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Blood, Vol. 93 No. 3 (February 1), 1999:
pp. 857-865
The MKK6/p38 Stress Kinase Cascade Is Critical for Tumor Necrosis
Factor- -Induced Expression of Monocyte-Chemoattractant
Protein-1 in Endothelial Cells
Matthias Goebeler,
Karin Kilian,
Reinhard Gillitzer,
Manfred Kunz,
Teizo Yoshimura,
Eva-B. Bröcker,
Ulf R. Rapp, and
Stephan Ludwig
From the Klinik und Poliklinik für Haut- und
Geschlechtskrankheiten and Institut für Medizinische
Strahlenkunde und Zellforschung (MSZ), Universität
Würzburg, Würzburg, Germany; and the Immunopathology
Section, Laboratory of Immunobiology, National Cancer
Institute-Frederick, Frederick, MD.
Monocyte chemoattractant protein-1 (MCP-1), a member of the C-C
subfamily of chemokines, is important for the local recruitment of
leukocytes to sites of inflammatory challenge. Here, we investigated endothelial signaling pathways involving members of the
mitogen-activated protein (MAP) kinase superfamily and studied their
role for MCP-1 expression in endothelium. We show that tumor necrosis
factor- (TNF- ), a potent inflammatory activator of endothelium,
leads to activation of MAP kinases ERK, p38, and JNK in human umbilical vein endothelial cells (HUVEC). Contribution of MAP kinase pathways to
TNF- -induced synthesis of endothelial MCP-1 was then studied by
pharmacologic inhibition and transient expression of dominant negative
or constitutively active kinase mutants using flow cytometry, Northern
blot, and luciferase reporter gene assays. Inhibition of Raf/MEK/ERK or
SEK/JNK pathways had no significant effect on MCP-1 levels, whereas
blocking the MKK6/p38 pathway by p38 inhibitors SB203580 or SB202190 or
by a dominant negative mutant of MKK6, the upstream activator of p38,
strongly inhibited TNF- -induced expression of MCP-1. Consistent
with that finding, expression of wild-type or constitutively active
MKK6 significantly enhanced the effect of limiting TNF-
concentrations on MCP-1 synthesis. These data suggest a crucial role
for the MKK6/p38 stress kinase cascade in TNF- -mediated endothelial
MCP-1 expression.

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