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

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

doi:10.1016/j.xpro.2024.103541

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

STAR Protoc. 2025 Jan 4;6(1):103541. doi: 10.1016/j.xpro.2024.103541. Online ahead of print.

Authors

Kathryne E Marks¹, Ifeoluwakiisi Adejoorin², John Sowerby³, Deepak A Rao⁴

Affiliations

¹ Brigham and Women's Hospital, Division of Rheumatology, Inflammation, and Immunity, Boston, MA, USA; Harvard Medical School, Boston, MA, USA. Electronic address: [email protected].
² Brigham and Women's Hospital, Division of Rheumatology, Inflammation, and Immunity, Boston, MA, USA.
³ Brigham and Women's Hospital, Division of Rheumatology, Inflammation, and Immunity, Boston, MA, USA; Harvard Medical School, Boston, MA, USA.
⁴ Brigham and Women's Hospital, Division of Rheumatology, Inflammation, and Immunity, Boston, MA, USA; Harvard Medical School, Boston, MA, USA. Electronic address: [email protected].

PMID: 39756033
DOI: 10.1016/j.xpro.2024.103541

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.¹.

Keywords: Cell culture; Cell-based Assays; Immunology.