The role of cytokines in the pathogenesis of fibrosis in systemic sclerosis (scleroderma)

Research Project

Project/Area Number	62570295
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	Legal medicine
Research Institution	Kitasato University
Principal Investigator	KONDO Hirobumi Kitasato Univ. Sch. of Med. associate prof., 医学部, 助教授 (70124922)
Co-Investigator(Kenkyū-buntansha)	KASHIWAZAKI Sadao Kitasato Univ. Sch. of Med. prof., 医学部, 教授 (20050429) TAKASHINA Naoya Kitasato Univ. Sch. of Med. lecturer, 医学部, 助手 (00187949) SUZUKI Takahiro Kitasato Univ. Sch. of Med. lecturer, 医学部, 助手 (00179227)
Project Period (FY)	1987 – 1989
Project Status	Completed (Fiscal Year 1989)
Budget Amount *help	¥1,500,000 (Direct Cost: ¥1,500,000) Fiscal Year 1989: ¥700,000 (Direct Cost: ¥700,000) Fiscal Year 1988: ¥800,000 (Direct Cost: ¥800,000)
Keywords	systemic sclerosis / scleroderma / fibrosis / cytokine / fibroblast proliferation / collagen secretion / IL-1 / 全身制硬化症 / TNF / 線維芽細胞 / 1Lー1 / IFNーγ / コラーゲン
Research Abstract	Systemic sclerosis (scleroderma) is characterized by diffuse accumulation of collagen and vascular change in the skin and a variety of internal organs. We planned to evaluate the role of cytokines in the pathogenesis of fibrosis of scleroderma. The supernates of LPS-stimulated and unstimulated plastic dish adherent mononuclear cells (monocute rich) from scleroderma patients and iaormal controls were used as monokine. The monokine supernates from scleroderma MNC resulted in greater proliferation of skin fibroblast detected by 3H-thymidine incorporation than those from normal controls. When the supernate was fractionated by HPLC gel filtration, the high proliferation activity was consisted in 22-35KD fraction (peak 24-26KD) of either scleroderma or normal supernate. The activity of the supernate was partially inhibited by antisera to IL-1 alpha, IL-1 beta and TNF alpha, which suggested the supernate included these cytokines. The supernate and the fractionated supernate included various concentrations of IL-1 alpha and IL-1 beta detected by ELISA. A tendency to an increase in the alpha / beta ratio of IL-1 was seen in the scleroderma supernate. Collagen secretion from fibroblasts in the culture supernate was determined by the method of Peterkofsky and Diegelmann. The monokine supernate of LPS-stimulated MNC from normal controls (10% concentration) inhibited the secretion of collagen ranged from 10 to 62%. The inhibitory activity of the supernate from scleroderma MNC was lower than that of normal MNC. The responses to serial concentration of rIL-1 alpha, rTNF alpha and rIFN-gamma were compared between scleroderma and normal fibroblasts. These was no significant difference between these fibroblasts in responses to these cytokines. These results indicate that cytokines including IL-1 and TNF may contribute to the increased collagen accumulation observed in scleroderma.