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Arthritis and Rheumatism Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD 20892
Monocytes/macrophages play a critical role in the initiation and
progression of a variety of glomerulonephritides. We sought to define
the interactions between physiologically activated human monocytes and
glomerular mesangial cells (MC) by employing a cell culture system that
permits the accurate assessment of the contribution of soluble factors
and cell-to-cell contact. Human peripheral blood monocytes, primed with
IFN-
and GM-CSF, were activated with CD40 ligand (CD40L) or TNF-
and cocultured with MC. CD40L-activated monocytes induced higher levels
of IL-6, monocyte chemoattractant protein-1 (MCP-1) and ICAM-1
synthesis by MC. Separation of CD40L-activated monocytes from MC by a
porous membrane decreased the mesangial synthesis of IL-6 by 80% and
ICAM-1 by 45%, but had no effect on MCP-1. Neutralizing Abs against
the ß2 integrins, LFA-1 and Mac-1, decreased IL-6
production by 40 and 50%, respectively. Ligation of mesangial surface
ICAM-1 directly enhanced IL-6, but not MCP-1, production. Simultaneous
neutralization of soluble TNF- and IL-1ß decreased MCP-1
production by 55% in membrane-separated cocultures of
MC/CD40L-activated monocytes. Paraformaldehyde-fixed CD40L-activated
monocytes (to preserve membrane integrity but prevent secretory
activity), cocultured with MC at various ratios, induced IL-6, MCP-1,
and ICAM-1 synthesis by MC. Plasma membrane preparations from activated
monocytes also induced mesangial IL-6 and MCP-1 synthesis. The addition
of plasma membrane enhanced TNF--induced mesangial IL-6 production
by 
4-fold. Together, these data suggest that the CD40/CD40L is
essential for optimal effector function of monocytes, that
CD40L-activated monocytes stimulate MC through both soluble factors and
cell-to-cell contact mediated pathways, and that both pathways are
essential for maximum stimulation of MC.
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