CHARACTERIZATION OF LYMPHOBLAST PEPTIDE RECEPTORS

淋巴细胞肽受体的表征

• 批准号: 3182952
• 负责人: STEPHEN D SMITH
• 金额: $ 16.07万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-30 至 1990-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3182952
• 关键词: aminoacid transport; antineoplastics; cell differentiation; cell growth regulation; chromosome disorders; cytogenetics; electrofocusing; flow cytometry; gel electrophoresis; gel filtration chromatography; glucose transport; hematopoietic stem cells; immunofluorescence technique; immunoprecipitation; insulin; insulinlike factor; lymphoblast; neoplasm /cancer therapy; neoplastic growth; peptide hormone; phosphorylation; radioimmunoassay; radiotracer; tissue /cell culture

In the past, we developed an assay which supports the growth of malignant colonies from children with acute lymphoblastic leukemia and non-Hodgkin's lymphoma. Our methodology differs from in vitro culture techniques reported by others in that (albeit sporadically) long term cell growth and cell line establishment from passage of a single colony could be accomplished. Cell line establishment, however, is important because cell lines provide an unlimited, stable source of cells for in vitro studies. Cell lines established in our laboratory have been shown to have the same immunophenotype, biochemical enzyme profile, immunogenotype and karyotype as the patient's tumor. We have recently determined that different subtypes of malignant lymphoblasts possess unique profiles for receptors to peptide hormones. While each of 9 immature T-lymphoblast cell lines possess IGF-I receptors and lack insulin receptors, 19 of 20 immature B-lymphoblast cell lines possess insulin receptors and lack IGF-I receptors. We have shown that T-lymphoblasts proliferate in response to nanogram concentrations of IGF-I but not insulin in serum-free media. Moreover, supplemental IGF-I has proven to be a key growth factor necessary in the development of a reproducible, standardized method for the establishment of T-lymphoblast cell lines in vitro. Based on these data, we hypothesize that peptide hormones such as insulin and IGF-I may be critical growth factors necessary for the proliferation of malignant hematopoietic cells and that peptide receptor function blockage may severely limit cellular proliferation. The objective of this proposal is to study the biologic function of specific peptides (I, IGF-I, IGF-II, CH) on malignant hematopoietic cells which possess (or lack) specific peptide receptors, and to determine the specificity, kinetics, structure and function on specific peptide receptors by binding studies, chemical cross-linking on SDS-PAGE, receptor phosphorylation and by polyclonal and monoclonal antireceptor antibodies. These studies will augment our understanding of peptide/peptide receptor function in malignant cells, will help establish standard methodologies for cell line establishment, and may provide the foundation for antipeptide receptor therapy for hematopoietic malignancies in the future.

在过去，我们开发了一种支持生长的测定方法， 急性淋巴细胞白血病患儿的恶性集落 白血病和非霍奇金淋巴瘤 我们的方法不同 来自其他人报道的体外培养技术（尽管 零星）长期细胞生长和细胞系建立 从一个单一的殖民地的通道可以完成。 细胞系 然而，建立是重要的，因为细胞系提供了 为体外研究提供无限、稳定的细胞来源。 细胞系 在我们实验室建立的实验室已经被证明具有相同的 免疫表型、生化酶谱、免疫基因型 和染色体组型一样 我们最近发现，不同亚型的恶性肿瘤 淋巴母细胞具有独特的肽受体谱 荷尔蒙 9种未成熟T淋巴母细胞系 有IGF-I受体和缺乏胰岛素受体，19 20 未成熟B淋巴母细胞系具有胰岛素受体， 缺乏IGF-I受体。 我们已经证明T淋巴母细胞 对纳克浓度的IGF-I有反应， 而不是无血清培养基中的胰岛素。 此外，补充IGF-I 已被证明是一个关键的增长因素， 开发可重复的标准化方法， 体外T淋巴母细胞系的建立。 基于这些 数据，我们假设肽激素如胰岛素和 IGF-I可能是关键的生长因子， 恶性造血细胞增殖与肽 受体功能阻断可能会严重限制细胞 增殖 这项建议的目的是研究 特异性肽（I，IGF-I，IGF-II，CH）对 具有（或缺乏）特异性的恶性造血细胞 肽受体，并确定特异性，动力学， 结构和功能对特异性肽受体的影响 研究，SDS-PAGE化学交联，受体 磷酸化和通过多克隆和单克隆抗受体 抗体的 这些研究将增加我们对 肽/肽受体功能的恶性细胞，将有助于 建立用于细胞系建立的标准方法，以及 可能为抗肽受体治疗提供基础， 造血系统恶性肿瘤