Measuring Naturally Acquired Phagocytosis-Inducing Antibodies to <em>Plasmodium falciparum</em> Parasites by a Flow Cytometry-Based Assay

Maria del Pilar Quintana; Nsoh  Godwin Anabire; Lars Hviid

doi:10.3791/61538

通过流细胞学测定，测量自然获得的噬菌体诱导恶性疟Plasmodium falciparum原虫抗体

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Measuring Naturally Acquired Phagocytosis-Inducing Antibodies to Plasmodium falciparum Parasites by a Flow Cytometry-Based Assay

通过流细胞学测定，测量自然获得的噬菌体诱导恶性疟Plasmodium falciparum原虫抗体

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09:57 min

August 6, 2020

DOI: 10.3791/61538-v

Maria del Pilar Quintana¹, Nsoh Godwin Anabire^1,2,3, Lars Hviid^1,4

¹Centre for Medical Parasitology, Department of Immunology and Microbiology, Faculty of Health and Medical Sciences,University of Copenhagen, ²West African Centre for Cell Biology of Infectious Pathogens, Department of Biochemistry, Cell and Molecular Biology,University of Ghana, ³Department of Immunology,Noguchi Memorial Institute for Medical Research, ⁴Centre for Medical Parasitology, Department of Infectious Diseases,Rigshospitalet

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该协议的总体目标是指导如何测量自然接触恶性疟原虫感染的血清或血浆中的抗体的能力，以对抗和诱导寄生虫感染的红细胞（IEs）的噬菌体。 Plasmodium falciparum

该协议允许测量抗体对恶性疟原虫感染红细胞的吞咽和诱导细胞病的能力。可测试自然接触寄生虫感染的个体血清中的抗体，以及由寄生虫抗原免疫引起的抗体。与我一起演示这个程序的将是我实验室的技术人员迈肯·维斯蒂。

为了建立一个THP-1细胞培养物进行噬菌体测定，在实验前一天将细胞播种在25厘米方培养瓶中，浓度为2.5倍，10至5.5细胞，在THP一个细胞培养基中。在实验当天，在实验开始前至少一小时，每孔PBS中用150微升无菌2%FBS的圆底96孔板进行圆底。对于中晚期受三磷酸盐感染的红细胞的收获和纯化，在10毫升寄生虫培养基中准备约1.2毫升的包装P恶性疟原虫培养物，至少含有5%的寄生虫，并加入磁柱。

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