ELISPOT and functional T cell analyses using HLA mono-specific target cells

Horlock, Claire, Stott, Bryony, Dyson, Julian, Ogg, Graham, McPherson, Tess, Jones, Louise, Sewell, Andrew K. ORCID: https://orcid.org/0000-0003-3194-3135, Wooldridge, Linda, Cole, David ORCID: https://orcid.org/0000-0003-0028-9396, Stebbing, Justin and Savage, Philip 2009. ELISPOT and functional T cell analyses using HLA mono-specific target cells. Journal of Immunological Methods 350 (1-2) , pp. 150-160. 10.1016/j.jim.2009.08.011

Full text not available from this repository.

Abstract

Simple T cell assays specific for any chosen HLA class I or class II/peptide combination, are of enormous value in cancer immunotherapy, clinical trials, vaccine and infectious disease research. The reliable measurement of T cell activity can be difficult due to the presence of other alleles on target cells, particularly for the non-HLA-A2 alleles, and the varying baseline characteristics of the different APCs employed. In the absence of pulsing with HLA-A2 restricted peptides, T2 cells are functionally HLA class I and II negative. By coating these cells with recombinant HLA peptide complexes, HLA mono-specific cells are produced that present only a defined single epitope, and generate minimal background immune activation. In ELISPOT, intracellular cytokine staining (ICS) and killing assays using T cells specific for HLA-A2/peptide complexes, the HLA mono-specific cells gave comparable results, to those using standard peptide pulsed HLA-A2 positive T2 cells without significant background. Successful T cell assays for non-HLA-A2 T cells were also performed, with PBMCs recognizing HLA-A24 and HLA-DR15/peptide complexes. The data, obtained with ELISPOT, ICS and FACS-based killing assays, all demonstrate high specificity of T cell activity and low levels of background activity. HLA mono-specific cells are simple to prepare, and can be used with any stable recombinant HLA allele/peptide combination; providing a useful system for improved T cell functional analyses across all HLA allotypes. This represents a significant advance in the generation of reliable functional T cell data.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Systems Immunity Research Institute (SIURI)
Subjects:	Q Science > QR Microbiology > QR180 Immunology
Uncontrolled Keywords:	ELISPOT, cytotoxicity, T cell, assays, in vitro, immunotherapy
Publisher:	Elsevier
ISSN:	0022-1759
Last Modified:	20 Oct 2022 07:40
URI:	https://orca.cardiff.ac.uk/id/eprint/26016

Citation Data

Cited 2 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item