| Project/Area Number |
61570334
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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 |
Gastroenterology
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| Research Institution | Shinshu University School Of Medicine. |
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Principal Investigator |
KIYOSAWA Kendo Department of Internal Medicine, Shinshu University School of Medicine., 医学部・文部教官, 助教授 (30020829)
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| Co-Investigator(Kenkyū-buntansha) |
GIBO Yukio Department of Internal Medicine, Shinshu University School of Medicine., 医学部・文部教官, 助手 (40161616)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | proliferating cell nuclear antigen (PCNA) / anti-PCNA / antinuclear antibody / PCNA / 核抗原 / 肝癌 |
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| Research Abstract |
1. Sera from 160 patients with hepatocellular carcinoma(group I) were examined for anti-nuclear antibodies(ANA) indirect immunofluorescence using 3 types of cultured as substraye : human hepatoma cell(PLC/PRF/5), laryngeal carcinoma cell(HEp-2) and baby hamster kidney cell (BHK). Sixty six sera from patients with non-hepatic cancers (group II), 62 sera from patients with chronic liver disease (group III) and 235 sera of normal subjects (group IV) were tested as controls. The incidences of ANA which was detected in HEp-2 or pLC/PRF/5 but not in BHK were as follows, 36 patients(22.5%) in group I, 19 patients(27.5%) in group II, 12 patients (19.4%) in group III and only 1 case(0.4%) in group IV. Reciprocal titers of ANA that were positive against HEp-2 but negative against BHK in group I and II were higher than in group III and IV. Two Two sera of patients with hepatocellular carcinoma reacted with nuclei of HEp-2 and PLC/PRF/5 were investigated further for ANA by indirected immunofluores
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cence using other 4 types of transformed cells and 4 types of non-transformed cells. We observed that in these two sera reacted only with transformed cell differed from that in sera of patients with SLE. It was suggested that the ANA reacting only with nuclei of transformd cells was present in sera of patients with hepatocellular carcinoma. We need weather this tumor specific antibody cross react with antibody to proliferating cell nuclear antigen(PCNA).
2. Purification of PCNA was performed by affinity chromatography. Calf thymus was used as a source of antigen. IgG fraction was obtained from anti-PCNA positive lupus serum and absorbed other antibodies to extractable nuclear antigens by affinity column coupled with rabbit kidney extract. Anti-PCNA IgG antibody was conjugated with ligand gels to make PCNA affinity coloumn. Calf thymus acetone powder was homogenized with PBS and the supernatant was dialysed against water, then the precipitate was used as crude PCNA. The antigen was passed over the anti-PCNA coupled column and the eluted fraction was taken as PCNA. This purified antigen was a 29 kd in molecular weight andreacted only with anti-PCNA serum in both immunoblotting and enzyme immunoassay. Less
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