通过免疫磁珠去除 EpCAM 细胞来高效富集小鼠小肠上皮内淋巴细胞。

High-yield enrichment of mouse small intestine intraepithelial lymphocytes by immunomagnetic depletion of EpCAM cells.

机构信息

Experimental Immunology Branch, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA.

出版信息

STAR Protoc. 2022 Feb 25;3(1):101207. doi: 10.1016/j.xpro.2022.101207. eCollection 2022 Mar 18.

DOI:10.1016/j.xpro.2022.101207

PMID:35243383

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8885746/

Abstract

Lymphocytes in the mouse small intestine (SI) epithelium are critical to establish effective barrier immunity. Here, we describe a magnetic cell separation protocol that employs anti-epithelial cellular adhesion molecule (EpCAM) antibodies to deplete epithelial cells and to enrich for intraepithelial lymphocytes (IELs) from SI tissues. The resulting IEL preparation was functionally and phenotypically comparable to IELs isolated using conventional density gradient centrifugation protocols. Moreover, the yield and purity of anti-EpCAM-depleted SI IELs were higher than those enriched by conventional isolation. For complete details on the use and execution of this protocol, please refer to Prakhar et al. (2021).

摘要

从小鼠小肠（SI）上皮细胞中分离淋巴细胞对于建立有效的屏障免疫至关重要。在这里，我们描述了一种使用抗上皮细胞黏附分子（EpCAM）抗体的磁性细胞分离方案，该方案可用于耗尽上皮细胞并从小肠组织中富集上皮内淋巴细胞（IEL）。所得的 IEL 制剂在功能和表型上与使用常规密度梯度离心方案分离的 IEL 相当。此外，与传统的分离方法相比，使用抗 EpCAM 耗尽的 SI IEL 的产量和纯度更高。有关该方案使用和执行的完整详细信息，请参阅 Prakhar 等人（2021 年）。