Abstract
Leishmania (L) major and L. tropica are the etiological agents of cutaneous leishmaniosis. Leishmania species cause a board spectrum of phenotypes. A small number of genes are differentially expressed between them that have likely an important role in the disease phenotype. Procyclic and metacyclic are two morphological promastigote forms of Leishmania that express different genes. The glutathione peroxidase is an important antioxidant enzyme that essential in parasite protection against oxidative stress and parasite survival. This study aimed to compare glutathione peroxidase (TDPX) gene expression in procyclic and metacyclic and also interspecies in Iranian isolates of L. major and L. tropica. The samples were cultured in Novy-Nicolle-Mc Neal medium to obtain the promastigotes and identified using PCR-RFLP technique. They were then grown in RPMI1640 media for mass cultivation. The expression level of TDPX gene was compared by Real-time PCR. By comparison of expression level, up-regulation of TDPX gene was observed (5.37 and 2.29 folds) in L. major and L. tropica metacyclic compared to their procyclic, respectively. Moreover, there was no significant difference between procyclic forms of isolates, while 3.05 folds up-regulation in metacyclic was detected in L. major compared L. tropica. Our data provide a foundation for identifying infectivity and high survival related factors in the Leishmania spp. In addition, the results improve our understanding of the molecular basis of metacyclogenesis and development of new potential targets to control or treatment and also, to the identification of species-specific factors contributing to virulence and pathogenicity in the host cells.
Highlights
Leishmaniasis is a global parasitic disease caused by various species of Leishmania (L) that refer to the variety of diseases including self-healing cutaneous leishmaniasis (CL) to fatal visceral leishmaniasis (VL) [1]
The relative expression level of the TDPX gene between each specific stage of the L. major and L. tropica isolates was investigated, and no significant differences were detected between procyclic stages of two studied species (Fig. 3a), while TDPX gene expression level difference was significant between their metacyclic stages
Leishmania parasites pass through different developmental phases including procyclic promastigote, metacyclic promastigote and amastigote forms in mammalian host cells with an alteration in gene expression profile and morphological changes
Summary
Leishmaniasis is a global parasitic disease caused by various species of Leishmania (L) that refer to the variety of diseases including self-healing cutaneous leishmaniasis (CL) to fatal visceral leishmaniasis (VL) [1]. Procyclic and metacyclic are two morphological promastigote forms of Leishmania that express different genes. This study aimed to compare glutathione peroxidase (TDPX) gene expression in procyclic and metacyclic and interspecies in Iranian isolates of L. major and L. tropica. Methods: The samples were cultured in Novy-Nicolle-Mc Neal medium to obtain the promastigotes and identified using PCR-RFLP technique. They were grown in RPMI1640 media for mass cultivation. Results: By comparison of expression level, up-regulation of TDPX gene was observed (5.37 and 2.29 folds) in L. major and L. tropica metacyclic compared to their procyclic, respectively. There was no significant difference between procyclic forms of isolates, while 3.05 folds up-regulation in metacyclic was detected in L. major compared L. tropica. The results improve our understanding of the molecular basis of metacyclogenesis and development of new potential targets to control or treatment and to the identification of speciesspecific factors contributing to virulence and pathogenicity in the host cells
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