Kalantari N, Ghaffari S
Cellular and Molecular Biology Research Center, Babol University of Medical Sciences; Laboratory Sciences Group, Faculty of Paramedical, Babol University of Medical Sciences, Babol, Iran.
Iran J Parasitol. 2013 Apr;8(2):197-206.
Molecules expressed on the surface of infected erythrocytes (IE) with Plasmodium falciparum play important roles in malaria pathogenesis and immune evasion. Some of these molecules are specific adhesive ligands mediating adhesion of IE to the vascular endothelium. In the current study, the antigens exposed on the surface of IE with different isolates and various binding subpopulations of P. falciparum were studied.
A pooled hyper immune serum (HIS) from Malawian adults and eluted antibodies from the surface of the homologous and heterologous parasites were used. The parasite surface molecules were analyzed by Immuno-Gold-Silver enhancement (IGSE) and Western blotting. Mini-column cytoadherence method was used to select various parasite-binding subpopulations.
Surface antigens of all the isolates were recognized by HIS and high recognition of antigens was observed in all isolates with homologous eluted antibodies. Western blot analysis showed that the eluted antibodies reacted with a small subset of antigens compared with HIS. Three bands, PfEMP-1, were detected in the Triton X- insoluble fraction of the ICAM-1 binding subpopulation. Another interesting band was ∼ 52-55 kDa in various isolates of P. falciparum. This molecule as defined by its low molecular weight, Triton X-100 solubility, surface location and sensitivity to 1 mg/ml trypsin.
The IE's surface antigens differed in parental population compared with the selected subpopulations. These molecules could induce isolate-specific immunity. Antibodies purified from the surface of IE can be used as specific reagents to investigate parasite-derived proteins expressed on the surface of IE.
感染恶性疟原虫的红细胞(IE)表面表达的分子在疟疾发病机制和免疫逃避中起重要作用。其中一些分子是介导IE与血管内皮细胞黏附的特异性黏附配体。在本研究中,对不同分离株和恶性疟原虫不同结合亚群的IE表面暴露的抗原进行了研究。
使用来自马拉维成年人的混合超免疫血清(HIS)以及从同源和异源寄生虫表面洗脱的抗体。通过免疫金银增强法(IGSE)和蛋白质印迹法分析寄生虫表面分子。采用微柱细胞黏附法选择各种寄生虫结合亚群。
所有分离株的表面抗原均被HIS识别,并且在所有分离株中使用同源洗脱抗体时均观察到对抗原的高识别率。蛋白质印迹分析表明,与HIS相比,洗脱的抗体与一小部分抗原发生反应。在ICAM-1结合亚群的Triton X-不溶性部分中检测到三条带,即PfEMP-1。在恶性疟原虫的各种分离株中,另一条有趣的带约为52-55 kDa。该分子由其低分子量、Triton X-100溶解性、表面定位和对1 mg/ml胰蛋白酶的敏感性所定义。
与选定的亚群相比,IE的表面抗原在亲本群体中有所不同。这些分子可诱导分离株特异性免疫。从IE表面纯化的抗体可作为特异性试剂用于研究IE表面表达的寄生虫衍生蛋白。