Samant M, Sahasrabuddhe A A, Singh N, Gupta S K, Sundar S, Dube A
Divisions of Parasitology Central Drug Research Institute, Lucknow, India.
Parasitology. 2007 Aug;134(Pt 9):1175-84. doi: 10.1017/S0031182007002569. Epub 2007 Mar 16.
Leishmania produce several types of mucin-like glycoproteins called proteophosphoglycans (PPGs) some of which are secreted while others are found on the surface of promastigotes and amastigotes. These proteins are thought to be important in the transmission, invasion and subsequent intracellular survival of parasites. The structure and function of PPGs are species and stage-specific in the case of L. major and L. mexicana, but no such information has hitherto been available for L. donovani. This study presents, for the first time, an initial characterization (localization) of PPG in sodium stibogluconate (SSG)-resistant and sensitive clinical isolates of L. donovani from Bihar (India) by confocal microscopy, flow cytometry and Western blotting using antibodies to L. major PPG. Confocal microscopy analysis revealed that both promastigotes and amastigotes possess PPG on their cell membrane and flagellar pocket membrane but its expression was variable in different isolates. The quantitative analysis by FACS and Western blotting showed that the expression and intensity of PPG bands was higher in SSG-resistant isolates. This study suggests the possibilities of involvement of PPG in drug-resistant mechanisms and of using PPG abundance as a marker for identifying drug-resistant clinical isolates in Indian kala azar.
利什曼原虫产生几种类型的黏蛋白样糖蛋白,称为蛋白磷酸聚糖(PPGs),其中一些是分泌型的,而另一些则存在于前鞭毛体和无鞭毛体的表面。这些蛋白质被认为在寄生虫的传播、侵袭及随后的细胞内存活中起重要作用。对于硕大利什曼原虫和墨西哥利什曼原虫而言,PPGs的结构和功能具有种属和阶段特异性,但迄今尚无关于杜氏利什曼原虫的此类信息。本研究首次通过共聚焦显微镜、流式细胞术以及使用针对硕大利什曼原虫PPG的抗体进行蛋白质印迹法,对来自印度比哈尔邦的杜氏利什曼原虫抗葡萄糖酸锑钠(SSG)和敏感临床分离株中的PPG进行了初步表征(定位)。共聚焦显微镜分析显示,前鞭毛体和无鞭毛体在其细胞膜和鞭毛袋膜上均有PPG,但不同分离株中其表达存在差异。流式细胞术和蛋白质印迹法的定量分析表明,抗SSG分离株中PPG条带的表达和强度更高。本研究提示了PPG参与耐药机制的可能性,以及利用PPG丰度作为鉴定印度黑热病耐药临床分离株标志物的可能性。