REGULATION OF DENDRITIC CELL-LYMPHOCYTE INTERACTIONS

树突状细胞-淋巴细胞相互作用的调节

• 批准号: 2517099
• 负责人: CLARE J TWIST
• 金额: $ 8.93万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-01 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2517099
• 关键词: Hodgkin's disease; T lymphocyte; autoimmune disorder; blood /lymphatic neoplasm; cell cell interaction; chimeric proteins; dendritic cells; flow cytometry; gene expression; genetic mapping; glycoproteins; human genetic material tag; human tissue; in situ hybridization; laboratory mouse; laboratory rat; leukocyte activation /transformation; molecular cloning; monoclonal antibody; protein structure function; tissue /cell culture

The identification and characterization of cell surface molecules in the hematopoietic system provides insight into patterns of cellular communication and the role of these molecules in the regulation of the immune response. This project will focus on the study of HB15, a cell surface glycoprotein identified by cDNA cloning and monoclonal antibody (mAb) production and characterization. HB15 demonstrates a unique pattern of expression, being found primarily in lymphoid tissue, and restricted to cells critical in antigen (Ag)-presentation. Strong expression of HB15 has been identified on dendritic cells, Langerhans cells and Reed- Sternberg cells. Dendritic cells are potent Ag-presenting cells that are believed to be important in the activation of both cytotoxic and helper T cells. Structurally, HB15 has a single immunoglobulin (Ig)-like domain, suggesting that HB15 may be important in cellular communication. The goals of this study will be as follows: (i) To characterize the structure and function of the human and murine HB 15 gene products. The initial work will involve the molecular cloning of the murine cDNA for HB 15, and determination of the human and mouse gene structure. Identification of homologous regions should predict functional importance of the molecule and provide insight into IG gene conservation through evolution. (2) To characterize the expression pattern of this molecule in hematopoietic and histiocytic malignancies, and in particular, in Hodgkins Disease. Improved methods of dendritic cell isolation and purification will be developed using the HB 15 mAb and should provide a means for classification and characterization of the family of dendritic cells. Indirect immunofluorescence and flow cytometry analysis utilizing HB 15 mAb will be used to investigate the contribution of dendritic cells to lymphocyte activation in autoimmune disorders. (3) To characterize the function of the cells that express HB15, the role of those cells in immune dysfunction, and how HB15 is involved in the function of those cells. Dendritic cell function will be measured through assays of accessory cell function in the mixed leukocyte reaction, and the ability of HB15 mAb to abrogate this function will be tested. The ability of HB15 to provide a co-stimulatory signal for T cell proliferation will be investigated in in vitro assays. Construction of HB15-Ig fusion proteins and adhesion assays will be utilized to identify the natural ligand for HB15. These studies will provide a comprehensive effort to understand the, role of HB 15 in normal and abnormal lymphocyte biology. They will contribute to the phenotypic and functional characterization of dendritic cells and their accessory cell family members. An understanding of this molecule and the precise function it serves in the biology of lymphocyte activation will provide the basis for developing rational tools to modulate the mechanisms of lymphocyte activation in states of immune dysregulation.

细胞表面分子的鉴定和表征 在造血系统中提供了对模式的洞察 细胞通讯以及这些分子在细胞通讯中的作用 免疫反应的调节。该项目将重点关注 HB15（一种通过 cDNA 鉴定的细胞表面糖蛋白）的研究 克隆和单克隆抗体 (mAb) 生产和 表征。 HB15展示了独特的表达模式， 主要存在于淋巴组织中，仅限于细胞 对于抗原 (Ag) 呈递至关重要。 HB15的强表达 已在树突状细胞、朗格汉斯细胞和里德细胞上被鉴定出 斯滕伯格细胞。树突状细胞是有效的抗原呈递细胞 据信这对于激活两者都很重要 细胞毒性T细胞和辅助性T细胞。从结构上来说，HB15有一个单一的 免疫球蛋白 (Ig) 样结构域，表明 HB15 可能是 在细胞通讯中很重要。本研究的目标将 如下： (i) 表征结构和功能 人类和鼠类 HB 15 基因产物。初步工作将 涉及 HB 15 的鼠 cDNA 的分子克隆，以及 人类和小鼠基因结构的测定。鉴别 同源区域的数量应该预测该区域的功能重要性 分子并通过以下方式提供对 IG 基因保护的见解 进化。 (2) 表征该表达模式 造血系统和组织细胞恶性肿瘤中的分子，以及 特别是霍奇金病。树突状细胞的改进方法 将使用 HB 15 mAb 进行分离和纯化 并应提供分类和表征的方法 树突状细胞家族的成员。间接免疫荧光和 利用 HB 15 mAb 的流式细胞术分析将用于 研究树突状细胞对淋巴细胞的贡献 自身免疫性疾病的激活。 (3) 表征 表达 HB15 的细胞的功能，这些细胞在 免疫功能障碍，以及 HB15 如何参与免疫功能 那些细胞。树突状细胞功能将通过测量 混合白细胞反应中辅助细胞功能的测定， HB15 mAb 消除该功能的能力将是 已测试。 HB15 提供共刺激信号的能力 T 细胞增殖将在体外测定中进行研究。 HB15-Ig 融合蛋白的构建和粘附测定将在 用于鉴定 HB15 的天然配体。这些研究 将提供全面的努力来理解 HB 的作用 15 正常和异常淋巴细胞生物学。他们将做出贡献 树突的表型和功能表征 细胞及其辅助细胞家族成员。的理解 该分子及其在生物学中的精确功能 淋巴细胞的活化将为发育提供基础 调节淋巴细胞活化机制的合理工具 处于免疫失调状态。