Differential influences of bucillamine and methotrexate on the generation of fibroblast-like cells from bone marrow CD34+ cells of rheumatoid arthritis patients.
We have recently demonstrated that bone marrow CD34+ cells from rheumatoid arthritis (RA) patients displayed abnormal capacities to respond to TNF-alpha and to differentiate into fibroblast-like cells producing MMP-1 (type B synoviocyte -like cells). The current study examined the effects of representative potent disease-modifying antirheumatic drugs, including bucillamine (BUC) and methotrexate (MTX) on the in vitro generation of fibroblast-like cells from RA bone marrow CD34+ cells. CD34+ cells purified from bone marrow specimens of 8 patients with active RA were cultured in the presence or absence of pharmacologically attainable concentrations of intramolecular disulfide form of bucillamine (BUC-ID, 3 microM), a major metabolite of BUC or MTX (20 nM). After incubation for 28 days, the generation of fibroblast-like cells was assessed under phase-contrast light microscopy and the concentrations of MMP-1 and VEGF in the culture supernatants were measured by ELISA. BUC-ID, but not MTX, significantly suppressed the generation of fibroblast-like cells from RA bone marrow CD34+ cells stimulated with SCF, GM-CSF and TNF-alpha (p=0.024 as determined by Wilcoxon signed rank test). Accordingly, BUC-ID, but not MTX, significantly suppressed the production of MMP-1 (p=0.017) and VEGF (p=0.017) by RA bone marrow CD34+ cells, without inhibition of beta2-microglobulin production. These results demonstrate that BUC-ID, but not MTX, is a potent inhibitor of differentiation of fibroblast-like cells from RA bone marrow CD34+ cells. Since MTX, but not BUC, has been previously shown to influence on type A synoviocytes, the data provide rationale of combination of BUC and MTX in the treatment of RA.
Hirohata
Shunsei
S
Kitasato University School of Medicine, Kanagawa 228-8555, Japan. shunsei_tenpoint@yahoo.co.jp
Yanagida
Tamiko
T
Tomita
Tetsuya
T
Yoshikawa
Hideki
H
D000328:Adult; D000368:Aged; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D018952:Antigens, CD34 / Q000378:metabolism*; D018501:Antirheumatic Agents / Q000494:pharmacology*; D001172:Arthritis, Rheumatoid / Q000276:immunology / Q000473:pathology*; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000276:immunology*; D003470:Culture Media; D003545:Cysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D005260:Female; D005347:Fibroblasts / Q000187:drug effects*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007085:Ilium / Q000166:cytology; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007202:Indicators and Reagents; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D008727:Methotrexate / Q000494:pharmacology*; D008875:Middle Aged; D013234:Stem Cells / Q000187:drug effects; D014746:Vimentin / Q000494:pharmacology
D000894:Anti-Inflammatory Agents, Non-Steroidal; D018952:Antigens, CD34; D018501:Antirheumatic Agents; D003470:Culture Media; D007166:Immunosuppressive Agents; D007202:Indicators and Reagents; D014746:Vimentin; D020781:Matrix Metalloproteinase 1; D003545:Cysteine; C026535:bucillamine; D008727:Methotrexate