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T-Cell Trafficking

Volume 616 of the series Methods in Molecular Biology pp 125-147

Date:

Analysis of CXCR3 and Atypical Variant Expression and Signalling in Human T Lymphocytes

  • Anna KorniejewskaAffiliated withDepartment of Pharmacy and Pharmacology, University of Bath
  • , Malcolm WatsonAffiliated withDepartment of Pharmacy and Pharmacology, University of Bath
  • , Stephen WardAffiliated withDepartment of Pharmacy and Pharmacology, University of Bath

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Abstract

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-γ-inducible chemokines: monokine induced by IFN-γ (Mig/CXCL9), IFN-γ-induced protein-10 (IP-10/CXCL10) and IFN-γ-inducible T-cell α-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.

Key words

T lymphocytes chemokines chemokine receptors CXCR3 chemotaxis PI3K