1994 Fiscal Year Final Research Report Summary
Development of an early detection method and specific immunotherapy of ovarian cancers
| Project/Area Number |
05454439
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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 |
Obstetrics and gynecology
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| Research Institution | Chiba Univ. |
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Principal Investigator |
SEKIYA Souei Chiba Univ.Sch.Med.Prof., 医学部, 教授 (00092065)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Naotake Chiba Univ.Sch.Med.Assistant, 医学部・付属病院, 助手 (80236611)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Ovarian cancer / Monoclonal antibody / Immune complex / serum-free culture / Ascites |
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| Research Abstract |
To develop a new diagnostic method for ovarian cancers at an carly stage, we produced murine monoclonal antibodies (MoAbs) against ovarian cancer-derived molecules which are lmmunogenic in patients. Five different strategies of immunizing antigen/screening antigen combination were tried. In all strategies, immune complexes (ICs) purified from ascites of 30 patients with various ovarian cancers were used as immunizing antigens and/or screening antigens in ELISA.The purification of ICs includes ammonium sulphate precipitation. Sephacryl S300 gel filtration, and protein A affinity chromatography. As immunizing antigens, serum-free conditioned medium of a serous cystadenocarcinoma cell line (HOC/BR) or tumor tissue of ovarian cancers were also used. ICs purified from pooled sera of normal healthy controls were used as negative screening antigens. We screened more than 8,000 supernatants of hybridomas in to fusion experiments, and established 7 MoAbs which react with ascites LCs of ovarian cancers, but not with IgG or ICs of normal controls. Among them, 2 MoAbs (2Flt and 7E2 clones) can also detect circulating ICs in peripheral blood from patients with ovarian cancers by sandwich ELISA using protein A precoated micropiates. Conditioned medium of HOC/BR cell line inhibited the reactivity of 3 MoAbs (2Fll, 7E2, and H5 clones), indicating that these antibodies detect ovarian cancer-derived molecules involved in the patients' ICs. These molecules are, therefore, considered to express tumor-associated epitopes reacted by patients' IgG.
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