Evaluation of Apoptotic Effect of Transgenic Leishmania tropica Expressing MLLO-Bax-Smac Fusion Gene in Infected Macrophages, an In Vitro Study

Abstract

Background:In the leishmaniasis parasitic infection, parasite escapesfrom the immune system of host or prevents macrophage apoptosis. It seems generating transgenic para-sites to express proapoptotic proteins can accelerate the apoptosis of infected macro-phage and prevent Leishmaniadifferentiation. Hence, weinvestigated the efficacy of trans-genic L. tropica expressing mLLO-BAX-SMACin expediting macrophage apoptosis.Methods:This study was performed at the Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences (2022 to 2023), Isfahan, Iran. mLLO-Bax-Smaccoding sequence cloned in the pLexyNeo2 was entered into the L. tropi-ca genomeusing homologous recombination. PCR, Western blot, and hemolysis tests were used to confirm integration accuracy. In addition, the apoptosis percentage of trans-genic L. tropica-infected macrophages was assayed by flow-cytometry.Results:The proper integration of the mLLO-Bax-Smacfragment into the 18srRNA lo-cus ofL. tropica and mLLO -BAX-SMAC fusion protein expression was approved. Fur-thermore, results of flow cytometry showed the mean percentage of apoptosis among thegroupsis different and transgenic L. tropicaleads to a decrease in the apoptosis time of infected macrophages compared to the wild type of Leishmaniaand to an increase in the immune system response. Conclusion:The data suggest the transgenic L. tropica can be used as an experimental model to expedite apoptosis of Leishmania-infected macrophage