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Production of chemokines, interleukin-8 and monocyte chemoattractant
protein-1, during monocyte: endothelial cell interactions
NW Lukacs, RM Strieter, V Elner, HL Evanoff, MD Burdick and SL Kunkel
Department of Pathology, University of Michigan Medical School, Ann Arbor
48109-0602, USA.
The extravasation of leukocytes from the lumen of the vessel to a site of
inflammation requires specific binding events. The interaction of
leukocytes with endothelium, via specific receptors, may provide
intracellular signals that activate extravasating cells. In the present
study, we have investigated the production of chemokines, interleukin-8
(IL-8), and monocyte chemoattractant protein-1 (MCP-1) during monocyte:
endothelial cell interactions. Both unstimulated and interferon-gamma
(IFN-gamma)-prestimulated human umbilical vein endothelial cells (HUVEC)
produced low constitutive levels of IL-8 and MCP-1. The addition of
enriched monocytes with unstimulated HUVEC resulted in synergistic
increases in production of both IL-8 and MCP-1. Monocytes cultured with
IFN-gamma-preactivated HUVECs demonstrated little additional increase in
IL-8 and MCP-1 production in coculture assays compared with unstimulated
HUVEC. Northern blot analysis paralleled the protein data, demonstrating
upregulated expression of IL-8 and MCP-1 mRNA in stimulated and
unstimulated coculture assays. Culture of enriched monocytes and
endothelial cells in transwells demonstrated no increases in IL-8 or MCP-1,
indicating the necessity for cellular contact for chemokine production. In
previous investigations, we have demonstrated that increased
monocyte-derived MIP-1 alpha production was induced by intracellular
adhesion molecule-1 (ICAM-1) interactions on activated HUVECs. In contrast,
addition of anti-ICAM-1 monoclonal antibodies (MoAbs) did not diminish the
production of IL-8 and MCP-1 in the present study. Furthermore, neither
antibodies to IL-1 nor tumor necrosis factor (TNF) diminished the
production of either IL-8 or MCP- 1. However, when soluble matrix proteins
were added to the coculture to block cellular interactions, the chemokine
protein and mRNA levels were significantly decreased. IL-8 production was
decreased by both soluble collagen and fibronectin, whereas MCP-1 was
decreased by only soluble collagen, suggesting differential activation
pathways. These results indicate that IL-8 and MCP-1 production are
increased during monocyte and endothelial cell interactions in part due to
matrix protein binding mechanisms. This mechanism may serve a role in cell
activation, production of chemokines, as well as extravasation and
recruitment of additional leukocytes during inflammatory responses.
Volume 86,
Issue 7,
pp. 2767-2773,
10/01/1995
Copyright © 1995 by The American Society of Hematology
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