Horowitz Amir, Riley Eleanor M
Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, UK.
Methods Mol Biol. 2013;923:447-64. doi: 10.1007/978-1-62703-026-7_31.
This chapter describes a protocol to assess activation of human NK cells following in vitro stimulation with malaria-infected red blood cells. Activation is assessed by flow cytometry, staining for cell surface expression of CD69 and accumulation of intracellular IFN-γ. Procedures are described for in vitro propagation and purification of Plasmodium falciparum parasites, separation of peripheral blood mononuclear cells from heparinized blood by density centrifugation, in vitro culture of PBMC and for staining and analysis of PBMC by flow cytometry. Some examples of typical FACS plots are shown.
本章描述了一种用于评估经疟疾感染的红细胞体外刺激后人自然杀伤细胞(NK细胞)激活情况的方案。通过流式细胞术、检测细胞表面CD69的表达以及细胞内γ干扰素(IFN-γ)的积累来评估激活情况。文中描述了恶性疟原虫寄生虫的体外增殖和纯化、通过密度离心从肝素化血液中分离外周血单核细胞、外周血单核细胞的体外培养以及通过流式细胞术对外周血单核细胞进行染色和分析的步骤。还展示了一些典型的荧光激活细胞分选(FACS)图示例。