Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 338, Issue 1-2, Pages 31-39Publisher
ELSEVIER
DOI: 10.1016/j.jim.2008.07.008
Keywords
peptide-major histocompatibility complex class I tetramer; polychromatic flow cytometry; T cell
Categories
Funding
- Intramural Research Program of the National Institutes of Health
- Vaccine Research Center
- National Institute of Allergy and Infectious Diseases
- National Heart, Lung and Blood Institute
- Medical Research Council (UK) Senior Clinical Fellow
- Wellcome Trust (UK) Clinical Intermediate Fellow
- Medical Research Council [G0501963] Funding Source: researchfish
- MRC [G0501963] Funding Source: UKRI
Ask authors/readers for more resources
The development of soluble recombinant peptide-major histocompatibility complex class I (pMHCI) molecules conjugated in multimeric form to fluorescent labels has enabled the physical quantification and characterization of antigen-specific CD8(+) T cell populations by flow cytometry. Several factors determine the binding threshold that enables visualization of cognate CD8(+) T cells with these reagents; these include the affinity of the T cell receptor (TCR) for pMHCl antigen. Here, we show that multimers constructed from peptide-human leukocyte antigen (pHLA) A*0201 monomers engineered in the heavy chain alpha 2 domain to enhance CD8 binding (K-D approximate to 85 mu M) without impacting the TCR binding platform can detect cognate CD8(+) T cells bearing low affinity TCRs that are not visible with the corresponding wildtype pHLA A*0201 multimeric complexes. Mechanistically, this effect is mediated by a disproportionate enhancement of the TCR/pMHCl association rate. In direct ex vivo applications, these coreceptor-enhanced multimers exhibit faithful cognate binding properties; concomitant increases in background staining within the non-cognate CD8(+) T cell population can be resolved phenotypically using polychromatic flow cytometry as a mixture of naive and memory cells. These findings provide the first validation of a novel approach to the physical detection of low avidity antigen-specific CD8(+) T cell populations; such coreceptor-enhanced multimeric reagents are likely to be useful in a multitude of settings for the detection of auto-immune, tumor-specific and cross-reactive CD8(+) T cells. (C) 2008 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available