title: Immunogenicity and Efficacy of LiveL. tarentolaeExpressing KMP11-NTGP96-GFP Fusion as a Vaccine Candidate against Experimental VisceralLeishmaniasisCaused byL. infantum.
abstract: BACKGROUND
  The aim of present study was to evaluate the protective efficacy of live recombinantL. tarentolaeexpressing 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 intoLeishmanian pLEXSY-neo vector. Finally this construct was transferred toL. tarentolaeby electroporation. Tranfection was confirmed by SDS-PAGE, WESTERN blot, flowcytometry 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 toL. tarentolae. Finding indicated that immunization withL. 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.
  
  
  CONCLUSIONS
  The present study is the first to use a combination of aLeishmaniaantigen with an immunologic antigen in live recombinantL. tarentolaeand results suggest thatL. 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.
authors.affiliation: Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Vahid
authors.initials: V
authors.lastname: Nasiri
authors.affiliation: Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Abdolhossein
authors.initials: A
authors.lastname: Dalimi
authors.affiliation: Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Ghaffarifar
authors.affiliation: Department of Hepatitis & AIDS, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Azam
authors.initials: A
authors.lastname: Bolhassani
keywords: KMP-11; L. infantum; L. tarentolae; NT-GP96; Vaccine; Visceral leishmaniasis