1988 Fiscal Year Final Research Report Summary
Studies of nephritogenic antigens in human renal basement membranes
| Project/Area Number |
61570478
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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 |
Pediatrics
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| Research Institution | Kinki University School of Medicine |
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Principal Investigator |
YOSHIOKA Kazuo Department of Pediatrics, Kinki University School of Medicine, Lecturer, 医学部小児科, 講師 (60111035)
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| Co-Investigator(Kenkyū-buntansha) |
ISEKI Takahiro Department of Pediatrics, Kinki University School of Medicine, fellow, 医学部小児科, 助手 (30176250)
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| Project Period (FY) |
1986 – 1988
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| Keywords | Glomerular basement membrane / Tubular basement membrane / Alveolar basement membrane / Anti-basement membrane antibody / Pulmonary hemorrhage / レクチン |
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| Research Abstract |
1. Analysis of nephritogenic antigens in human alveolar basement membrane (ABM). Goodpastue (DP) antigens, protein components reactive with human anti-glomerular basement membrane (GBM), were identified in human ABM using an enzyme-linked immunoassay, Western blotting and immunoprecipitation. Very cationic 22-28 kD and 40-48 kD components were detected by blot analysis combined with two-deimensional gel electrophoresis. Inhibition ELISA suggested anti-GBM antibodies recognized the same epitops(s) on the GBM and GBM. Heterogeneity of GP antigens in ABM was demonstrated by blotting: monomeric antigens were absent or minimal in ABM of three out of 10 normal individuals. In immunoprecipitation, anti-GBM antisera from patients with pulmonary hemorrhage precipitated GP antigens, byt those from patients without it did not. These data may explain why only some patients with anti-GBM nephritis have lung involvement.
2. Analysis of a target antigen in human anti-tubular basement membrane (TBM) nephritis. Collagenase digestion released antigenic moieties from normal human TBM. Anti-TBM antisera from four patients with anti-TBM nephritis showed on immunoprecipitation 54 kD and 48 kD with pI 7-8 and 6.5-7 antigenic components. Peincubation of normal kidney sections with RCA I, specific to galactose, inhibited the binding of anti-TBM antibody to TBM, suggesting the target antigen is a galactose containing glycoprotein. 54 kD antigen was isolated and purified using gel chromatography, ion exchange column, RCA I affinity column and reverse phase chromatography.
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