© 1995 American Heart Association, Inc.
Articles |
Induction of Monocyte Chemoattractant Protein-1 Synthesis in Human Monocytes During Transendothelial Migration In Vitro
From the Departments of Cardiology (M.T., U.I., K.S.), Clinical Immunology (J.I.M., S.K.), and Medical Biology and Parasitology (T.K.) and the Division of Hemopoiesis (S.I.K.), Institute of Hematology, Jichi Medical School, Tochigi, and the Second Department of Internal Medicine (Y.I.T.), University of Tohoku, Miyagi, Japan.
Correspondence to Jun-Ichi Masuyama, MD, Department of Clinical Immunology, Jichi Medical School, Minamikawachi-machi, Tochigi 329-04, Japan.
Abstract Monocyte chemoattractant protein-1 (MCP-1, or
monocyte chemotactic and activating factor) plays important roles in
the recruitment of monocytes and thus in the development of
atherosclerosis. In this study, we determined whether MCP-1 synthesis
was induced by the cellular interaction between monocytes and
endothelial cells during the process of transendothelial migration. We
found that when human peripheral blood monocytes
(2.5x106 cells) and umbilical vein endothelial
cells (HUVECs; 5.0x105 cells) were cocultured for 5 hours,
7.9 ng/mL MCP-1 was secreted into the medium, whereas when the two were
cultured separately, MCP-1 levels were 1.0 and 0.9 ng/mL, respectively.
Furthermore, the use of interleukin-1ß (IL-1ß)–pretreated HUVECs
in cocultures induced twice the levels of MCP-1 as in unstimulated
HUVEC culture. Conditioned medium had transendothelial
chemotactic activity for monocytes, and this activity was completely
abolished by addition of anti–MCP-1 antibody. Although MCP-1 mRNA
levels were very low or undetectable in HUVECs or monocytes alone,
message could be detected after 2 hours of coculture in total mRNA
preparations from both monocytes and HUVECs. mRNA levels increased by 4
hours and had declined slightly by 24 hours. The rapid induction of
message suggests that cell contact between monocytes and HUVECs induces
the de novo synthesis of MCP-1 protein. Anti–interleukin (IL)-1
/ß
and anti–tumor necrosis factor- antibodies, or anti–lymphocyte
function–associated antigen-1 and very late antigen-4 antibodies, had
little or no inhibitory effects on MCP-1 secretion by cocultures.
Immunohistochemistry revealed that monocytes adherent to or having
migrated across unstimulated HUVEC monolayers as well as the
HUVECs themselves expressed MCP-1 protein. However, nonadherent
monocytes failed to express it. This finding suggests that the
monocyte–endothelial cell adhesive interaction results in an
MCP-1–inductive signal to each cell type. MCP-1 expression by migrated
monocytes may indicate that monocytes are primed to produce MCP-1
during transmigration and can secrete it in normal tissue in which
inflammatory cytokines that induce MCP-1 are otherwise absent.
Key Words: atherosclerosis • MCP-1 • monocyte • endothelium • migration
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