Detection of low avidity CD8(+) T cell populations with coreceptor-enhanced peptide-major histocompatibility complex class I tetramers
Melenhorst, Jan Joseph, 1965-, Scheinberg, Phillip, Chattopadhyay, Pratip K., Lissina, Anna, Gostick, Emma, Cole, David K., Wooldridge, L. (Linda), Berg, Hugo van den, 1968-, Bornstein, Ethan, Hensel, Nancy F., Douek, Daniel, Roederer, Mario, Sewell, Andrew K., Barrett, A. John and Price, D. A. (David A.). (2008) Detection of low avidity CD8(+) T cell populations with coreceptor-enhanced peptide-major histocompatibility complex class I tetramers. Journal of Immunological Methods, Vol.338 (No.1-2). pp. 31-39. ISSN 0022-1759Full text not available from this repository.
Official URL: http://dx.doi.org/10.1016/j.jim.2008.07.008
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.
|Item Type:||Journal Article|
|Subjects:||Q Science > QD Chemistry
Q Science > QH Natural history > QH301 Biology
Q Science > QR Microbiology > QR180 Immunology
|Divisions:||Faculty of Science > Mathematics
Faculty of Science > Centre for Systems Biology
|Library of Congress Subject Headings (LCSH):||Major histocompatibility complex, Flow cytometry, T cells|
|Journal or Publication Title:||Journal of Immunological Methods|
|Official Date:||30 September 2008|
|Number of Pages:||9|
|Page Range:||pp. 31-39|
|Access rights to Published version:||Restricted or Subscription Access|
|Funder:||National Institutes of Health (U.S.) (NIH), National Institute of Allergy and Infectious Diseases (U.S.). Vaccine Research Center (VRC), National Institute of Allergy and Infectious Diseases (U.S.) (NIAID), National Heart, Lung, and Blood Institute (NHLBI), Medical Research Council (Great Britain) (MRC), Wellcome Trust (London, England)|
Altman, J.D., et al., 1996. Phenotypic analysis of antigen-specific T lymphocytes.
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