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Evaluation of transgenic Leishmania infantum expressing mLLO‐BAX‐SMAC in the apoptosis of the infected macrophages in vitro and in vivo
Author(s) -
Aghaei Maryam,
Khanahmad Hossein,
Aghaei Shahrzad,
Hosseini Sayed Mohsen,
Farahmand Mahin,
Hejazi Seyed Hossein
Publication year - 2020
Publication title -
parasite immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.795
H-Index - 75
eISSN - 1365-3024
pISSN - 0141-9838
DOI - 10.1111/pim.12726
Subject(s) - biology , leishmania infantum , transgene , apoptosis , leishmania , western blot , parasite hosting , flow cytometry , blot , spleen , microbiology and biotechnology , leishmaniasis , visceral leishmaniasis , immunology , gene , genetics , world wide web , computer science
Background Leishmaniasis is an important infectious disease that develops because of escaping parasite from the host immune system or preventing host macrophages apoptosis. Recently, the development of transgenic methods and the manipulation of the parasite genome has provided many advantages. So, in this study, the effect of the transgenic Leishmania infantum expressing mLLO‐BAX‐SMAC proteins was examined in accelerating host cell apoptosis. Method The entire coding sequence of designed codon‐optimized m LLO ‐ Bax ‐ Smac was cloned in the pLexyNeo2 vector and integrated downstream of the 18srRNA locus of L infantum genome by homologous recombination. Next, the expression of mLLO‐BAX‐SMAC fusion protein was evaluated by the Western blotting technique and the pathogenesis of transgenic parasite was surveyed in vitro and in vivo. Results The results of PCR and Western blot confirmed proper integration and expression of m LLO‐Bax‐Smac sequence into the 18srRNA locus of L infantum . Flow cytometry showed accelerating apoptosis of transgenic Leishmania ‐infected macrophages compared to wild‐type parasite. Also, transgenic parasites were less virulent as a fewer parasitic burden was found in the spleen and liver of transgenic‐infected mice compared to the control. Conclusion The data suggested that the transgenic L infantum expressing BAX‐SMAC can be used as an experimental model for developing vaccination against leishmaniasis.