REGULATION OF DENDRITIC CELL-LYMPHOCYTE INTERACTIONS

细胞表面分子的鉴定和表征 在造血系统中的作用提供了对 细胞通讯以及这些分子在 调节免疫反应。该项目将侧重于 细胞表面糖蛋白HB 15 cDNA研究 克隆和单克隆抗体（mAb）生产， 特征化HB 15显示出独特的表达模式， 主要存在于淋巴组织中，仅限于细胞 在抗原（Ag）呈递中至关重要。HB 15的强表达 已经在树突状细胞、朗格汉斯细胞和里德细胞上被鉴定出来， 斯滕贝格细胞。树突状细胞是一种强有力的抗原提呈细胞 这被认为是重要的激活两个 细胞毒性和辅助性T细胞。从结构上讲，HB 15有一个单一的 免疫球蛋白（IG）样结构域，表明HB 15可能是 在蜂窝通信中很重要。本研究的目标将 （一）对专利权的保护; 人和鼠HB 15基因产物。初步工作将 涉及HB 15的鼠cDNA的分子克隆，和 确定人类和小鼠的基因结构。识别 同源区域的功能重要性， 分子，并提供深入了解IG基因保守， 进化(2)为了表征这种表达模式， 分子在造血和组织细胞恶性肿瘤中的作用， 特别是在霍奇金斯病中。树突状细胞的改良方法 将使用HB 15 mAb开发分离和纯化 并应提供一种分类和表征的方法 树突状细胞家族的成员。间接免疫荧光和 使用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 自身免疫性疾病中的活化。(3)表征 表达HB 15的细胞的功能，这些细胞在 免疫功能障碍，以及HB 15是如何参与的功能， 这些细胞。树突状细胞功能将通过 混合白细胞反应中辅助细胞功能的测定， 而HB 15 mAb消除该功能的能力将是 测试. HB 15提供共刺激信号的能力， 将在体外测定中研究T细胞增殖。 HB 15-IG融合蛋白的构建和粘附试验将在 用于鉴定HB 15的天然配体。这些研究 将提供一个全面的努力，了解HB的作用， 15在正常和异常淋巴细胞生物学。他们将作出贡献 树突状细胞的表型和功能特征 细胞及其附属细胞家族成员。了解 这种分子及其在生物学中的确切功能， 淋巴细胞活化将为发展 调节淋巴细胞活化机制的合理工具 处于免疫失调状态

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.