balb/c mice. methods: kmp-11 and nt-gp96 genes cloned into the pjet1.2/blunt cloning vector and then into pegfp-n1 expression vector. the kmp-11, nt-gp96 and gfp fused in pegfp-n1 and subcloned into leishmanian plexsy-neo vector. finally, this construct was transferred to l. tarentolae by electroporation. tranfection was confirmed by sds-page, western blot, flow cytometry and rt-pcr. protective efficacy of this construct was evaluated as a vaccine candidate against visceral leishmaniasis. parasite burden, humoral and cellular immune responses were assessed before and at 4 weeks after challenge. results: kmp- nt-gp96-gfp fusion was cloned successfully into plexsy-neo vector and this construct successfully transferred to l. tarentolae. finding indicated that immunization with l. tarentolae tarentolae-kmp11-ntgp96-gfp provides significant protection against visceral leishmaniasis and was able to induce an increased expression of ifn-γ and igg2a. following challenge, a reduced parasite load in the spleen of the kmp11-ntgp96-gfp immunized group was detected. conclusion: the present study is the first to use a combination of a leishmania antigen with an immunologic antigen in live recombinant l. tarentolae and results suggest that l. tarentolae-kmp11-ntgp96-gfp could be considered as a potential tool in vaccination against visceral leishmaniasis and this vaccination strategy could provide a potent rout for future vaccine development.