CD7 AND LYMPHOCYTE DEVELOPMENT AND FUNCTION

CD7 和淋巴细胞的发育和功能

• 批准号: 2002697
• 负责人: FRACISCO A BONILLA
• 金额: $ 8.8万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2002697
• 关键词: B lymphocyte; CD antigens; CD8 molecule; T lymphocyte; biological signal transduction; cell differentiation; chimeric proteins; flow cytometry; gene targeting; immunocytochemistry; laboratory mouse; molecular cloning; natural killer cells; nucleic acid sequence; tissue /cell culture; yeast two hybrid system

Francisco A. Bonilla holds both M.D. and Ph.D. (Immunology) degrees. Following clinical training in Pediatrics, and a fellowship in Allergy/Immunology at Children's Hospital, Boston, he has recently been appointed to the faculty of the Division of Immunology at this institution. Dr. Bonilla now plans to continue as a Scientist Physician performing basic Immunology research, with the eventual goal of becoming an independent investigator. Dr. Bonilla's work will be carried out within the research facilities of the Division of Immunology of Children's Hospital. A large body of literature strongly suggests that the lymphocyte surface marker CD7 has an important role in the development and function of a number of leukocyte lineages. However, the ligand for CD7 and its signaling pathways remain unknown. We will use the method of targeted gene disruption to create mice lacking CD7. This will provide a system in which to clearly define the importance of CD7 in the development and function of the cells which express it CD7-deficient mice will be studied with respect to the representation of lymphocyte subpopulations in primary and secondary lymphoid tissues by flow cytometry, and immunohistochemical methods. They will be challenged in vivo with several antigens and their antibody responses assessed. They will also be subjected to a comprehensive battery of in vitro tests of T cell, B cell, and natural killer cell function. We will also study the interaction of the intracellular portion of CD7 with signaling pathways. We will create a number of hybrid constructs consisting of the external portion of the human CD8 molecule joined to the cytoplasmic domain of human CD7. The CD7 domain will be subjected to various mutations and deletions; cells transfected with these constructs will be studied with respect to several biological effects associated with CD7 ligation. In addition, we will employ the yeast two hybrid system to identify proteins associated with the intracellular domain of CD7. Interacting proteins will be cloned and sequenced. We will attempt to determine their potential roles in signaling through structural comparisons with known proteins, as well as by analyzing phosphorylation patterns following cellular stimulation.

弗朗西斯科A.博尼拉拥有两个医学博士学位。和博士免疫学学位。 经过儿科临床培训， 过敏/免疫学在儿童医院，波士顿，他最近被 被任命为免疫学系的教师， 机构。Bonilla博士现在计划继续担任科学家医师 进行基本的免疫学研究，最终目标是 成为独立调查员。博尼利亚博士的工作将是 在海洋事务司的研究设施内进行的 儿童医院免疫科。 大量文献强烈表明，淋巴细胞 表面标志物CD 7在发育中具有重要作用， 许多白细胞谱系的功能。然而，CD 7的配体 它的信号通路仍然未知。我们将使用的方法 靶向基因破坏以产生缺乏CD 7的小鼠。这将提供 一个系统，其中明确定义的重要性，CD 7在 表达CD 7缺陷型的细胞的发育和功能 将研究小鼠淋巴细胞的代表性 按血流分析的初级和次级淋巴组织中的亚群 流式细胞术和免疫组织化学方法。他们将面临挑战， 用几种抗原进行体内试验并评估其抗体应答。他们 还将接受一系列全面的体外测试， T细胞、B细胞和自然杀伤细胞功能。 我们还将研究CD 7的细胞内部分的相互作用， 信号通路。我们将创造一些混合结构 由人CD 8分子的外部部分连接到 人CD 7的胞质结构域。CD 7结构域将受到 各种突变和缺失;用这些转染的细胞 将研究构建体的几种生物学效应 与CD 7连接有关。 此外，我们还将利用酵母双杂交系统进行鉴定， 与CD 7的胞内结构域相关的蛋白质。交互 蛋白质将被克隆和测序。我们将试图确定 它们在信号传导中的潜在作用，通过与 已知的蛋白质，以及通过分析磷酸化模式 在细胞刺激之后。