title: Expression of the duffy antigen/receptor for chemokines (DARC) by the inflamed synovial endothelium.
abstract: The expression of chemokine binding sites on the endothelial cells of venules in inflamed synovia was examined and whether the Duffy antigen/receptor for chemokines (DARC) was involved. In situ binding assays were performed to determine the expression of chemokine binding sites from rheumatoid (n = 10) and non-rheumatoid (n = 10) synovia. The expression of DARC protein and mRNA was examined by immunohistochemistry and northern blotting. The involvement of DARC in chemokine binding was studied by incubating sections with blocking antibodies to DARC (Fy3 and 6), to find out if these reduced 125I-IL-8 binding. Binding of radiolabelled chemokines IL-8, RANTES, MCP-1, but not MIP-1alpha, was found on venular endothelial cells in inflamed synovia from both rheumatoid and non-rheumatoid patients. Excess homologous unlabelled chemokine displaced binding and excess unlabelled RANTES could displace radiolabelled IL-8 binding. DARC protein expression was demonstrated on venular endothelial cells in all samples and DARC mRNA could be detected in extracts from synovia. There was downregulation of DARC protein and mRNA in rheumatoid samples. Binding of IL-8 to both rheumatoid and non-rheumatoid synovia was significantly reduced in the presence of anti-DARC Fy3 and Fy6 monoclonal antibodies. These findings show the expression of a multispecific chemokine binding site on the inflamed synovial endothelium, with evidence for involvement of DARC. This suggests a potential role for DARC in the inflammatory processes involved in synovitis.
authors.affiliation: Centre for Science and Technology in Medicine, Keele University at Robert Jones and Agnes Hunt Orthopaedic Hospital, Oswestry, UK. angela.patterson@rjahoh-tr.wmids.nhs.uk
authors.forename: Angela M
authors.initials: AM
authors.lastname: Patterson
authors.forename: Hilary
authors.initials: H
authors.lastname: Siddall
authors.forename: Giselle
authors.initials: G
authors.lastname: Chamberlain
authors.forename: Lucy
authors.initials: L
authors.lastname: Gardner
authors.forename: Jim
authors.initials: J
authors.lastname: Middleton
mesh_terms: D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal / Q000276:immunology; D000953:Antigens, Protozoan; D001172:Arthritis, Rheumatoid / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018925:Chemokines / Q000378:metabolism; D004375:Duffy Blood-Group System; D004730:Endothelium, Vascular / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008875:Middle Aged; D015800:Protozoan Proteins; D012333:RNA, Messenger / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D013583:Synovial Membrane / Q000098:blood supply / Q000378:metabolism; D013585:Synovitis / Q000378:metabolism*
chemical_list: C473745:ACKR1 protein, human; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D002352:Carrier Proteins; D018925:Chemokines; D004375:Duffy Blood-Group System; C066001:Duffy antigen binding protein, Plasmodium; D015800:Protozoan Proteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface