|Budget Amount *help
¥2,200,000 (Direct Cost : ¥2,200,000)
Fiscal Year 1998 : ¥500,000 (Direct Cost : ¥500,000)
Fiscal Year 1997 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 1996 : ¥1,100,000 (Direct Cost : ¥1,100,000)
The concentration of Hepatocyte Growth Factor(HGF) significantly increases in synovial fluid obtained from RA patients and it correlated with serum C-reactive protein concentrations and the concentrations of IL-6 in synovial fluid, strongly suggesting the important role of HGF in systemic synovial inflammation in rheumatoid arthritis(RA).
Synovial fluid(SF) samples were obtained from patients with RA. SF cells were collected with centrifugation at 800 x g, and washed twice with phosphate buffered saline. Then the cells fractionated with Ficol-Isopaque gradient. Monocyte fraction and polymorphonuclear cell fraction were cultured at 5x10^6 cell/ml in RPMI1640 medium supplemented with 10% FCS for 1 to 3 days. Unfractionated synovial fluid cells were cultured for 3 hours, then culture supernatant containing non-adherent cells were moved to other culture plates. Adherent cells and non adherent cells were cultured separately for 1 to 3 days. The concentrations of HGF were measured by enzyme-l
inked immunosorbent assay. Time dependent secretion of HGF was observed only in polymorphonuclear cell fraction or adherent cells. 1,25-dihydroxyvitamin D_3 (1,25D_3) was added into the adherent cell culture at 10^<-11> to 10^<-6>M. HGF secretion was suppressed by 1,25D3 in dose dependent manner and the maximum suppression was observed yielding 60% at 10^<-7>M.
On the other hand, bone marrow(BM) specimens were obtained from 27 of RA patients, 1 of MCTD, 1 of SLE, 1 of PM/DM, one of Behcet's disease, 1 of FUO, whose mother is an RA patient, and healthy volunteers as normaal controls(CTL). 1 ml of BM specimens sampled from sternal bone and treated with EDTA and centrifuged. The concentrations of HGF and IL-6 in the supernatants were measured by enzyme-linked immunosorbent assay. HGF levels in BM of RA patients were significantly higher than that of CTL. HGF levels of RA were significantly correlated with serum C-reactive protein consentrations but not IL-6.
These results suggested that HGF in SF of RA positively correlated with RA activity is secreted by polymorphonuclear cell activated in bone marrow and migrated into SF, and active form of vitamin D may act immmunomodulater in RA. Less