| Project/Area Number |
63570465
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Dermatology
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| Research Institution | Hirosaki University |
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Principal Investigator |
MITSUHASHI Yoshihiko Hirosaki University Hospital, Assistant Professor, 医学部附属病院, 講師 (30157557)
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| Co-Investigator(Kenkyū-buntansha) |
SAWAMURA Daisuke Hirosaki University School of Medicine, Assistant doctor, 医学部, 助手 (60196334)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1988: ¥600,000 (Direct Cost: ¥600,000)
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| Keywords | Type VII collagen / Anchoring fibril / Epidermolysis bullosa / Immunopathology / Electron microscopy / Bi-phase separated cell-culture system / Collagenase / Stromelysin / 二相分離培養法 / 7型コラーゲン / 皮膚 / 線維芽細胞 / インムノブロット |
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| Research Abstract |
(1) No studies on Japanese people concerning type VII collagen have been reported. We have examined healthy Japanese and Japanese DEB patients. We used two antibodies against two different epitopes of type VII collagen ; C-terminus and triple helical domain. We compared reactivities to these antibodies with expressions of collagenase and stromelysin mRNAs in fibroblasts from the same subject. Strong antigenicities against the both anti-type VII collagen antibodies were observed in 5 healthy volunteers and 1 dominant DEB patient. Four recessive DEB patients showed a reduced reactivity to the antibodies in a different manner in 4 each patient. The two examined epitopes showed a reduction in an almost the same degree in the each patient. These results may indicate that reduction of type VII collagen did not due to an enzymatic digestion, because proteolytic enzymes can digest the triple helical domain but not the C-terminus. On the other hand, expressions of collagenase and stromelysin mR
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NAs were considered to be an inverse correlation with degree of reactivity to the antibodies. This may support a possible participant of the collagenolytic enzymes in reduction of type VII collagen. Our examination was done on only a limited number of patients. Further studies are need to get any conclusion. (2) In order to find out in-vivo model for a study of DEB, we examined the wall of epidermal cysts. It was revealed that the epidermal cyst wall has an intact type VII collagen antigenicit and mature anchoring fibrils. Thus, epidermal cysts can not be substituted for the skin of DEB. (3) To know a process of anchoring fibril formation, we studied the dermal-epibolic junction which appears in an organ culture of normal skin. We observed that formation of anchoring fibrils can be devided into several steps. And it was suggested that type VII collagen was secreted beneath the basal lamina before the appearance of visible anchoring fibrils. (4) We tried to form anchoring fibrels in in-vitro experiments. We used a bi-phase separated cellculture systim in which keratinocytes and fibroblasts are simultaneously but separately cultured on the both sides of permeable collagen membrane. However, We could not find any basement membrane components at the basal aspect of basal cells. It is supposed together with the concomitant observation of the above described experiment (3) that pre-formed lamina densa and suitable dermal matrices in addition to a secretion of type VII collagen molecule are essential for a construction of mature anchoring fibrils. Less
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