INTERACTION OF T CELLS IN IMMUNITY AND TOLERANCE

该项目将继续我们对可溶性分子的研究， 在对2，4-二氯苯氧乙酸的接触超敏反应中调节T细胞免疫的T细胞 二硝基氟苯（DNFB）。 我们已经建立了一个大型小组， 克隆的抑制性T细胞（Ts）杂交瘤， I类MHC限制性抑制分子。 这些Ts杂交瘤 重排的TcR α和β链基因，并表达80-90 kd的α/β 异二聚体TcR。 抑制分子是一个100-11- kd的异源二聚体 分子，由非抗原结合（NAgB）链组成， v β和c β决定簇和抗原结合（AgB）链， 表达Calpha决定簇。 Nagb链决定MHC限制， AgB链决定抗原特异性。 通过这些参数， 抑制分子是TcR的可溶性类似物。 这项建议会 提供有关生物化学特性的更明确的信息 的抑制分子，其作用机制及其关系， TcR。 机制将通过确定的影响， 抑制分子对DNFB产生淋巴因子的各种参数的影响- 体外免疫T细胞。 还将进行研究，以确定 抑制分子来源于糖基-磷脂酰肌醇（GPI）- TcR的连接形式。 Ts杂交瘤或表达工程化的 将用磷脂酶C处理GPI连接的TcR形式， 测试上清液的抑制剂活性。 基因编码 受体和抑制分子将被克隆和测序， 转染入一组TcR α或β链缺失变体， 恢复受体表达和抑制分子产生。 最后， 抑制分子将从高滴度生物反应器中分离 用于生物化学表征、测序和晶体学分析的上清液 分析. 这些研究的结果将加深我们对如何 T细胞的功能，它们如何调节T细胞介导的免疫反应， TcR如何与抗原/MHC复合物相互作用。 另外这些 结果可能提供重要的信息有用的发展 可用于抗原特异性调节的治疗性分子 T细胞介导的自身免疫性疾病

This project will continue our studies on the soluble molecules produced by T cells which regulate T cell immunity in contact hypersensitivity to 2,4- dinitrofluorobenzene (DNFB) in mice. We have established a large panel of cloned suppressor T cell (Ts) hybridomas which produce a DNP-specific, class I MHC-restricted suppressor molecule. These Ts hybridomas have rearranged TcR alpha and beta chain genes and express a 80-90 kd alpha/beta heterodimer TcR. The suppressor molecule is a 100-11- kd heterodimer molecule and consists of a nonantigen-binding (NAgB) chain which expresses vbeta and cbeta determinants and an antigen-binding (AgB) chain which expresses Calpha determinants. The Nagb chain dictates MHC restriction and the AgB chain dictates antigen specificity. By these parameters the suppressor molecule is a soluble analogue of the TcR. This proposal will provide more definitive information concerning the biochemical properties of the suppressor molecule, its mechanism of action and its relationship to the TcR. Mechanisms will be studied by determining the effect of the suppressor molecule on various parameters of lymphokine production by DNFB- immune T cells in vitro. Studies will also be done to determine if the suppressor molecule is derived from a glycosyl-phosphatidylinositol (GPI)- linked form of the TcR. Ts hybridomas or cells expressing an engineered form of a GPI-linked TcR will be treated with phospholipase C and the supernatants tested for suppressor activity. Genes which encode the receptor and suppressor molecule will be cloned and sequenced and transfected into a panel of TcR alpha or beta chain deleted variants to restore receptor expression and suppressor molecule production. Finally, the suppressor molecule will be isolated from high titered bioreactor supernatants for biochemical characterization, sequence and crystalographic analysis. Results from these studies will enhance our understanding of how Ts cells function, how they regulate T cell-mediated immune responses and how the TcR interacts with antigen/MHC complexes. In addition, these results may provide important information useful for the development of therapeutic molecules which may be used for the antigen-specific regulation of debilitating T cell-mediated autoimmune disease