Characterization of the migration of lung and blood T cells in response CXCL12 in a three-dimensional matrix

P>The ability of T cells to microlocalize within tissues, such as the lung, is crucial for immune surveillance and increased T-cell infiltration is a feature of many inflammatory lung conditions. T-cell migration has mainly been studied in two-dimensional assays. Using three-dimensional collagen gels to mimic the extracellular matrix of lung tissue, we have characterized the migration of T lymphocytes isolated from peripheral blood (PBT) and lung (LT) in response to interleukin-2 (IL-2) and CXCL12. Freshly isolated PBT and LT showed a low degree of migration (blood 4 center dot 0 +/- 1 center dot 3% and lung 4 center dot 1 +/- 1 center dot 7%). Twenty-four hours of culture increased the percentage of migrating PBT and LT (blood 17 center dot 5 +/- 2 center dot 9% and lung 17 center dot 7 +/- 3 center dot 8%). The IL-2 stimulation modestly increased migration of PBT after 6 days (32 center dot 3 +/- 6 center dot 0%), but had no effect on the migration of LT (25 center dot 5 +/- 3 center dot 2%). Twenty-four hours of stimulation with anti-CD3/CD28 caused a small but significant increase in the migration of PBT (to 36 center dot 4 +/- 5 center dot 8%). In a directional three-dimensional assay, CXCL12 failed to induce migration of fresh PBT or LT. Twenty-four hours of culture, which increased CXCR4 expression of PBT 3 center dot 6-fold, significantly increased the migration of PBT in response to CXCL12. Migration of PBT to CXCL12 was blocked by pertussis toxin, but not by the phosphoinositide 3-kinase inhibitor wortmannin. Twenty-four-hour cultured LT did not respond to CXCL12. CD3/CD28-stimulation inhibited CXCL12-mediated migration of PBT. These results suggest that the migration pattern of PBT is distinct from that of LT.