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
|
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: | Schools > Medicine Research Institutes & Centres > 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 |
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