Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
School of Pharmacy, Pacific University, Hillsboro, Oregon, USA.
BMC Vet Res. 2021 Apr 14;17(1):162. doi: 10.1186/s12917-021-02816-x.
Leishmania parasites express various essential proteins in different growth phases (logarithmic/stationary) and forms (promastigote/amastigote). Targeting the genes encoding such proteins paves the way for controlling these parasites. Centrin is an essential gene, which its protein product seems to be vital for the proliferation of Leishmania parasites. Herein, this study was contrived to analyze the expression level of the centrin gene in different growth phases and forms of Leishmania infantum (L. infantum) parasites isolated from various endemic areas of canine leishmaniasis (CanL) in Iran.
All three collected isolates were identified as L. infantum using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Real-time reverse transcription (RT)-PCR revealed a statistically significant up-regulation (3.13-fold) in the logarithmic phase promastigotes compared to stationary ones (p < 0.01), whereas centrin was expressed equally in intracellular amastigotes at different time points during cell culture. Also, our finding revealed a slight up-regulation of the centrin gene (1.22-fold) in the intracellular amastigotes compared to logarithmic phase promastigotes, which was found statistically non-significant (p > 0.05).
Centrin gene in Iranian isolates of L. infantum is more expressed in exponential than stationary phases and seems to be considered as a promising target in the development of a genetically modified live attenuated vaccine for CanL control.
利什曼原虫在不同的生长阶段(对数/静止)和形式(前鞭毛体/无鞭毛体)中表达各种必需蛋白。针对编码这些蛋白的基因为控制这些寄生虫铺平了道路。中心体是一个必需基因,其蛋白产物似乎对利什曼原虫的增殖至关重要。在此,本研究旨在分析从伊朗不同犬利什曼病(CanL)流行地区分离的利什曼原虫(L. infantum)不同生长阶段和形式的中心体基因的表达水平。
所有三个采集的分离株均通过聚合酶链反应-限制性片段长度多态性(PCR-RFLP)鉴定为 L. infantum。实时逆转录(RT)-PCR 显示对数期前鞭毛体与静止期相比表达上调(3.13 倍)(p < 0.01),而在细胞培养过程中不同时间点的内阿米巴体内中心体表达相等。此外,我们的研究结果表明,与对数期前鞭毛体相比,内阿米巴体中的中心体基因略有上调(1.22 倍),但统计学上无显著差异(p > 0.05)。
在伊朗 L. infantum 分离株中,中心体基因在指数生长期的表达高于静止期,因此被认为是开发用于控制 CanL 的基因修饰活减毒疫苗的有希望的靶标。