PMID- 28287509
OWN - NLM
STAT- MEDLINE
DCOM- 20180102
LR  - 20181202
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 120
DP  - 2017 Feb 6
TI  - Using X-ray Crystallography, Biophysics, and Functional Assays to Determine the 
      Mechanisms Governing T-cell Receptor Recognition of Cancer Antigens.
LID - 10.3791/54991 [doi]
LID - 54991
AB  - 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.
FAU - MacLachlan, Bruce J
AU  - MacLachlan BJ
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University.
FAU - Greenshields-Watson, Alexander
AU  - Greenshields-Watson A
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University.
FAU - Mason, Georgina H
AU  - Mason GH
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University.
FAU - Schauenburg, Andrea J
AU  - Schauenburg AJ
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University.
FAU - Bianchi, Valentina
AU  - Bianchi V
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University; Department of Oncology, University Hospital of Lausanne 
      (CHUV); Ludwig Insitutue for Cancer Research, Lausanne Branch, University of 
      Lausanne.
FAU - Rizkallah, Pierre J
AU  - Rizkallah PJ
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University.
FAU - Sewell, Andrew K
AU  - Sewell AK
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University.
FAU - Fuller, Anna
AU  - Fuller A
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University.
FAU - Cole, David K
AU  - Cole DK
AD  - Division of Infection and Immunity and Systems Immunity Research Institute, 
      Cardiff University; ColeDK@cardiff.ac.uk.
LA  - eng
GR  - Cancer Research UK/International
GR  - WT095767/Wellcome Trust/International
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20170206
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Antigens, Neoplasm)
RN  - 0 (Receptors, Antigen, T-Cell)
SB  - IM
MH  - Antigens, Neoplasm/*analysis
MH  - Biophysics/*methods
MH  - Crystallography, X-Ray/*methods
MH  - Humans
MH  - *Immunity, Cellular
MH  - Neoplasms/diagnosis/*immunology
MH  - Receptors, Antigen, T-Cell/*immunology
PMC - PMC5408581
EDAT- 2017/03/14 06:00
MHDA- 2018/01/03 06:00
PMCR- 2017/02/06
CRDT- 2017/03/14 06:00
PHST- 2017/03/14 06:00 [entrez]
PHST- 2017/03/14 06:00 [pubmed]
PHST- 2018/01/03 06:00 [medline]
PHST- 2017/02/06 00:00 [pmc-release]
AID - 54991 [pii]
AID - 10.3791/54991 [doi]
PST - epublish
SO  - J Vis Exp. 2017 Feb 6;(120):54991. doi: 10.3791/54991.