MEMBRANE ANTIGENS WHICH MEDIATE METASTATIC PHENOTYPE

介导转移表型的膜抗原

• 批准号: 3176572
• 负责人: STEPHEN J LE GRUE
• 金额: $ 9.19万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-03-01 至 1986-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3176572
• 关键词: affinity chromatography; autoradiography; butanols; cell mediated cytotoxicity; electrofocusing; gel filtration chromatography; gene expression; high performance liquid chromatography; lung neoplasms; membrane activity; metastasis; neoplasm /cancer genetics; neoplasm /cancer immunology; neoplastic cell; radiotracer; spleen; tumor antigens

The plasma membrane is believed to play an important role in the successful establishment of hematogenous metastases. The membrane bears (1)\enzymes required for destruction of basement membrane barriers, (2)\receptors for homotypic and heterotypic cell associations and (3)\antigens which direct specific and nonspecific immune reactivities. I have used noncytolytic extraction with 1-butanol to enhance the incidence of experimentally induced pulmonary metastases. Butanol increases the metastatic potential of sublines of either low or high metastatic phenotype derived from the chemically induced fibrosarcoma, MCA-F, or the spontaneous melanoma, B16. Brief incubation of extracted cells in 1 to 10 micrograms of crude butanol extract (CBE) reconstituted the metastatic phenotype to that of the unextracted controls. This study will isolate and characterize the molecules which, when they are borne on the cell surface, reduce the potential of neoplastic cells to form hematogenous pulmonary metastases. The reconstitutive activity in CBE will be isolated, using a combination of preparative isoelectric focusing, gel filtration and lectin affinity chromatography and high-pressure liquid chromatography. The reconstitutive activity in partially purified preparations will be quantitatively assessed as the milligram amount of protein necessary to reduce the incidence of experimentally induced pulmonary metastases by 50%. Butanol extraction of target cells, but not effector cells, reduces the percent natural cell-mediated cytotoxicity (NCMC) in a 4-hour 51Cr-release assay. Crude and partially purified extracts will be used either in soluble form or associated with target cells to determine whether the membrane moieties responsible for the modulation of hematogenous metastases serve as target structures for NCMC. Immunization of syngeneic hosts with CBE before intravenous challenge significantly increases the incidence of experimentally induced lung metastases, and this effect can be adoptively transferred with spleen cells. Interestingly, transfer of spleen cells can be made either before or after challenge, suggesting that CBE elicits an immune response which specifically potentiates the growth of tumor in the lungs. The final objective of this project is to delineate the cellular immune response to antigens in CBE and to characterize the antigens and immune cells participating. Thus, butanol extraction of metastatic cells will allow investigation of possible mechanisms involved in hematogenous metastasis, of the role of tumor cell surface structures in metastasis and of the effect of antigen stimulation on the eventual development of metastases.

质膜被认为在 成功建立血行转移。 膜熊 (1)\破坏基底膜屏障所需的酶， (2)\同型和异型细胞关联的受体和 (3)\指导特异性和非特异性免疫反应的抗原。 我 使用 1-丁醇进行非溶细胞提取以提高发生率 实验诱导的肺转移。 丁醇增加 低或高转移亚系的转移潜力 源自化学诱导的纤维肉瘤、MCA-F 或 自发性黑色素瘤，B16。 将提取的细胞短暂孵育 1 至 10 微克粗丁醇提取物 (CBE) 重构转移瘤 表型与未提取的对照的表型相同。 这项研究将隔离 并表征分子，当它们承载在细胞上时 表面，降低肿瘤细胞形成血细胞的潜力 肺转移。 CBE 的重组活动将是 分离，使用制备等电聚焦、凝胶的组合 过滤和凝集素亲和层析和高压液体 色谱法。 部分纯化的重构活性 制剂将被定量评估为毫克量 减少实验诱导的发生率所必需的蛋白质 肺转移50%。 丁醇提取靶细胞，但不 效应细胞，降低自然细胞介导的细胞毒性百分比 (NCMC) 在 4 小时 51 Cr 释放测定中。 粗品和部分纯化 提取物将以可溶形式或与目标结合使用 细胞以确定膜部分是否负责 血行转移的调节作为靶结构 NCMC。 静脉注射前用 CBE 对同基因宿主进行免疫 挑战显着增加了实验诱导的发生率 肺转移，这种效应可以通过脾脏过继转移 细胞。 有趣的是，脾细胞的转移可以在 或在攻击后，表明 CBE 引发了免疫反应， 特别增强肺部肿瘤的生长。 决赛 该项目的目标是描绘细胞免疫反应 CBE 中的抗原并表征抗原和免疫细胞 参与。 因此，转移细胞的丁醇提取将允许 研究血行转移的可能机制， 肿瘤细胞表面结构在转移中的作用以及 抗原刺激对转移最终发展的影响。