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.

据信质膜在 成功建立了血源转移。 膜熊 （1）破坏地下膜屏障所需的酶， （2）\同质和异型细胞关联的受体以及 （3）\抗原，该抗原具有特异性和非特异性免疫反应性。 我 已经使用非溶解液与1-丁醇提取来增强发生率 实验诱导的肺转移。 丁醇增加 低转移性或高转移的转移潜力 源自化学诱导的纤维肉瘤，MCA-F或 自发黑色素瘤，B16。 将提取细胞在1至10 粗丁醇提取物（CBE）的微克重构转移 未提取控件的表型。 这项研究将隔离 并表征分子在细胞上出现的分子 表面，降低肿瘤细胞形成血源的电势 肺转移。 CBE中的重构活动将是 孤立的，结合制剂的等电聚焦，凝胶 过滤和凝集素亲和色谱和高压液体 色谱法。 部分纯化的重构活动 准备工作将被定量评估为毫克的数量 减少实验诱导的发生率所需的蛋白质 肺转移量增加了50％。 靶细胞的丁醇提取，但不是 效应细胞，降低天然细胞介导的细胞毒性百分比 （NCMC）在4小时的51 cr释放测定中。 原油和部分纯化 提取物将以可溶性形式使用或与目标相关联 细胞确定膜部分是否负责 血源转移酶的调节作为目标结构 NCMC。 静脉内使用CBE的合成宿主免疫 挑战显着增加了实验诱导的发生率 肺转移，这种作用可以通过脾 细胞。 有趣的是，可以在脾细胞的转移之前进行转移 或在挑战之后，表明CBE引起了免疫反应 特异性增强了肺部肿瘤的生长。 决赛 该项目的目的是描述细胞免疫反应 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 研究参与血源转移的可能机制， 肿瘤细胞表面结构在转移和 抗原刺激对转移最终发展的影响。