Production of human monoclonal antibody to OFA-1 and it's cytotoxicity to trophoblasts
| Project/Area Number |
62570745
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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 |
Obstetrics and gynecology
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| Research Institution | NIIGATA UNIVERSITY |
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Principal Investigator |
YUZAWA Hideo Niigata University School of Medicine, 医学部附属病院, 講師 (30143770)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAKUWA Koichi Niigata University School of Medicine, 医学部, 助手 (80187939)
KANAZAWA Koji Niigata University School of Medicine, 医学部, 助教授 (50092680)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1988: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1987: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | pregnancy / Ganglioside / GM2 / Oncofetal Antigen Immunogenic / ヒトモノクローナル抗体 / 妊娠免疫 / OFA-I / ガングリオシッド |
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| Research Abstract |
Biochemical analysis revealed that OFA-1 is ganglioside GM2 and GD2. In the present study, we tried to produce human monoclonal antibody against GM2 and examine cytotoxic activity of the human monoclonal antibody to trophoblasts. Peripheral Blood Lymphocyted (PBL) were obtained from pregnant woman whose anti-GM2 activity was highest. PBL were hybridized with KR-12 and/or immortalized by EBV several times, but specific clone was not established. There, for the purpose of specific B cell clone expanssion. PBL were stimulated in vitro by GM2 and then hybridized with KR-12. However, it had not been possible to produce the specific human monoclonal antibody. So, in this ^<51>Cr releasing assay, we used the pregnant woman's serum which is specific to GM2 by competitive inhibition assay instead of human monoclonal antibody. Cytotoxicity of this serum to melanoma cell line M-14( this line expressed GM2 on the cell surface) was observed strongly, otherwise cytotoxicity to trophoblasts was not obserbed. This results suggest that GM2 may not be expressed on the trophoblasts.
So, GM2 was investigated as to the tissue localization in pregnancy by Thin Layer Chromatogram (TLC)-Immunostaining method and Immuno-histological method. The results obtained were as follows:(1) A concentration of GM2 in pregnant women's sera was about 7 times greater than that of cord serum. (2) GM2 was betected clearly only in the becidual layer, but not detected on the trophoblasts and any other tissues.
These results suggested that a prigin of GM2 in pregnant women's sera was GM2 from decidua and anti-GM2 antibody was induced by this GM2.
On the contrary, anti-GM2 antibody did not have cytotoxic activity to trophoblasts. This is reasonable for a success of pregnancy.
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Report
(3 results)
Research Products
(5 results)
