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Diagnostic and therapeutic studies of ganglioside <GD_2> in human neuroectodermal tumors

神经节苷脂<GD_2>对人神经外胚层肿瘤的诊断和治疗研究

基本信息

批准号：
60480328
负责人：
FUKUI Masashi
金额：
$ 3.9万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60480328/
关键词：
Neuroectodermal tumor ganglioside monoclonal antibody blood brain barrier BrdU

项目摘要

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 … More 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

本研究项目旨在研制抗神经节苷脂单克隆抗体<GD_2>，为脑肿瘤的诊断和治疗提供新的方法。神经节苷脂<GD_2>是一种由胎儿脑和神经外胚层肿瘤表达的癌胚抗原。提取牛脑神经节苷脂（GD_（1b）），<GD_2>用牛肝<beta>半乳糖苷酶转化为GD_（1b）. <GD_2>经Sephadex G-25柱层析纯化。将纯化的蛋白<GD_2>在完全弗氏佐剂和鸡蛋白蛋白中乳化。每周皮内和腹腔内注射抗原给小鼠，持续一个月。免疫<GD_2>小鼠血清中未检测到任何与之反应的抗体。将免疫小鼠的脾细胞与小鼠骨髓瘤细胞（NS-1）融合。用次黄嘌呤-氨基蝶呤-胸苷（HAT）培养基筛选杂交细胞。集落形成细胞非常少，并且不能获得抗体产生细胞。另一方面，许多杂交瘤细胞系是从 ...更多信息 NS-1与KNS-42细胞免疫小鼠脾细胞的杂交细胞。杂交瘤细胞系之一产生与细胞表面蛋白（分子量：65000和66000）反应的抗体。培养的人脑胶质瘤细胞表面蛋白主要定位于丝状伪足、皱褶膜和粘附斑。对正常人脑和胶质瘤的免疫组化研究显示，该抗体特异性地与星形胶质细胞和良性星形细胞瘤细胞反应。神经元和其他间充质细胞完全未染色。表面蛋白的功能作用正在研究中，用溴脱氧尿苷（BrdU）摄取研究细胞增殖活性，发现BrdU高摄取者预后差，甘露醇对血脑屏障（BBB）的破坏研究表明正常脑屏障的破坏比肿瘤严重。颈动脉甘露醇灌注开放血脑屏障可能对患者的正常脑组织造成危害，应继续与脑肿瘤作斗争，建立有效的肿瘤控制方法。少