STRUCTURE AND FUNCTION OF CD6 IN THE MOUSE

小鼠 CD6 的结构和功能

• 批准号: 2376424
• 负责人: JANE R PARNES
• 金额: $ 20.35万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2376424
• 关键词: CD antigens; CD5 molecule; T lymphocyte; antibody formation; antiserum; biological signal transduction; chimeric proteins; complementary DNA; embryonic stem cell; genetically modified animals; laboratory mouse; laboratory rat; leukocyte activation /transformation; monoclonal antibody; protein structure function

Anti-CD6 monoclonal antibodies and immunotoxins have been used as immunosuppressive therapies for the treatment of renal allograft rejection and graft-versus-host disease. In particular, purging of donor marrow with anti-CD6 reagents has prevented graft-versus-host disease without significantly lowering the incidence of engraftment in allogeneic bone marrow transplantation. Thus, such reagents may be of important therapeutic value in treatment of malignancies and other disorders by allogeneic bone marrow transplantation. There is thus far only limited knowledge about the biological function of CD6 except that it plays some role in T cell activation. Furthermore, a mouse homolog of CD6 has not been identified to date. The goal of this study is to analyze the structure and function of CD6 in the mouse system. This will be done by further characterization and sequencing of isolated cDNA and genomic clones encoding this protein. Monoclonal antibodies specific for mouse CD6 will be generated using the gene gun to immunize rats followed by fusion of the spleen cells, selection and screening for appropriate antibody-secreting hybridomas. These antibodies will be used to determine the tissue and cell type distribution of mouse CD6 expression as well as the pattern and level of expression during thymocyte development. They will also be used to determine their functional effects upon mouse T cells, in particular as costimulatory reagents for T cell activation. The pathway of signal transduction through the long cytoplasmic tail of this protein will be approached by using the yeast-two-hybrid system to isolate clones encoding proteins that interact with this cytoplasmic tail and by examining effects of progressive deletions in the cytoplasmic tail on the ability of CD6 to costimulate T cell activation. The physiological importance of this molecule will be further examined by the generation of mutant mice in which the CD6 gene has been knocked out by homologous recombination. The effects of lack of CD6 expression upon T cell development and activation will be examined.

抗CD6单抗和免疫毒素已被用作 免疫抑制疗法在移植肾治疗中的应用 排斥反应和移植物抗宿主病。特别是，清除捐赠者 含有抗CD6试剂的骨髓可以预防移植物抗宿主病 而不显著降低同种异体移植的发生率 骨髓移植。因此，这样的试剂可能很重要。 中西医结合治疗恶性肿瘤等疾病的治疗价值 异基因骨髓移植。到目前为止，只有有限的 关于CD6的生物学功能的知识，除了它扮演一些 在T细胞活化中的作用。此外，CD6的小鼠同源物还没有 到目前为止已经被确认。这项研究的目的是分析 CD6在小鼠体内的结构和功能。这将通过以下方式完成 分离的cDNA和基因组的进一步鉴定和测序 编码这种蛋白质的克隆。小鼠特异性单抗 将使用基因枪产生CD6来免疫大鼠，然后 脾细胞的融合，筛选合适的 分泌抗体的杂交瘤。这些抗体将被用来确定 小鼠CD6表达的组织和细胞类型分布 胸腺细胞发育过程中的表达模式和水平。他们 也将用于确定它们对小鼠T细胞的功能影响 细胞，特别是作为T细胞激活的共刺激试剂。 细胞质长尾巴的信号转导途径 这种蛋白质将通过使用酵母双杂交系统来接近 分离编码与该细胞质尾巴相互作用的蛋白质的克隆 并通过检测细胞质尾部的渐进性缺失的影响 CD6共刺激T细胞活化的能力。这个 这种分子的生理重要性将由 CD6基因被敲除的突变小鼠的产生 同源重组。CD6表达缺失对T细胞的影响 将检查细胞的发育和激活情况。