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PMID: 8104889 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Endotoxin activation of endothelium for polymorphonuclear leucocyte transendothelial migration and modulation by interferon-gamma.

Immunology ·Vol. 79 ·No. 4 ·1993-08-00 ·Pages 600-7

Issekutz AC, Lopes N

Abstract

Endotoxin [lipopolysaccharide (LPS)] is a potent inflammatory stimulus and can activate human umbilical vein endothelium (HUVE) for leucocyte adhesiveness and transendothelial migration. Here we investigated the role of HUVE-secreted cytokines in this process. When HUVE monolayers were grown on filters and preincubated for 3 hr with LPS, 51Cr-labelled polymorphonuclear leucocytes (PMNL) migrated across the HUVE in a dose- and time-dependent manner. Maximal PMNL transmigration with LPS (1 ng/ml) was 26 +/- 3% of added PMNL in 75 min. Neutralizing antibodies to interleukin-1 alpha (IL-1 alpha) and IL-1 beta, tumour necrosis factor-alpha (TNF-alpha), IL-8 or recombinant IL-1 receptor antagonist had no effect on the activation by LPS of the HUVE for supporting migration of PMNL. The HUVE 'activated state' declined with prolonged (22 hr) exposure to LPS, as reflected by a decrease in PMNL transendothelial migration to 5.5 +/- 1% and in the expression of the endothelial cell adhesion molecule, E-selectin, as compared to stimulation with LPS for 3 hr. However, simultaneous exposure to interferon-gamma (IFN-gamma) (200 IU/ml) and LPS maintained maximal PMNL transendothelial migration (28 +/- 4%) for at least 24 hr, prolonged E-selectin expression by HUVE and superinduced intracellular adhesion molecule-1 (ICAM-1) expression. The PMNL transendothelial migration was blocked by > 90% by monoclonal antibody (mAb) to CD18 with either 3 hr of LPS or 22 hr LPS + IFN-gamma stimulation. Migration was partially inhibited by mAb to E-selectin (30-40%) or to ICAM-1 (35-45%) and by a combination of both reagents (50-60%) under both stimulation conditions. Thus, LPS activation of HUVE for PMNL transendothelial migration: (a) does not require secretion of IL-1, TNF-alpha or IL-8 by the endothelium, (b) IFN-gamma enhances and prolongs endothelial activation by LPS and may increase leucocyte infiltration in LPS or bacterial inflammatory reactions, and (c) CD18-dependent mechanisms are equally important for PMNL transendothelial migration under both acute (3 hr) and prolonged (22 hr LPS + IFN-gamma) activation of endothelium.

MeSH Terms
Antibodies, Monoclonal/immunology Antigens, CD/immunology Cell Adhesion Cell Adhesion Molecules/immunology Cell Movement/immunology Cytokines/immunology Endothelium/immunology Endotoxins/immunology Humans Intercellular Adhesion Molecule-1 Interferon-gamma/immunology Lipopolysaccharides/immunology Neutrophils/immunology Time Factors Umbilical Veins/immunology
Chemicals
Antibodies, Monoclonal Antigens, CD Cell Adhesion Molecules Cytokines Endotoxins Lipopolysaccharides Intercellular Adhesion Molecule-1 Interferon-gamma
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Issekutz A C
Department of Pediatrics and Microbiology, Immunology, Dalhousie University, Halifax, NS, Canada.
Lopes N
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Article Info
Journal
Immunology
Abbr.
Immunology
ISSN
0019-2805
Published
1993-08-00
Pages
600-7
Language
English
Region
England
NLM ID
0374672
PMCID
PMC1421925
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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