Korniejewska Anna, Watson Malcolm, Ward Stephen
Department of Pharmacy and Pharmacology, University of Bath, Claverton Down, Bath, Slough, UK.
Methods Mol Biol. 2010;616:125-47. doi: 10.1007/978-1-60761-461-6_9.
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.
趋化因子（趋化性细胞因子）超家族成员及其受体在稳态和炎症条件下免疫细胞的运输中起主要作用。趋化因子受体CXCR3主要表达于活化的T淋巴细胞上，并结合三种促炎的、干扰素-γ诱导的趋化因子：干扰素-γ诱导的单核因子（Mig/CXCL9）、干扰素-γ诱导蛋白-10（IP-10/CXCL10）和干扰素-γ诱导的T细胞α趋化因子（I-TAC/CXCL11）。CXCR3及其激动剂参与多种炎症病理过程，使得该受体成为新型抗炎药物设计的一个有吸引力的靶点。有趣的是，越来越多的证据表明存在至少两种CXCR3的新型变体，即CXCR3-B和CXCR3-alt，这给针对CXCR3的新型抗炎药物设计带来了挑战。在本章中，我们描述了人外周血来源的T淋巴细胞的采集、分离和活化，以及在mRNA和蛋白质水平检测CXCR3及其非典型变体表达的方法，还有探索T淋巴细胞对所有已知CXCR3激动剂的生化和功能反应的实验方案。
