MEMBRANE ANTIGENS THAT MEDIATE METASTATIC PHENOTYPE

介导转移表型的膜抗原

• 批准号: 3176573
• 负责人: STEPHEN J LE GRUE
• 金额: $ 14.96万
• 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 1989-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3176573
• 关键词: affinity chromatography; autoradiography; basement membrane; butanols; cell mediated cytotoxicity; electrofocusing; gel filtration chromatography; gene expression; high performance liquid chromatography; ion exchange 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. The purpose of this study is to isolate and characterize cells surface moieties that influence metastatic phenotype, using a combination of butanol extraction, preparative isoelectric focusing, high performance gel permeation, ion exchange and hydrophobic interaction chromatography, and lectin affinity chromatography. The applicant has employed noncytolytic extraction using butanol to enhance the incidence of experimentally-induced pulmonary metastases. Extraction increases the metastatic potential of sublines of either low or high metastatic phenotype derived from the chemically induced fibrosarcoma, MCA-F, or spontaneous melanoma, B16. Brief incubation of extracted cells in crude butanol extracts (CBE) or partially-purified preparations reconstitutes the metastatic phenotype to that of the unextracted controls. The reconstitutive activity in partially purified preparations is quantitatively assessed as the mg amount of protein necessary to reduce the incidence of experimentally-induced pulmonary metastases by 50%. The reconstituting activity from B16-F1 displays of pI of 546 to 5.8, a mol. wt of 15 to 30 kDa, and a specific reconstituting activity of 10-4 U/mg. The mechanism of reconstitution is also under investigation. We are currently testing the hypothesis that extraction removes cell surface-associated protease inhibitors, resulting in a facilitated egress of pulmonary emboli into the surrounding tissues. Immunization of syngeneic hosts with CBE from B16-F10 prior to i.v. challenge significantly increases the incidence of experimentally-induced lung metastases, and this effect can be adoptively transferred using spleen cells. Both adherent and nonadherent splenic populations are necessary to transfer suppression. The final objective of this proposal is to delineate the cellular immune response to suppressogens in CBE, and to characterize the molecules and immune cells participating. Thus, butanol extraction of metastatic cells allows investigation of possible mechanisms involved in hematogenous metastases, the role of tumor cell surface structures in metastasis, and the effect of antigenic stimulation on the eventual development of metastases.

质膜被认为在成功的过程中发挥着重要作用 血行转移的建立。 膜上含有 1) 酶 破坏基底膜屏障所需； 2) 受体 同型和异型细胞关联； 3) 指导的抗原 特异性和非特异性免疫反应。 本研究的目的是 分离和表征影响细胞表面的部分 转移表型，使用丁醇提取的组合， 制备型等电聚焦、高性能凝胶渗透、离子 交换和疏水相互作用层析以及凝集素亲和力 色谱法。 申请人已采用非溶细胞提取法 丁醇会增加实验性肺损伤的发生率 转移。 提取增加了亚系的转移潜力 源自化学诱导的低或高转移表型 纤维肉瘤，MCA-F，或自发性黑色素瘤，B16。 短暂孵化 在粗丁醇提取物 (CBE) 或部分纯化中提取的细胞 制剂将转移表型重建为 未提取的控件。 部分纯化的重构活性 制剂定量评估为蛋白质的毫克量 减少实验性肺损伤的发生率是必要的 转移50%。 B16-F1 的重构活性显示 pI 546至5.8，摩尔。 wt 为 15 至 30 kDa，以及特定的重组 活性为10-4 U/mg。 重组机制也在制定中 调查。 我们目前正在测试提取的假设 去除细胞表面相关的蛋白酶抑制剂，从而产生 促进肺栓塞进入周围组织。 在静脉注射之前用来自 B16-F10 的 CBE 对同基因宿主进行免疫。 挑战显着增加了实验诱发的发生率 肺转移，这种效应可以通过脾脏过继转移 细胞。 贴壁和非贴壁脾群都是必需的 转移抑制。 该提案的最终目标是划定 CBE 中对抑制原的细胞免疫反应，并表征 参与的分子和免疫细胞。 因此，丁醇萃取 转移细胞可以研究参与的可能机制 血行转移，肿瘤细胞表面结构的作用 转移以及抗原刺激对最终转移的影响 转移的发展。