Isolation of viable antigen-specific CD8+ T cells based on membrane-bound tumor necrosis factor (TNF)-α expression

Haney, Danielle, Quigley, Máire F., Asher, Tedi E., Ambrozak, David R., Gostick, Emma, Price, David ORCID: https://orcid.org/0000-0001-9416-2737, Douek, Daniel C. and Betts, Michael R. 2011. Isolation of viable antigen-specific CD8+ T cells based on membrane-bound tumor necrosis factor (TNF)-α expression. Journal of Immunological Methods 369 (1-2) , pp. 33-41. 10.1016/j.jim.2011.04.003

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Abstract

Current technology to isolate viable cytokine-producing antigen-specific primary human T cells is limited to bi-specific antibody capture systems, which suffer from limited sensitivity and high background. Here, we describe a novel procedure for isolating antigen-specific human T cells based on their ability to produce tumor necrosis factor (TNF)-α. Unlike many cytokines, TNF-α is initially produced in a biologically active membrane-bound form that is subsequently cleaved by TNF-α converting enzyme (TACE) to release the soluble form of TNF-α. By preventing this cleavage event, we show that TNF-α can be ‘trapped’ on the surface of the T cells from which it originates and directly labeled for viable isolation of these antigen-specific T cells. Together with other existing sorting procedures to isolate activated T cells, this new technique should permit the direct isolation of multi-functional T lymphocytes for further protein and gene expression analyses, as well as a detailed functional assessment of the potential role that TNF-α producing T cells play in the adaptive immune system.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Systems Immunity Research Institute (SIURI)
Subjects:	Q Science > QR Microbiology > QR180 Immunology R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer)
Uncontrolled Keywords:	cell sorting, T cells, TNF-α, viable, functional
Publisher:	Elsevier
ISSN:	0022-1759
Date of First Compliant Deposit:	19 January 2018
Last Modified:	15 Nov 2024 22:15
URI:	https://orca.cardiff.ac.uk/id/eprint/26017

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