FUNCTION AND STRUCTURE OF NOVEL HUMAN BONE MARROW CAMS

这项提案的长期目标是调查 新的骨髓基质细胞黏附分子(BM-CaM)在正常人和正常人中的表达 白血病的造血功能。二维细胞印迹技术LED的研制 对大约22个潜在的BM凸轮进行鉴定，这些摄像头显示 造血细胞黏附，更多的未成熟细胞结合到 更多种类的凸轮。其中，BM CAM 2支持单核细胞的黏附 祖细胞，包括用TNFa诱导的KG1a细胞，而BM CAM6 仅在进一步与TNFa诱导的KG1a细胞结合后才与其结合 用抗CD34或抗CD49d的CD34或CD49d进行交联刺激 MoAbs。具体目的是：1)确定粘接功能和基质 BM CAM 2和6.2的表达。2)建立BM CAM 2和 6通过测定它们的cDNAs序列。3)检验假设 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 BM-CaM-2和BM-6作为分子调控成分的基础 一个由黏附蛋白及其受体组成的互动网络。这个 研究设计和使用的方法是BM凸轮的提纯2 和6由S改编成制备级双向凝胶电泳法 技术；通过标准方法产生各自的抗体； 按既定程序克隆和测序；以及测试 BM CaM 2和6在重组BM CaM单核细胞生成中的作用。 进入最终单核细胞谱系的终点将是 巨噬细胞集落刺激因子受体(c-FMS原核表达产物)表达的检测 癌基因)，使祖细胞对M-CSF和 随后导致其他适当的细胞因子导致 成熟的单核/巨噬细胞。一种预测是， 祖细胞受体通过与BM CaM 2和6结合，导致向下- 将验证对已知PC摄像头的监管，提供可能的 成熟细胞从骨髓中释放的机制。健康- 该项目的相关性在于它提供了对基本知识的理解 正常人和白血病生长发育相关机制的研究进展 血细胞。在这个项目中产生的抗体将是 可能有助于治疗和预防白血病的传播和 淋巴瘤通过将肿瘤细胞从其转移部位转移出去； 并动员正常祖细胞进入外周血 在移植中收获和使用。

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.