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Effect of Transgenic Leishmania Major Expressing Mllo-Bax-Smac Fusion Gene in the Apoptosis of the Infected Macrophages Publisher

Summary: Transgenic L. major with mLLO-BAX-SMAC accelerates macrophage apoptosis, reducing leishmaniasis progression. #Leishmaniasis #Vaccine

Aghaei M1 ; Khanahmad H2 ; Jalali A3 ; Aghaei S4 ; Narimani M5, 7 ; Hosseini SM6 ; Namdar F5, 7 ; Hejazi SH1, 8
Authors

Source: Iranian Journal of Basic Medical Sciences Published:2021


Abstract

Objective(s): Leishmaniasis is a complex infection against which no confirmed vaccine has been reported so far. Transgenic expression of proteins involved in macrophage apoptosis-like BAX through the parasite itself accelerates infected macrophage apoptosis and prevents Leishmania differentiation. So, in the present research, the impact of the transgenic Leishmania major including mLLO-BAX-SMAC proapoptotic proteins was assayed in macrophage apoptosis acceleration. Materials and Methods: The coding sequence mLLO-Bax-Smac was designed and integrated into the pLexyNeo2 plasmid. The designed sequence was inserted under the 18srRNA locus into the L. major genome using homologous recombination. Then, mLLO-BAX-SMAC expression was studied using the Western blot, and the transgenic parasite pathogenesis was investigated compared with wild-type L. major in vitro and also in vivo. Results: Western blot and PCR results approved mLLO-BAX-SMAC expression and proper integration of the mLLO-Bax-Smac fragment under the 18srRNA locus of L. major, respectively. The flow cytometry results revealed faster apoptosis of transgenic Leishmania-infected macrophages compared with wild-type parasite-infected macrophages. Also, the mild lesion with the less parasitic burden of the spleen was observed only in transgenic Leishmania-infected mice. The delayed progression of leishmaniasis was obtained in transgenic strain-injected mice after challenging with wild-type Leishmania. Conclusion: This study recommended transgenic L. major including mLLO-BAX-SMAC construct as a pilot model for providing a protective vaccine against leishmaniasis. © 2021 Mashhad University of Medical Sciences. All rights reserved.
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