Polymorphonuclear leucocyte migration through human dermal fibroblast monolayers is dependent on both β₂-integrin (CD11/CD18) and β₁-integrin (CD29) mechanisms

Accumulation of leucocytes in inflammation involves their migration through vascular endothelium and then in the connective tissue. We investigated human polymorphonuclear leucocyte (PMNL) migration through a biological barrier of human dermal fibroblasts grown on microporous filters, as a model of PMNL migration in the connective tissue. PMNL did not migrate through a fibroblast monolayer unless a chemotactic factor, e.g. C5a, interleukin-8 (IL-8) or zymosan-activated plasma (ZAP; C5a(desArg)), was added. This migration was partially inhibited (35-70%, depending on the stimulus) by treatment of PMNL with monoclonal antibody (mAb) to CD18 (β₂-integrins). Most of the CD18-independent migration was inhibited by mAb to β₁-integrins (CD29). Inhibition by mAb to β₁ was observed when the PMNL, but not the fibroblasts, were treated with mAb. The role of β₁-integrins in PMNL transfibroblast migration was detectable only when the function of the CD11-CD18 complex was blocked, because mAb to β₁-integrin alone had no significant effect on PMNL migration. Migration induced by C5a was more CD18-independent compared to IL-8 or C5a(desArg). The CD18-independent migration was also inhibited by mAb to the β₁-integrin subunits α₅ (of very late antigens-5; VLA-5) and α₆ (of VLA-6). Treatment of the fibroblasts (4 hr) with tumour necrosis factor-α (TNF-α) or IL-1α enhanced C5a-induced PMNL transfibroblast migration and increased the proportion of migration utilizing the CD11-CD18 mechanism. However, TNF-α treatment had no effect on the degree of β₁-integrin-dependent migration. These findings suggest that in response to the chemotactic factors C5a, IL-8 and C5a(desArg), PMNL migration in the connective tissue is mediated by both CD11-CD18 (β₂) and β₁-integrins on the PMNL. The VLA-5 and VLA-6 members of β₁-integrins are involved in this process. This is in contrast to PMNL migration across endothelium in this system, which is virtually all CD18 dependent with no significant role for β₁-integrins.