嗜酸性粒细胞的形态变化与分泌。

Eosinophil Shape Change and Secretion.

机构信息

Division of Allergy, Asthma and Clinical Immunology, Mayo Clinic Arizona, Scottsdale, AZ, USA.

Alberta Respiratory Centre, Department of Medicine, University of Alberta, Edmonton, AB, Canada.

出版信息

Methods Mol Biol. 2021;2241:199-219. doi: 10.1007/978-1-0716-1095-4_17.

DOI:10.1007/978-1-0716-1095-4_17

PMID:33486739

Abstract

The analysis of eosinophil shape change and mediator secretion is a useful tool in understanding how eosinophils respond to immunological stimuli and chemotactic factors. Eosinophils undergo dramatic shape changes, along with secretion of the granule-derived enzyme eosinophil peroxidase (EPX) in response to chemotactic stimuli including platelet-activating factor (PAF) and CCL11 (eotaxin-1). Here, we describe the analysis of eosinophil shape change by confocal microscopy analysis and provide an experimental approach for comparing unstimulated cells with those that have been stimulated to undergo chemotaxis. In addition, we illustrate two different degranulation assays for EPX using OPD and an ELISA technique and show how eosinophil degranulation may be assessed from in vitro as well as ex vivo stimulation.

摘要

分析嗜酸性粒细胞的形态变化和介质分泌是了解嗜酸性粒细胞如何对免疫刺激和趋化因子做出反应的有用工具。嗜酸性粒细胞在血小板激活因子 (PAF) 和 CCL11（嗜酸性粒细胞趋化因子-1）等趋化刺激物的作用下会发生剧烈的形态变化，并分泌颗粒衍生的酶——嗜酸性粒细胞过氧化物酶 (EPX)。在这里，我们通过共聚焦显微镜分析来描述嗜酸性粒细胞形态变化的分析，并提供一种实验方法来比较未受刺激的细胞与那些已经被刺激进行趋化的细胞。此外，我们还使用 OPD 和 ELISA 技术说明了两种不同的 EPX 脱颗粒测定方法，并展示了如何从体外和体内刺激评估嗜酸性粒细胞脱颗粒。