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J Biol Chem, Vol. 274, Issue 2, 1050-1057, January 8, 1999
From Mechanisms regulating lipopolysaccharide
(LPS)-induced adherence to intercellular adhesion molecule (ICAM)-1
were examined using THP-1 cells transfected with CD14-cDNA
(THP-1wt). THP-1wt adherence to ICAM-1 was LPS dose-related,
time-dependent, and inhibited by antibodies to either CD14
or leukocyte function associated antigen (LFA)-1, but was independent
of any change in the number of surface expressed LFA-1 molecules. A
potential role for phosphatidylinositol (PI) 3-kinase (PI 3-kinase) in
LPS-induced adherence was examined using the PI 3-kinase inhibitors
LY294002 and Wortmannin. Both inhibitors selectively attenuated
LPS-induced, but not phorbol 12-myristate 13-acetate-induced adherence.
Inhibition by these agents was unrelated to any changes in either LPS
binding to or LFA-1 expression by THP-1wt cells. LPS-induced adherence
was also abrogated in U937 cells transfected with a dominant negative
mutant of of PI 3-kinase. Toxin B from Clostridium
difficile, an inhibitor of the Rho family of GTP-binding
proteins, abrogated both PI-3 kinase activation and adherence induced
by LPS. Cytohesin-1, a phosphatidylinositol
3,4,5-triphosphate-regulated adaptor molecule for LFA-1 activation, was
found to be expressed in THP-1wt cells. In addition, treatment of
THP-1wt with cytohesin-1 antisense attenuated LPS-induced adherence.
These findings suggest a model in which LPS induces adherence through a
process of "inside-out" signaling involving CD14, Rho, and PI
3-kinase. This converts low avidity LFA-1 into an active form capable
of increased binding to ICAM-1. This change in LFA-1 appears to be
cytohesin-1-dependent.
Monocyte Adherence Induced by Lipopolysaccharide Involves
CD14, LFA-1, and Cytohesin-1
REGULATION BY Rho AND PHOSPHATIDYLINOSITOL 3-KINASE
§,,,, and¶
Department of Medicine (Division of Infectious
Diseases) and the ¶ Department of Microbiology and Immunology, The
University of British Columbia, Faculties of Medicine and Science, The
Research Institute of the Vancouver Hospital and Health Sciences
Center, Vancouver, British Columbia V5Z 3J5, Canada and
§ The Laboratoire d'Immunologie, Faculté des Sciences
Dhar Mahraz, Université Mohamed Ben Abdallah, BP 1796, Atlas
Fés, Morocco
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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