| Project/Area Number |
04670189
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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 |
Human pathology
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| Research Institution | Tokai University School of Medicine |
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Principal Investigator |
TSUTSUMI Yutaka Tokai University School of Medicine, Associate Professor, 医学部・病理学, 助教授 (80138643)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Keiichi Tokai University School of Medicine, Professor, 医学部・病理学, 教授 (00055865)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Helicobacter pylori / Enzyme-labeled antigen method / Chronic atrophic gastiritis / Intestinal metaplasia / Antibody-producing cells / Recombinant protein / Maltose-binding protein / Mucosal immunity / 慢性萎縮性胃炎 / 分泌型IgA |
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| Research Abstract |
It has been suggested that infection of Helicobacter pylori is pathogenic for chronic atrophic gastritis. However, no infection is demonstrated on the surface of intestinal metaplasia. The present study was aimed at identifying plasma cells producing antibodies specific for H.pylori in the mucosa with chronic atrophic gastritis by using "the enzyme-labeled antigen method". [Materials and Methods] Fresh gastric mucosa was sampled. We tried to prepare the labeled antigen in the following three approaches : A) Purification and biotinylation of urease, B) synthesis of biotinylated polypeptides from the known urease amino acid sequence. c) Production of recombinant protein of maltose-binding protein (MBP) and bacterial protein. [Results] Methods for histochemical detection of H.pylori was established. However, it was very difficult to prepare the labeled antigen. The methods A) and B) failed to be achieved. A preliminary study of "the enzyme-labeled antigen method" by using a biotinylated crude bacterial extract revealed no specific findings. Thus, with the method C), a recombinant fusion protein (M.W.60kD) was obtained by inserting a bacterial DNA fragment in the MBP-vector. The MBP-labeled protein showed a reactivity with the patient serum, but, regretably enough, it was of no use for detecting the specific antibody-producing cells in frozen sections. [Comments] The present study was unique in that the site of antibody production be visualized in the gastric mucosa by the novel technology. The preparation of the labeled antigen should be the starting point of study. We are now further trying to establish this approach for clarifying the pathogenesis of the bacteria in a chronic inflammatory process in the gastric mucosa.
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