Infection and Drug Resistance (Jul 2020)
Evaluation of the Anti-Leishmanial Effect of Recombinant Clostridium α-Toxin
Abstract
Fatemeh Namdar,1 Hossein Khanahmad,2 Zahra Ghayour,1 Farzaneh Mirzaei,3 Azam Namdar,4 Maryam Aghaei,5 Shahrokh Izadi,6 Faham Khamesipour,7,8 Seyed Hossein Hejazi1,9 1Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran; 2Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran; 3Department of Parasitology and Mycology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran; 4Department of Community Medicine, School of Medicine, Fasa University of Medical Sciences, Fasa, Iran; 5Skin Diseases and Leishmaniasis Research Center, Isfahan University of Medical Sciences, Isfahan, Iran; 6Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran; 7Cellular and Molecular Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran; 8Shahid Beheshti University of Medical Sciences, Tehran, Iran; 9Skin Diseases and Leishmaniasis Research Center, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranCorrespondence: Seyed Hossein HejaziSkin Diseases and Leishmaniasis Research Center, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, IranTel +98 9133118711Email [email protected]: Leishmaniasis is an infectious disease common in tropical and subtropical regions caused by the genus Leishmania, which is transmitted by the bite of female sandflies. In this study, we evaluate the anti-leishmanial effect of recombinant Clostridium α-toxin protein alone and the combination with glucantime through in vitro and in vivo.Materials and Methods: Production, expression, and purification of recombinant α-toxin were evaluated by SDS-PAGE and Western blotting techniques. The antileishmanial activities of the purified α-toxin plus and without glucantime were examined in vitro and in vivo.Results: The results indicated successful expression of α-toxin as a 48 kDa band on SDS-PAGE and Western blot methods. Also, evaluation of α-toxin IC50 showed the strong fatal effect of it, and glucantime on medium proliferated Leishmania promastigotes at lower concentrations compared with glucantime or α-toxin alone. Moreover, in vivo surveys showed that at the end of treatment courses, the mean of lesion size diminished in glucantime plus α-toxin treated mice versus negative control groups (p < 0.001). Also, there was a significant difference in the parasite burden of the spleen and liver of the control versus the test groups (p < 0.001).Conclusion: The results showed recombinant α-toxin has synergistic effects with glucantime in destroying Leishmania parasites.Keywords: α-toxin, Clostridium septicum, parasitic diseases, Leishmania, in vitro, in vivo
