利用混合骨髓嵌合体测量真菌感染过程中的体内中性粒细胞迁移反应。

Measuring In Vivo Neutrophil Trafficking Responses During Fungal Infection Using Mixed Bone Marrow Chimeras.

机构信息

Fungal Pathogenesis Section, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.

Institute of Immunology and Immunotherapy, Institute of Microbiology and Infection, University of Birmingham, Birmingham, UK.

出版信息

Methods Mol Biol. 2021;2260:179-196. doi: 10.1007/978-1-0716-1182-1_12.

DOI:10.1007/978-1-0716-1182-1_12

PMID:33405038

Abstract

Neutrophil migration to the site of infection is an essential process for the control and clearance of microbial growth within the host. Identifying the molecular factors that mediate neutrophil chemotaxis is therefore critical for our understanding of disease pathogenesis and the mechanisms underlying protective immunity. Here, we describe a protocol that enables analysis of neutrophil recruitment from the blood into fungal-infected organs in vivo, using mixed bone marrow chimeras and flow cytometry. This method directly assesses the relative contribution of a receptor or intracellular molecule in controlling neutrophil chemotaxis during fungal infection and can be adapted to a variety of other non-fungal infection experimental settings.

摘要

中性粒细胞向感染部位的迁移是宿主控制和清除微生物生长的一个重要过程。因此，确定介导中性粒细胞趋化的分子因素对于我们理解疾病发病机制和保护性免疫的机制至关重要。在这里，我们描述了一种使用混合骨髓嵌合体和流式细胞术分析体内从血液向真菌感染器官募集中性粒细胞的方案。该方法直接评估了受体或细胞内分子在控制真菌感染期间中性粒细胞趋化作用中的相对贡献，并且可以适应各种其他非真菌感染实验条件。