Department of Immunology and Microbiology, Faculty of Health and Medical Sciences, Centre for Medical Parasitology, University of Copenhagen, Copenhagen, Denmark.
Methods Mol Biol. 2022;2470:537-543. doi: 10.1007/978-1-0716-2189-9_41.
The var genes encoding the Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) mediating adhesion of infected erythrocytes (IEs) are clonally variant and confer the parasite the ability to evade the host immune response by enabling switching among expression of different PfEMP1 variants. This method of antigenic variation allows the parasite to adhere to a variety of proteins to escape splenic clearance. Enriching IE populations for expression of a specific PfEMP1 variant is crucial for the study of specific var genes PfEMP1 proteins and their role in pathogenicity. Selection of parasites using cell monolayers requires large-volume cultures (>5 mL). This chapter describes a method, which rapidly selects for adhesive phenotypes of interest using small-volume cultures (<1 mL) under physiological flow conditions.
编码介导感染红细胞(IEs)黏附的恶性疟原虫红细胞膜蛋白 1(PfEMP1)的 var 基因是克隆变异的,使寄生虫能够通过在不同 PfEMP1 变体的表达之间切换来逃避宿主免疫反应。这种抗原变异方法允许寄生虫附着在各种蛋白质上,以逃避脾脏清除。富集表达特定 PfEMP1 变体的 IE 群体对于研究特定 var 基因 PfEMP1 蛋白及其在致病性中的作用至关重要。使用细胞单层选择寄生虫需要大容量培养(>5 毫升)。本章描述了一种方法,该方法可在生理流动条件下使用小体积培养(<1 毫升)快速选择感兴趣的黏附表型。