Assignment of functional roles to parasite proteins in malaria-infected red blood cells by competitive flow-based adhesion assay
Assignment of functional roles to parasite proteins in malaria-infected red blood cells by competitive flow-based adhesion assay
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DOI：
10.1046/j.1365-2141.2002.03404.x
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发表时间：
2002-04-01
影响因子：
6.5
通讯作者：
Coppel, RL
中科院分区：
文献类型：
--
作者：
Cooke, BM;Glenister, FK;Coppel, RL
被寄生的红细胞（PRBCs）粘附到内皮细胞并随后在微血管中积聚是恶性疟疾发病机制中的关键事件。在红细胞内发育过程中，寄生虫输出的许多蛋白质与RBC膜骨架相关，但这些蛋白质中的许多蛋白质的确切功能仍然未知。然而，它们的细胞位置表明，有些可能在粘附中发挥作用。PRBCs的粘附特性最好在体外流动条件下进行研究;然而，在目前可用的测定中，细胞粘附水平的实验变化使得粘附的细微变化难以量化。在这里，我们描述了一种基于流动的测定，可以量化粘附的微小差异，并记录了一些寄生虫蛋白质影响粘附的程度，使用不再表达特定蛋白质的寄生虫系。寄生虫蛋白环感染的红细胞表面抗原（RESA），旋钮相关的组氨酸丰富的蛋白（KAHRP）或恶性疟原虫红细胞膜蛋白3（PfEMP 3）的损失有一个显着的影响PRBCs的粘附能力，而成熟的寄生虫感染的红细胞表面抗原（梅萨）的损失没有影响。我们的研究表明，一些膜粘附相关的寄生虫蛋白，虽然没有暴露在红细胞表面，可以共同影响PRBCs的粘附特性，并进一步了解疟疾感染的红细胞的病理生理相关的结构/功能关系。
Adhesion of parasitized red blood cells (PRBCs) to endothelial cells and subsequent accumulation in the microvasculature are pivotal events in the pathogenesis of falciparum malaria. During intraerythrocytic development, numerous proteins exported from the parasite associate with the RBC membrane skeleton but the precise function of many of these proteins remain unknown. Their cellular location, however, suggests that some may play a role in adhesion. The adhesive properties of PRBCs are best studied under flow conditions in vitro ; however, experimental variation in levels of cytoadherence in currently available assays make subtle alterations in adhesion difficult to quantify. Here, we describe a flow-based assay that can quantify small differences in adhesion and document the extent to which a number of parasite proteins influence adhesion using parasite lines that no longer express specific proteins. Loss of parasite proteins ring-infected erythrocyte surface antigen (RESA), knob-associated histidine-rich protein (KAHRP) or Plasmodium falciparum erythrocyte membrane protein 3 (PfEMP3) had a significant effect on the ability of PRBCs to adhere, whereas loss of mature parasite-infected erythrocyte surface antigen (MESA) had no effect. Our studies indicate that a number of membrane skeleton-associated parasite proteins, although not exposed on the RBC surface, can collectively affect the adhesive properties of PRBCs and further our understanding of pathophysiologically relevant structure/function relationships in malaria-infected RBCs.