1989 Fiscal Year Final Research Report Summary
Relationship between interstitial nephritis and tamm-Horsfall protein.
| Project/Area Number |
62480343
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Urology
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| Research Institution | Fujita Health University |
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Principal Investigator |
NAIDE Yorio Fujita Health University, School of Medicine, Department of Urology, Professor, 医学部・泌尿器科学, 教授 (40084561)
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| Co-Investigator(Kenkyū-buntansha) |
SHINODA Masayuki Fujita Health University, School of Medicine, Department of Urology, Assistant P, 医学部・泌尿器科学, 講師 (20138085)
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| Project Period (FY) |
1987 – 1989
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| Keywords | Tamm-Horsfall protein / ELISA / THP in serum |
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| Research Abstract |
Many investigators in Sweden and in the United States have studied to find a definite evidence of specific antibody against Tamm-Horsfall protein (THP) in serum of patients.
The first step of our study was also to find specific antibody against THP in serum of patients employing enzyme-linked immunosorberit assay technique(ELISA). This study was to confirm the results obtained by the study group in Goeteborg, Sweden, several years ago. We failed to confirm their data, however, and found that the technique used in their study missed an important step in a sequence of ELISA in which masking of test tube walls with serum albumin before addition of patient serum to avoid nonspecific binding of patients' serum globulins to the wall.
Therefore, high antibody titer against THP found in their study was simply looking at nonspecific binding of serum protein to the wall of test tubes.
Our next study was to find THP in patients' serum. We purified THP from human serum using affinity column-chromatography. Specific antibody against THP for preparing chromatography column was obtained by immunization of rabbit with THP from human urine.
Concentrated protein fraction obtained by repeated affinity chromatography was separated on SDS-PAGE and transblotted to nitrocellulose membrane. A band corresponding to THP was visualized by enzyme-antibody technique on the membrane using rabbit anti-human-THP antibody. Thus our result identified THP in human serum for the first time, although its presence has been suggested by radio-immuno assay technique in which antibody bound THP only could be detected.
In our further study, extraction of THP from rat urine is being attempted for immunizing rabbit with rat THP. Our aim is to observe THP histologically in experimentally infected rat kidney with varying underlying conditions or to determine whether anti THP antibody is produced in experimental infection.
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Research Products
(3 results)
