PATHOGENESIS OF IMMUNODEFICIENCIES IN MAN

描述(改编自申请人摘要)：CD7，是最早的 T细胞抗原出现在人类T细胞个体发育过程中。尽管 CD7的确切生物学功能尚不清楚，有证据表明 提示CD7参与T细胞个体发育和T细胞活化。 因此，CD7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，这一可能性将 通过检测CD7介导T细胞的机制来研究 此外，还将开发一种动物模型，以便 CD7的功能可以在体内进行分析。具体目标是(1) 鉴定和研究T细胞中与CD7相互作用的分子 激活，(2)研究CD7的作用机制，(3) 研究人和小鼠CD7的基因组组织和(4)研究 CD7分子在小鼠T细胞功能和个体发育中的作用至 完成这些目标后，识别出与CD7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CD7蛋白， 将被用来分离这些分子。CD7的功能结构域 参与这些交互的特征将是站点定向 诱变。相互作用的分子将被亲和纯化以 功能和分子表征。信号转导 CD7利用的通路将被蛋白激酶识别 抑制剂；CD7介导的T细胞激活的后果将是 由激活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。一个 将确定人类基因组CD7的限制性内切酶图谱，并将其 核苷酸序列已确定。会议的组织和顺序 小鼠的CD7基因也将进行类似的分析。单抗(MAb)和 将在仓鼠体内生产针对小鼠CD7的多克隆抗体 人工合成的CD7多肽作为免疫原。单抗将被用来研究 CD7抗原在T细胞发育过程中的分布 CD7在胚胎发育和骨髓中的表达 SCID小鼠的重建。最后，基因编码区的破坏 转基因动物中的CD7基因将为研究提供一个强有力的模型 CD7在T细胞个体发育和T细胞功能中的作用

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.