STIMULUS SPECIFICITY OF MATRIX METALLOPROTEINASE DEPENDENCE OF HUMAN T-CELL MIGRATION THROUGH A MODEL BASEMENT-MEMBRANE

Chemotaxis of human T lymphoblastoma cells of the Tsup-1 line, which m igrate similarly to blood T cells, through a layer of basement membran e-like Matrigel on a polycarbonate micropore filter was evoked by vaso active intestinal peptide (VIP; concentration for a maximal response, 10(-7) M), IL-2 (10(-9) M), IL-4 (10(-10) M), and the chemokines RANTE S (10(-10) M) and macrophage inflammatory protein-1 alpha (10(-10) M). Chemotactic concentrations of each factor increased Tsup-1 cell secre tion of matrix metalloproteinase-9 (MMP-9), with significant responses by 4 h for VIP, IL-2, and IL-4, but only after 24 h for macrophage in flammatory protein-alpha and RANTES, as quantified by Western blots an d zymography. H-3-Labeled type IV human collagen incorporated in the M atrigel layer was degraded by migrating Tsup-1 cells, as assessed by r elease of radioactive fragments of the collagen. The in situ degradati on of type IV collagen in Matrigel by migrating Tsup-1 cells was enhan ced most significantly by VIP, IL-2, and IL-4 after 4 h at concentrati ons that increased the secretion of MMP-9 optimally, but only after 24 h by macrophage inflammatory protein-1 alpha and RANTES. The specific MMP inhibitor GM6001 suppressed Tsup-1 cell MMP activity evoked by al l stimuli, as determined by zymography and in situ degradation of H-3- labeled type IV collagen. The chemotactic migration of Tsup-1 cells th rough Matrigel, but not through a filter alone, in response to optimal concentrations of VIP, IL-2, and IL-4, but not the chemokines, was in hibited by GM6001, with a concentration dependence similar to that for suppression of MMP activity. Thus elicitation of T cell chemotactic m igration through a model basement membrane by stimuli that increase MM P activity early in the response depends on degradation of matrix prot eins by MMP, whereas stimuli that recruit MMP late may rely on early a ctivation of other proteases.