Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens.

Details

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State: Public
Version: Final published version
Serval ID
serval:BIB_B37051F6DB1F
Type
Article: article from journal or magazin.
Publication sub-type
Review (review): journal as complete as possible of one specific subject, written based on exhaustive analyses from published work.
Collection
Publications
Institution
Title
Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens.
Journal
Journal of visualized experiments
Author(s)
MacLachlan B.J., Greenshields-Watson A., Mason G.H., Schauenburg A.J., Bianchi V., Rizkallah P.J., Sewell A.K., Fuller A., Cole D.K.
ISSN
1940-087X (Electronic)
ISSN-L
1940-087X
Publication state
Published
Issued date
06/02/2017
Peer-reviewed
Oui
Number
120
Pages
0
Language
english
Notes
Publication types: Journal Article ; Video-Audio Media
Publication Status: epublish
Abstract
Human CD8+ cytotoxic T lymphocytes (CTLs) are known to play an important role in tumor control. In order to carry out this function, the cell surface-expressed T-cell receptor (TCR) must functionally recognize human leukocyte antigen (HLA)-restricted tumor-derived peptides (pHLA). However, we and others have shown that most TCRs bind sub-optimally to tumor antigens. Uncovering the molecular mechanisms that define this poor recognition could aid in the development of new targeted therapies that circumnavigate these shortcomings. Indeed, present therapies that lack this molecular understanding have not been universally effective. Here, we describe methods that we commonly employ in the laboratory to determine how the nature of the interaction between TCRs and pHLA governs T-cell functionality. These methods include the generation of soluble TCRs and pHLA and the use of these reagents for X-ray crystallography, biophysical analysis, and antigen-specific T-cell staining with pHLA multimers. Using these approaches and guided by structural analysis, it is possible to modify the interaction between TCRs and pHLA and to then test how these modifications impact T-cell antigen recognition. These findings have already helped to clarify the mechanism of T-cell recognition of a number of cancer antigens and could direct the development of altered peptides and modified TCRs for new cancer therapies.

Keywords
Antigens, Neoplasm/analysis, Biophysics/methods, Crystallography, X-Ray/methods, Humans, Immunity, Cellular, Neoplasms/diagnosis, Neoplasms/immunology, Receptors, Antigen, T-Cell/immunology
Pubmed
Web of science
Open Access
Yes
Create date
21/03/2017 18:14
Last modification date
20/08/2019 15:22
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