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评估T细胞受体介导的人T细胞细胞毒性的方案。

Protocol for assessing T cell receptor-mediated human T cell cytotoxicity.

作者信息

Marks Kathryne E, Adejoorin Ifeoluwakiisi, Sowerby John, Rao Deepak A

机构信息

Brigham and Women's Hospital, Division of Rheumatology, Inflammation, and Immunity, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.

Brigham and Women's Hospital, Division of Rheumatology, Inflammation, and Immunity, Boston, MA, USA.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103541. doi: 10.1016/j.xpro.2024.103541. Epub 2025 Jan 4.

Abstract

Measuring the cytotoxicity of human T cells is a valuable but challenging task. We present a protocol for assessing the ability of polyclonal human T cells to kill target cells upon T cell receptor (TCR) triggering. We describe steps for preparing L and T cells for the assay and incubation of target cells with effector cells. We detail procedures for assessing cell death by flow cytometry. This protocol can be applied to small numbers of effector cells isolated from patient samples. For complete details on the use and execution of this protocol, please refer to Wang et al..

摘要

测量人类T细胞的细胞毒性是一项有价值但具有挑战性的任务。我们提出了一种方案,用于评估多克隆人类T细胞在T细胞受体(TCR)触发后杀死靶细胞的能力。我们描述了为该测定准备L细胞和T细胞以及靶细胞与效应细胞共孵育的步骤。我们详细说明了通过流式细胞术评估细胞死亡的程序。该方案可应用于从患者样本中分离出的少量效应细胞。有关该方案的使用和执行的完整详细信息,请参考Wang等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2918/11758416/d39b7d4f46e1/fx1.jpg

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