Techniques to improve the direct ex vivo detection of low frequency antigen-specific CD8+T cells with peptide-major histocompatibility complex class I tetramers

Chattopadhyay, Pratip K., Melenhorst, J. Joseph, Ladell, Kristin Ingrid ORCID: https://orcid.org/0000-0002-9856-2938, Gostick, Emma, Scheinberg, Phillip, Barrett, A. John, Wooldridge, Linda, Roederer, Mario, Sewell, Andrew K. ORCID: https://orcid.org/0000-0003-3194-3135 and Price, David A. ORCID: https://orcid.org/0000-0001-9416-2737 2008. Techniques to improve the direct ex vivo detection of low frequency antigen-specific CD8+T cells with peptide-major histocompatibility complex class I tetramers. Cytometry Part A 73A (11) , pp. 1001-1009. 10.1002/cyto.a.20642

Full text not available from this repository.

Abstract

The ability to quantify and characterize antigen-specific CD8+ T cells irrespective of functional readouts using fluorochrome-conjugated peptide-major histocompatibility complex class I (pMHCI) tetramers in conjunction with flow cytometry has transformed our understanding of cellular immune responses over the past decade. In the case of prevalent CD8+ T cell populations that engage cognate pMHCI tetramers with high avidities, direct ex vivo identification and subsequent data interpretation is relatively straightforward. However, the accurate identification of low frequency antigen-specific CD8+ T cell populations can be complicated, especially in situations where T cell receptor-mediated tetramer binding occurs at low avidities. Here, we highlight a few simple techniques that can be employed to improve the visual resolution, and hence the accurate quantification, of tetramer binding CD8+ T cell populations by flow cytometry. These methodological modifications enhance signal intensity, especially in the case of specific CD8+ T cell populations that bind cognate antigen with low avidities, minimize background noise, and enable improved discrimination of true pMHCI tetramer binding events from nonspecific uptake.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine Systems Immunity Research Institute (SIURI)
Subjects:	R Medicine > R Medicine (General)
Uncontrolled Keywords:	CD8+ T cell ; Peptide-major histocompatibility complex class I tetramer ; Polychromatic flow cytometry
Additional Information:	Special Issue: MASIR: Measurement of Antigen-Specific Immune Responses, 2008
Publisher:	Wiley-Blackwell
ISSN:	1552-4922
Last Modified:	05 Nov 2022 15:04
URI:	https://orca.cardiff.ac.uk/id/eprint/22303

Citation Data

Cited 46 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item