title: Anti-inflammatory effects of leflunomide on cultured synovial macrophages from patients with rheumatoid arthritis.
abstract: BACKGROUND
  Leflunomide and its active metabolite A77 1726 reversibly inhibits the enzyme dihydro-orotate dehydrogenase, the rate limiting step in de novo synthesis of pyrimidines and progression of the cell cycle in different cell lines, mainly activated T lymphocytes.
  
  
  OBJECTIVE
  To analyse in vitro the possible anti-inflammatory effects exerted by A77 1726, on cultured macrophages, obtained from the synovial tissues of patients with rheumatoid arthritis (RA).
  
  
  METHODS
  The effects of different doses of A77 1726 on intracytoplasmic expression and extracellular concentration of inflammatory cytokines (tumour necrosis factor alpha (TNFalpha), interleukin (IL) 1beta, IL6), as well as the influence on production and expression of intercellular adhesion molecule-1 (ICAM-1) and cyclo-oxygenase 2 (COX-2) by primary cultures of synovial macrophages from patients with RA, were evaluated by immunocytochemistry and western blot analysis. The observations were made at four and 24 hours.
  
  
  RESULTS
  A progressive and significant time and dose dependent decrease of the number of positive macrophages for intracellular TNFalpha and IL1beta, treated with different doses of A77 1726, was found in comparison with untreated cells. The extracellular concentration of TNFalpha was found to be significantly decreased in media containing cultured macrophages at 24 hours for all tested doses of A77 1726. At 24 hours, a significant time and dose dependent decrease of ICAM-1 and COX-2 expression by cultured macrophages after A77 1726 treatment was found.
  
  
  CONCLUSIONS
  In conclusion, the mechanism of antiproliferative activity exerted by leflunomide on activated T lymphocytes seems to be the same mechanism (alteration of the cell cycle progression) which interferes with the functions of other activated cells-namely, the monocytes/macrophages, which are strongly involved in the inflammatory reaction in RA synovial tissue. The positive clinical results seem to confirm that leflunomide exerts an anti-inflammatory action on phagocytic cells in short and long term treatment of RA.
authors.affiliation: Research Laboratory and Division of Rheumatology, Department of Internal Medicine University of Genova, Italy. mcutolo@unige.it
authors.forename: M
authors.initials: M
authors.lastname: Cutolo
authors.forename: A
authors.initials: A
authors.lastname: Sulli
authors.forename: P
authors.initials: P
authors.lastname: Ghiorzo
authors.forename: C
authors.initials: C
authors.lastname: Pizzorni
authors.forename: C
authors.initials: C
authors.lastname: Craviotto
authors.forename: B
authors.initials: B
authors.lastname: Villaggio
mesh_terms: D000814:Aniline Compounds / Q000494:pharmacology*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D018501:Antirheumatic Agents / Q000494:pharmacology; D001172:Arthritis, Rheumatoid / Q000473:pathology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003437:Crotonates; D051546:Cyclooxygenase 2; D016207:Cytokines / Q000096:biosynthesis; D003593:Cytoplasm / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D006885:Hydroxybutyrates / Q000494:pharmacology*; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D007527:Isoenzymes / Q000096:biosynthesis; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism; D008565:Membrane Proteins; D008875:Middle Aged; D009570:Nitriles; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D013583:Synovial Membrane / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D014052:Toluidines; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D000814:Aniline Compounds; D000894:Anti-Inflammatory Agents, Non-Steroidal; D018501:Antirheumatic Agents; D003437:Crotonates; D016207:Cytokines; D006885:Hydroxybutyrates; D007527:Isoenzymes; D008565:Membrane Proteins; D009570:Nitriles; D014052:Toluidines; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1; C527525:teriflunomide; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases