从人外周血单核细胞 (PBMCs) 中分离和鉴定常规和非常规 2 型先天淋巴细胞 (ILC2s)。

Isolation and Characterization of Conventional and Non-conventional Type 2 Innate Lymphoid Cells (ILC2s) from Human Peripheral Blood Mononuclear Cells (PBMCs).

机构信息

Division of Allergy and Immunology, Department of Medicine, National Jewish Health, Denver, CO, USA.

Department of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.

出版信息

Methods Mol Biol. 2022;2506:187-198. doi: 10.1007/978-1-0716-2364-0_13.

DOI:10.1007/978-1-0716-2364-0_13

PMID:35771472

Abstract

Innate lymphoid cells (ILCs) are a relatively new family of lymphoid cells that lack lineage cell surface markers but produce various effector cytokines. Based on phenotype and function, the group 2 ILCs (ILC2s) mirror the features of the adaptive CD4 Th2 cell subset. In humans, they are traditionally characterized as the LinIL7RαCRΤΗ2CD161 cell population that produces type 2 cytokines - IL-5 and IL-13. However, the commonly used surface markers for human ILC2s leave a majority of type 2 cytokine-producing ILC2s unaccounted for. Recently, we characterized a distinct type 2 cytokine-producing Lin population that lacks surface expression of canonical CRTH2 but expresses CD30 and TNFR2. Herein, we describe a detailed protocol for isolation, staining, and analysis of the conventional LinCRTH2IL7Ra and the non-conventional LinCD30TNFR2 ILC2 populations.

摘要

先天淋巴细胞 (ILC) 是一组相对较新的淋巴细胞，缺乏谱系细胞表面标志物，但能产生各种效应细胞因子。根据表型和功能，第 2 组 ILC (ILC2) 与适应性 CD4 Th2 细胞亚群的特征相似。在人类中，它们传统上被定义为 LinIL7RαCRΤΗ2CD161 细胞群，能够产生 2 型细胞因子 - IL-5 和 IL-13。然而，常用的人类 ILC2 表面标志物未能涵盖大多数产生 2 型细胞因子的 ILC2。最近，我们鉴定了一种独特的 2 型细胞因子产生的 Lin 细胞群，该细胞群缺乏经典的 CRTH2 表面表达，但表达 CD30 和 TNFR2。在此，我们描述了一种详细的方案，用于分离、染色和分析传统的 LinCRTH2IL7Ra 和非传统的 LinCD30TNFR2 ILC2 群体。