Integrin stimulation regulates polymorphonuclear leukocytes inflammatory cytokine expression

Objective The purpose of these studies is to investigate the role of integrin binding on the regulation of polymorphonuclear leukocyte (PMN) cytokine receptor expression. Summary Background Data Current knowledge in this area revolves around the ability of beta 2 integrins to mediate PMN adherence and chemotaxis. The role of alpha(1-6)/beta(1) integrins in regulating cytokine receptor expression has not been probed. Methods Purified human PMN were adhered on plastic, fibronectin, or laminin-coated surfaces followed by the addition of iodine 125 (I-125) monoclonal antibodies (mAbs) directed against tumor necrosis factor-alpha R (TNF-alpha R) p60, p80, or interleukin-1 beta R (IL-1 beta R) types I, II. Receptor expression was calculated based on the counts per minute (cpm) bound. The role of individual beta 1 integrins was assessed using mAbs directed against the alpha(1-6) subunit of the beta 1 complex, and integrin cross-linkage was achieved using secondary goat antimouse F(ab')(2) antibodies. Polymorphonuclear leukocytes were pretreated with herbimycin A to determine the role of protein tyrosine kinase in mediating the effect of the beta 1 integrins. Results Adherence of PMN to Ln decreased IL-1 beta types I, II receptor expression, whereas adherence to Fn increased TNF-alpha R p60 and p80 expression. Anti-VLA-5 (CD49e) but not anti-VLA-l through VLA-4 and VLA-6, blocked the effect of Fn on TNF-alpha receptors, whereas anti-VLA-6 but not anti-VLA-l through VLA-5 blocked the effect of Ln on IL-1 beta receptors. Modulation of IL-1 beta and TNF-alpha receptors by VLA-5 and VLA-6 required protein tyrosine kinase activation as herbimycin A (10 mu g/ml) blocked the affect of Fn and in. Changes in PMN cytokine receptor expression led to parallel changes in functional activity as assessed by the binding of I-125 IL-1 beta and TNF-alpha. Conclusions Integrin stimulation regulates the cell surface expression of PMN cytokine receptors. Ligation of CD49e upregulates TNF-alpha receptor expression, whereas binding of CD49f downregulates IL-1 beta receptor expression. Both processes are protein tyrosine kinase dependent. Changes in PMN cytokine receptor expression led to corresponding changes in functional activity. These results provide the first demonstration that chemotaxis of PMN into the interstitium provides a mechanism for ongoing participation in the local inflammatory response and is regulated by matrix protein integrin receptors.