Academic journal article Iranian Journal of Public Health

Protective Efficacy of Live Recombinant Leishmania Tarentolae Expressing Kmp-11 - Ntgp96-Gfp Fusion Combined with Naloxone as a Vaccine Candidate against Visceral Leishmaniasis

Academic journal article Iranian Journal of Public Health

Protective Efficacy of Live Recombinant Leishmania Tarentolae Expressing Kmp-11 - Ntgp96-Gfp Fusion Combined with Naloxone as a Vaccine Candidate against Visceral Leishmaniasis

Article excerpt

Background: The use of the non-pathogenic Leishmania tarentolae as a live vaccine vector to deliver specific Leishmania antigens is a recent approach.

Methods: L.infantum KMP-11 and NT-Gp96 cloned into the pJET1.2/blunt cloning vector and then into pEGFP-N1 expression vector and then the KMP-11, NT-Gp96 and GFP fused in pEGFP-N1 and subcloned into leishmanian pLEXSY-neo vector. Finally this construct was transferred to Leishmania tarentolae by electroporation. Tranfection was con- firmed by SDS-PAGE, WESTERN blot, flowcytometry and RT-PCR. Protective Efficacy of recombinant Leishmania taren- tolae Combined with Naloxone were tested in susceptible BALB/c mice. Both humoral and cellular immune responses were assessed before challenge and at 4 weeks after Leishma- nia infection.

Results: KMP- NT-Gp96-GFP Fusion was cloned success- fully into pLEXSY-neo vector and this construct was suc- cessfully transferred into Leishmania tarentolae. …

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