Project/Area Number |
60480328
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Research Category |
Grant-in-Aid for General Scientific Research (B)
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Allocation Type | Single-year Grants |
Research Field |
Cerebral neurosurgery
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Research Institution | Kyushu University |
Principal Investigator |
FUKUI Masashi Professor and Chairman, Department of Neurosurgery, Faculty of Medicine, 医学部, 助教授 (10038713)
|
Project Period (FY) |
1985 – 1986
|
Project Status |
Completed (Fiscal Year 1986)
|
Budget Amount *help |
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1986: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1985: ¥4,400,000 (Direct Cost: ¥4,400,000)
|
Keywords | Neuroectodermal tumor / ganglioside / monoclonal antibody / blood brain barrier / BrdU |
Research Abstract |
The research project is to produce monoclonal antibody against ganglioside <GD_2> and to develop new diagnostic and therapeutic methods for brain tumors. Ganglioside <GD_2> is an oncofetal antigen expressed by fetal brain and neuroectodermal tumors. Bovine brain ganglioside <GD_(1b)> was extracted and converted to <GD_2> with bovine liver <beta> -galactosidase. <GD_2> was purified with Sephadex G-25 column chromatography. The purified <GD_2> was emulsified in complete Freund's adjuvant and chicken egg albumin. The antigen was weekly injected to mice intracutaneousely and intraperitoneally for a month. Any antibody reacting to <GD_2> was not detected in the sera of immunized mice. The spleen cells of immunized mice were fused to mouse myeloma cells(NS-1). The hybrid cells were selected with hypox-anthine-aminopterin-thymidine(HAT) media. Colony forming cells were very few and antibody producing cells were unable to obtain. On the other hand, many hybridoma cell lines were obtained from
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the hybrid cells between NS-1 and spleen cells of mice which were immunized with KNS-42 human glioma cell line. One of the hybridoma cell lines produces antibody reacting to cell surface proteins ( molecular weight: 65000 and 66000). The surface proteins localised in filopodea, ruffling membrane and adhesion plaque of cultured human glioma cells. Immunohistochemical studies on normal human brain and glioma revealed that the antibody specifically reacted to astrocytes and benign astrocytoma cells. Neurons and other mesenchymal cells were absolutely unstained. The functional roles of surface proteins are now studying. The proliferative activities analysed with bromodeoxyuridine(BrdU) uptake study revealed that the cases with high uptake of BrdU were poor outcome. The studies on blood brain barrier(BBB) disruption with mannitol revealed that the barriers of normal brain were more heavily destructed than those of tumors. BBB opening with intracarotid mannitol infusion may be hazardous on normal brain of patient. We should to keep up the fight against brain tumors and establish the effective method of controlling tumors. Less
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