Centre for Virus Research, The Westmead Institute for Medical Research, Westmead, Australia.
The Westmead Clinical School, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
Cytometry A. 2023 Nov;103(11):851-856. doi: 10.1002/cyto.a.24782. Epub 2023 Sep 29.
There is a great need to understand human immune cells within tissue, where disease manifests and infection occurs. Tissue-resident memory T cells (TRMs) were discovered over a decade ago, there is a great need to understand their role in human disease. We developed a 24-color flow cytometry panel to comprehensively interrogate CD4 and CD8 TRMs isolated from human tissues. When interrogating cells within human tissue, enzymatic methods used to liberate cells from within the tissue can cause cleavage of cell surface markers needed to phenotype these cells. Here we carefully select antibody clones and evaluate the effect of enzymatic digestion on the expression of markers relevant to the identification of T cell residency, as well as markers relevant to the activation and immunoregulation status of these cells. We have designed this panel to be applicable across a range of human tissues including skin, intestine, and type II mucosae such as the vagina.
非常有必要了解组织中出现疾病和感染的人类免疫细胞。组织驻留记忆 T 细胞 (TRM) 十多年前被发现，因此非常有必要了解它们在人类疾病中的作用。我们开发了一个 24 色流式细胞术面板，以全面研究从人体组织中分离出的 CD4 和 CD8 TRM。在研究人体组织中的细胞时，用于从组织中释放细胞的酶法会导致用于表型鉴定这些细胞的细胞表面标志物的裂解。在这里，我们仔细选择抗体克隆，并评估酶消化对与 T 细胞驻留鉴定相关的标记物以及与这些细胞的激活和免疫调节状态相关的标记物的表达的影响。我们设计了这个面板，使其可适用于包括皮肤、肠道和 II 型黏膜（如阴道）等多种人体组织。
