FUNCTION AND STRUCTURE OF NOVEL HUMAN BONE MARROW CAMS

本提案的长期目标是研究 在正常人和正常人中发现新的骨髓基质细胞粘附分子（BM CAM）， 白血病造血二维细胞印迹技术的发展 鉴定了大约22种潜在的新型BM CAM， 造血细胞粘附，更多的未成熟细胞结合到一个 更多种类的CAM其中，BM CAM 2支持单核细胞粘附， 祖细胞，包括用TNF α诱导的KG 1a细胞，而BM CAM 6 结合TNF α诱导的KG 1a细胞后，他们已经进一步 通过使用抗-CD 34或抗-CD 49 d交联CD 34或CD 49 d刺激 MoAbs具体目的是：1）确定粘附功能和基质 BM CAM 2和6的表达。 2)确定BM CAM 2的身份， 6的cDNA序列。 3)测试的假设 具有单核细胞分化潜能的早期祖细胞与 BM CAM 2和/或BM CAM 6促进单核细胞发育，这是基于 BMCAMs 2和6作为BMCAMs的分子调节组分发挥作用的基础。 粘附蛋白及其受体的相互作用网络。的 研究设计和使用的方法是纯化BM CAMs 2 和6通过适应性水平的2-D凝胶电泳 通过标准方法产生它们各自的抗体; 通过既定程序进行cDNA克隆和测序;并测试 BM CAM 2和6在单核细胞生成中的作用。 进入确定的单核细胞谱系的终点将是 评估M-CSF受体的表达（由c-fms proto编码， 癌基因），其使得祖细胞对M-CSF应答， 随后是其他适当的细胞因子，导致 成熟单核细胞/巨噬细胞。预测说， 祖细胞受体，通过与BM CAM 2和6结合， 已知的PC CAM的监管将得到验证，提供一个可能的 从骨髓中释放成熟细胞的机制。健康- 该项目的相关性在于它提供了对基本 与正常和白血病细胞生长发育相关的机制 血细胞在这个项目中产生的抗体 潜在地可用于治疗和预防白血病的扩散， 通过从肿瘤细胞的转移部位去除肿瘤细胞来治疗淋巴瘤; 以及用于将正常祖细胞动员到外周血中， 收获并用于移植。

The lone-term objective of this proposal is to investigate the role of novel bone marrow stromal cell adhesion molecules (BM CAMs) in normal and leukemic hematopoiesis. Development of a 2-D cell blotting technique led to identification of about 22 potentially novel BM CAMs that show hematopoietic cell adhesion, with the more immature cells binding to a greater variety of CAMs. Of these, BM CAM 2 supports adhesion of monocytic progenitors, including KG1a cells induced with TNFa, whereas, BM CAM 6 binds to TNFa-induced KG1a cells only after they have been further stimulated by cross-linking of CD34 or CD49d using anti-CD34 or anti-CD49d MoAbs. The specific aims are to: 1) Confirm adhesive function and stromal expression of BM CAMs 2 and 6. 2) Establish the identity of BM CAMs 2 and 6 by determining their cDNA sequences. 3) Test the hypothesis that binding of early progenitors with monocytic differentiation potential to BM CAM 2 and/or BM CAM 6 promotes monocytic development, predicated on the basis that BM CAMs 2 and 6 function as molecular regulatory components of an interactive network of adhesive proteins and their receptors. The research design and the methods to be used are purification of BM CAMs 2 and 6 by an adaptation(s) to preparative-level 2-D gel electrophoretic techniques; generation of their respective antibodies by standard methods; cDNA cloning and sequencing by established procedures; and testing the role of BM CAMs 2 and 6 in monocytopoiesis using the recombinant BM CAMs. The endpoint for entry into the definitive monocyte lineage will be assessment of expression of the M-CSF receptor (coded for by c-fms proto- oncogene), which renders the progenitor cells responsive to M-CSF and subsequently to other appropriate cytokines leading to the development of mature monocytes/ macrophages. The prediction that the engagement of progenitor cell receptors, by binding to BM CAMs 2 and 6, results in down- regulation of the known PC CAMs will be verified, providing a possible mechanism for the release of matured cells from the BM. The health- relatedness of the project is that it provides an understanding of basic mechanisms related to the growth and development of normal and leukemic blood cells. The antibodies generated during this project would be potentially useful for treating and preventing the spread of leukemias and lymphomas by dislodging the neoplastic cells from their metastatic sites; and for mobilizing normal progenitors into the peripheral blood for harvesting and use in transplantation.