Analysis of CXCR3 and atypical variant expression and signalling in human T lymphocytes.
Members of the chemokine (Chemotactic cytokines) superfamily and their receptors play a major role in trafficking of immune cells under homeostatic and inflammatory conditions. The chemokine receptor CXCR3 is expressed mainly on activated T lymphocytes and binds three pro-inflammatory, interferon-gamma-inducible chemokines: monokine induced by IFN-gamma (Mig/CXCL9), IFN-gamma-induced protein-10 (IP-10/CXCL10) and IFN-gamma-inducible T-cell alpha-chemoattractant (I-TAC/CXCL11). CXCR3 and its agonists are involved in a variety of inflammatory pathologies, making this receptor an attractive target for the design of new anti-inflammatory drugs. Interestingly, a growing body of evidence suggests the existence of at least two novel variants of CXCR3, namely CXCR3-B and CXCR3-alt, which present challenges in the design of new anti-inflammatory drugs targeting CXCR3. In this chapter, we describe the collection, isolation and activation of human peripheral blood-derived T lymphocytes and methods to examine the expression of CXCR3 and its atypical variants at both mRNA and protein levels, as well as protocols for exploring the biochemical and functional responses of T lymphocytes to all known CXCR3 agonists.
Korniejewska
Anna
A
Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath, Slough, UK.
Watson
Malcolm
M
Ward
Stephen
S
D001681:Biological Assay / Q000379:methods*; D002465:Cell Movement; D049109:Cell Proliferation; D002469:Cell Separation; D004705:Endocytosis; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D054367:Receptors, CXCR3 / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D048348:Reverse Transcription / Q000235:genetics; D038601:Ribosomal Protein S6 / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*
C515187:CXCR3 protein, human; D020033:Protein Isoforms; D012333:RNA, Messenger; D054367:Receptors, CXCR3; D038601:Ribosomal Protein S6; D012313:RNA