PATHOGENESIS OF IMMUNODEFICIENCIES IN MAN

人类免疫缺陷的发病机制

• 批准号: 3139246
• 负责人: LAWRENCE K JUNG
• 金额: $ 18.16万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-06-01 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3139246
• 关键词: CD antigens; CD3 molecule; T lymphocyte; biological signal transduction; human tissue; immunodeficiency; leukocyte activation /transformation; tissue /cell culture

DESCRIPTION (Adapted from Applicant's Abstract): CD7, is the earliest T-cell antigen to appear during human T-cell ontogeny. Although the precise biological function of CD7 is unknown, there is evidence to suggest that CD7 is involved in T-cell ontogeny and T-cell activation. Thus, CD7 may play an important role during the process of T-cell development and function. In the present proposal, this possibility will be studied by examining the mechanisms with which CD7 mediates T-cell proliferation; furthermore, an animal model will be developed so that the function of CD7 can be analyzed in vivo. The specific aims are (1) to identify and study the molecules which interact with CD7 during T-cell activation, (2) to study the mechanisms with which CD7 functions, (3) to study the genomic organization of human and mouse CD7 and (4) to study the role of CD7 molecule in mouse T-cell function and ontogeny. To accomplished these goals, molecules which interact with CD7 are identified by co-immunoprecipitation and by chemical crosslinking experiments. Recombinant CD7 protein, produced in the baculovirus expression system, will be used to isolate these molecules. The functional domains of CD7 involved in these interactions will be characterized by site-directed mutagenesis. The interacting molecules will be affinity-purified for functional and molecular characterization. The signal transduction pathways utilized by CD7 will be identified with protein kinase inhibitors; the consequence of CD7-mediated T-cell activation will be determined by flow cytometric measurements of activation molecules and by analysis of gene transcription of lymphokines and proto-oncogenes. A restriction map of the human genomic CD7 will be determined and its nucleotide sequence determined. The organization and sequence of the mouse CD7 gene will be similarly analyzed. Monoclonal (mAb) and polyclonal antibodies to murine CD7 will be produced in hamsters using synthetic CD7 peptides as immunogens. The mAb's will be used to study the distribution of CD7 antigen during T-cell development and to the expression of CD7 during embryogenesis or during bone marrow reconstitution of SCID mouse. Finally, the disruption of coding region of the CD7 gene in transgenic animals will provide a powerful model to study the role CD7 in T-cell ontogeny and T-cell function.

描述（改编自申请人的摘要）：CD 7，是最早的 T细胞抗原在人类T细胞个体发育过程中出现。 虽然 CD 7的确切生物学功能尚不清楚，但有证据表明， 表明CD 7参与T细胞个体发育和T细胞活化。 因此，CD 7可能在T细胞分化过程中发挥重要作用。 发展和功能。 在本提案中，这种可能性将 通过检查CD 7介导T细胞介导的机制来研究 增殖;此外，将开发动物模型， 可以在体内分析CD 7的功能。 具体目标是：（1） 识别和研究与CD 7相互作用的分子， 活化，（2）研究CD 7功能的机制，（3） 研究人类和小鼠CD 7的基因组结构;（4）研究 CD 7分子在小鼠T细胞功能和个体发育中的作用。 到 在完成这些目标后，鉴定了与CD 7相互作用的分子。 通过免疫共沉淀和化学交联实验。 在杆状病毒表达系统中产生的重组CD 7蛋白， 将被用来分离这些分子。 CD 7的功能结构域 参与这些相互作用的特征将是位点定向的 诱变 相互作用的分子将被亲和纯化， 功能和分子表征。 的信号转导 CD 7所利用的途径将通过蛋白激酶 抑制剂; CD 7介导的T细胞活化的结果将是 通过活化分子的流式细胞术测量和通过 淋巴因子和原癌基因的基因转录分析。 一 将确定人基因组CD 7的限制性图谱， 核苷酸序列测定。 会议的组织和顺序 将类似地分析小鼠CD 7基因。 单克隆抗体（mAb）和 将在仓鼠中使用以下方法产生鼠CD 7的多克隆抗体： 合成的CD 7肽作为免疫原。 mAb将用于研究 CD 7抗原在T细胞发育过程中的分布， 胚胎发生或骨髓中CD 7的表达 重建的SCID小鼠。 最后，破坏的编码区， 转基因动物中的CD 7基因将提供一个强有力的研究模型 CD 7在T细胞个体发育和T细胞功能中的作用。