CD14 contributes to the adherence of human monocytes to cytokine-stimulated endothelial cells.
Monocyte adherence to endothelial cells (EC) is selectively increased during inflammation. The mechanisms underlying monocyte-EC interaction indicated the involvement of surface-adhesion molecules on monocytes and EC. In earlier studies we noticed that the monocyte-specific mAb, designated mAb 63D3, in contrast to mAb against the beta 2-integrin molecules, inhibited the monocyte binding to monolayers of rIL-1 alpha-stimulated venous EC. The aim of the present study was to further characterize the Ag recognized by mAb 63D3 and to investigate the specific contribution of this Ag to the adherence of monocytes to cultured human macrovascular venous or arterial EC. Flow cytometric analysis demonstrated that the 63D3 Ag is expressed exclusively on the surface of peripheral blood monocytes. SDS-PAGE analysis of mAb 63D3 immunoprecipitates of 125I-labeled human monocyte surface proteins revealed that the target Ag for mAb 63D3 is a 52- to 55-kDa molecule identical to the myeloid differentiation protein CD14. Stimulation of EC with rIL-1 alpha or rTNF-alpha for 4 or 24 h or rIFN-gamma for 24 h increased (p less than 0.005) the number of monocytes bound to both types of EC. This cytokine-induced increase in monocyte adherence was significantly (p less than 0.0005) inhibited when the monocytes were coated with various mAb against CD14. The binding of monocytes to nonstimulated venous or arterial EC was not inhibited by anti-CD14 mAb. Our results lead to the conclusion that CD14 molecules, which on basis of their structure and m.w. are not related to the beta 2-integrin family of heterodimeric leukocyte adhesion molecules, participate in the binding of monocytes to cytokine-stimulated EC.
Beekhuizen
H
H
University Hospital, Department of Infectious Diseases, Leiden, The Netherlands.
Blokland
I
I
Corsèl-van Tilburg
A J
AJ
Koning
F
F
van Furth
R
R
D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000737:chemistry / Q000502:physiology*; D015214:Antigens, Differentiation, Myelomonocytic / Q000737:chemistry / Q000502:physiology*; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000502:physiology*; D016207:Cytokines / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology*; D005434:Flow Cytometry; D006098:Granulocytes / Q000166:cytology / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D018950:Lipopolysaccharide Receptors; D008214:Lymphocytes / Q000276:immunology; D008970:Molecular Weight; D009000:Monocytes / Q000166:cytology*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D015815:Cell Adhesion Molecules; D016207:Cytokines; D018950:Lipopolysaccharide Receptors