Monocyte adhesion to mesangial matrix modulates cytokine and metalloproteinase production

Abstract

Background. Monocytes migrate into the glomerular mesangium during acute inflammatory renal disease, differentiate into macrophages, and may play a key role in the development and progression of glomerular scarring. Treatment strategies that inhibit monocyte infiltration ameliorate glomerular injury in animal models. Mesangial matrix contains several potential monocyte-binding domains that may contribute to monocyte entrapment and modulate cell activation. Methods. Adhesion of peripheral blood-derived monocytes to matrix synthesized by human mesangial cells and to individual matrix proteins was assessed by colorimetry of nuclear staining with crystal violet. Monoclonal antibodies were used to identify the cell-surface integrins and matrix ligands involved. Monocyte proliferation was assessed by 3H-thymidine incorporation and cytokine production using enzyme-linked immunosorbent assay (ELISA). Secretion of metalloproteinases and their inhibitors was determined by zymography and ELISA, respectively. Results. Monocytes bound to matrix synthesized by mesangial cells. Prestimulation of mesangial cells with tumor necrosis factor-α (TNF-α) and transforming growth factor-β (TGF-β) enhanced matrix fibronectin content (P < 0.001) and monocyte binding (P < 0.001). Blocking antibodies to fibronectin, as well as to the integrins very late antigen-4 (VLA-4) and VLA-5, reduced monocyte adhesion to mesangial matrix by approximately 50%. Incubation of monocytes with matrix, fibronectin, laminin and collagen IV enhanced production of interleukin-1β (IL-1β), interleukin-6 (IL-6), TNF-α and metalloproteinase-9 (MMP-9) when compared to cells incubated in plastic wells. However, there was no apparent difference in proliferation rate and no change in production of metalloproteinase inhibitors. Conclusion. Monocyte activation within the glomerulus may be mediated by binding to mesangial matrix components, particularly fibronectin. Matrix-mediated activation enhances production of inflammatory cytokines and matrix-degrading enzymes.