CCR7-mediated physiological lymphocyte homing involves activation of a tyrosine kinase pathway

Homing of blood-borne lymphocytes to peripheral lymph nodes (PLNs) Is a multistep process dependent on the sequential engagement of L-selectin, which mediates lymphocyte rolling along the luminal surface of high endothelial venules (HEVs), followed by activation of lymphocyte Integrins and transmigration through HEVs. Within lymphold tissue, B and T lymphocytes then migrate toward specific microenvironments such as B-cell follicles and the paracortex, respectively. The lymphocyte-expressed chemokine receptor CCR7 is playing an Important role during this process, as its HEV-presented ligands CCL19 and CCL21 can trigger rapid integrin activation under flow in addition to inducing a chemotactic response, which may participate in transmigration and/or interstitial migration. Here, we report that Tyrphostin (Tyr) AG490, a pharmacological inhibitor of Janus family tyrosine kinases (Jaks), blocked the chemotactic response of primary mouse lymphocytes to CCL19 and CCL21 in a dose-dependent manner. Furthermore, Tyr AG490 inhibited rapid CCL21-mediated up-regulation of alpha4 and beta2 integrin adhesiveness in static adhesion assays and under physiological flow, whereas adhesion induced by phorbol myristate acetate remained unaltered. Using intravital microscopy of subiliac PLNs in mice, we found that adoptively transferred Tyr AG490-treated lymphocytes adhered significantly less in HEVs compared with control cells, although L-selectin-mediated rolling was similar in both samples. Finally, we observed rapid Jak2 phosphorylation in CCL21-stimulated primary mouse lymphocytes. Thus, our study suggests a role for Jak tyrosine kinases during CCR7-mediated lymphocyte recirculation.