| Project/Area Number |
63480148
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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 |
寄生虫学(含医用動物学）
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| Research Institution | Asahikawa Medical College |
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Principal Investigator |
KATSUMI Haruhiko Asahikawa Medical College, Professor, 医学部, 教授 (90072868)
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| Co-Investigator(Kenkyū-buntansha) |
INAOKA Thoru Asahikawa Medical College, Assistant, 医学部, 助手 (00091562)
NAKAO Minorru Asahikawa Medical College, Assistant, 医学部, 助手 (70155670)
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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 |
¥5,800,000 (Direct Cost: ¥5,800,000)
Fiscal Year 1990: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1989: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1988: ¥3,400,000 (Direct Cost: ¥3,400,000)
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| Keywords | Alveolar Hydatid Disease / Monoclonal Antibody / Diagnosis / Treatment / モノクローナル抗体 / 診断と治療 |
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| Research Abstract |
Current biotechnology improved diagnostic and tools in parasitic diseases. In this study, murine monoclonal antibodies were produced to apply for the diagnosis and treatment of alveolar hydatid disease (AHD) caused by larval Echinococcus multilocularis.
Spleen cells from infected or immunized Balb/c mouse were fused with myeloma cells (P3X63-Ag8.653) by polyethylene glycol, and the hybridomas were grown in serum-free medium. ABC (avidin-biotin peroxidase complex) method using paraffin sections of alveolar hydatid was used as a screening assay. The hybridomas secreting antibodies specific for brood capsules, protoscolices or germinal layers were cloned by limiting dilution.
Fab fragments of the monoclonal antibody specific for germinal layers were radiolabeled with 131-I, and then injected into mice and rats bearing hepatic alveolar hydatids. However, effective images of the hepatic lesions were not obtained, probably because the antibody could not enter the inside of hydatids through the laminated layers. These results suggest that immunotoxins may be inapplicable for the treatment of AHD. In contrast, the immunohistological detection of germinal layers with monoclonal antibody was effective for evaluating the viable sites of hydatids in human hepatic lesions.
Antigenic characteristics in the Hokkido isolates of E. Multilocularis were examined by immunoblotting. The patient sera mainly reacted with two proteins of which molecular weights were 65 and 56-kilodalton (kDa). These antigens were different from the 54kDa antigens, designated "Em2" in the European isolate, and were nonreactive with sera of other parasitic infections. Accordingly, the production of monoclonal antibodies against these antigens is necessary to purify the immunodiagnostic antigens.
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