title: CC and CXC chemokine receptors mediate migration, proliferation, and matrix metalloproteinase production by fibroblast-like synoviocytes from rheumatoid arthritis patients.
abstract: OBJECTIVE
  To explore the potential involvement of the chemokine system in synoviocyte-mediated tissue destruction in rheumatoid arthritis (RA), we studied the expression profile of chemokine receptors and their function in the migration, proliferation, and matrix metalloproteinase (MMP) production of cultured fibroblast-like synoviocytes (FLS) from RA patients.
  
  
  METHODS
  The presence of CC and CXC chemokine receptors on cultured FLS was studied at the messenger RNA (mRNA) level by reverse transcriptase-polymerase chain reaction and at the cell surface expression level by flow cytometry. Variations in cytosolic calcium influx induced by chemokine stimulation were assessed by flow cytometry on Fura Red-preloaded FLS. Two-compartment transwell chambers were used for FLS chemotaxis assays. Cell growth was measured by a fluorescence-based proliferation assay. Gelatinase and collagenase activities were determined by a fibril degradation assay and zymography.
  
  
  RESULTS
  FLS constitutively expressed the receptors CCR2, CCR5, CXCR3, and CXCR4, both at the cell surface and mRNA levels, but failed to express CCR3 and CCR6. Significant intracytosolic calcium influx was observed on FLS challenged with monocyte chemotactic protein 1 (MCP-1), stromal cell-derived factor 1alpha (SDF-1alpha), and interferon-inducible protein 10 (IP-10). Stimulation with MCP-1, SDF-1alpha, IP-10, and monokine induced by interferon-gamma enhanced the migration and proliferation of FLS. These chemokines, in addition to RANTES, increased in a dose- and time-dependent manner the gelatinase and collagenase activities in cell-free supernatants of cultured FLS. Interestingly, the chemokine-mediated up-regulation of MMP activities was significantly abrogated by the presence of anti-interleukin-1beta, but not anti-tumor necrosis factor alpha, blocking antibodies.
  
  
  CONCLUSIONS
  These data suggest that through modulation of the migration, proliferation, and MMP production by FLS, the chemokine system may play a more direct role in the destructive phase of RA than is currently suspected, and thus emphasize the relevance of chemokines and their receptors as potential therapeutic targets in this disease.
authors.affiliation: Hospital Universitario de la Princesa, Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: Rosario
authors.initials: R
authors.lastname: García-Vicuña
authors.forename: Maria Victoria
authors.initials: MV
authors.lastname: Gómez-Gaviro
authors.forename: Maria Jesús
authors.initials: MJ
authors.lastname: Domínguez-Luis
authors.forename: Martina K
authors.initials: MK
authors.lastname: Pec
authors.forename: Isidoro
authors.initials: I
authors.lastname: González-Alvaro
authors.forename: Jose María
authors.initials: JM
authors.lastname: Alvaro-Gracia
authors.forename: Federico
authors.initials: F
authors.lastname: Díaz-González
mesh_terms: D001172:Arthritis, Rheumatoid / Q000378:metabolism* / Q000473:pathology; D002452:Cell Count; D002465:Cell Movement; D049109:Cell Proliferation; D018925:Chemokines / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000378:metabolism* / Q000473:pathology; D005434:Flow Cytometry; D006801:Humans; D019169:Jurkat Cells; D020782:Matrix Metalloproteinases / Q000096:biosynthesis*; D012333:RNA, Messenger / Q000378:metabolism; D019707:Receptors, Chemokine / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013583:Synovial Membrane / Q000378:metabolism* / Q000473:pathology
chemical_list: D018925:Chemokines; D012333:RNA, Messenger; D019707:Receptors, Chemokine; D020782:Matrix Metalloproteinases