IN VITRO INDUCTION OF SUPPRESSOR T CELLS

本研究提案的目的是剖析 体外产生抑制性T细胞（Ts）。 由于抑制细胞具有 与免疫系统稳态的正常维持有关， 更好地理解生成这些所需的复杂要求 重要的调节细胞群将有望使我们能够更好地 理解并纠正自身免疫性疾病、免疫缺陷 疾病和癌症。 为此，我们开发了一种体外系统， 用于产生二级（Ts 2）和效应子抑制细胞（Ts 3） 对4-羟基-3-硝基苯基乙酰半抗原（NP）具有特异性。 环磷酰胺治疗的应答者人群，其在功能上 耗尽Ts 2和Ts 3细胞，在体外用T 依赖性抗原NP-马红细胞（NP-HRBC）培养5天。 上 培养第4天，分别传代培养实验Ts 2和 将Ts 3群体加入应答培养物中。 一天后， 测定抗NP特异性空斑形成细胞（PFC）应答。 这 方法将使我们能够直接识别和充分表征两者 前体和成熟Ts群体，辅助细胞的作用，可溶性 介质，以及控制分化的遗传限制 和Ts活性的表达。 表型分析成熟和 前体Ts群体与一组单克隆试剂可以让我们 鉴定在不同Ts亚群上差异表达的标记物 并确定Ts何时发展其独特型特异性。 通过 分析胎儿肝脏和胸腺细胞群的能力， 产生Ts 2或Ts 3活动，我们将能够确定何时 抑制细胞变得具有免疫活性。 我们还将研究 辅助细胞参与Ts的产生。 我们将进行实验， 确定巨噬细胞与T或B细胞之间的MHC同源性是否 抑制细胞诱导和巨噬细胞因子是否需要 影响诱导过程。 此外，B细胞在Ts中的作用 归纳将被审查和一般问题的重要性，B 细胞受体在T细胞发育中的作用。 最后 更好地理解抑制细胞诱导的要求可以 使我们能够开发出一种方法， 抑制性T细胞系。

It is the aim of this research proposal to dissect the requirements for the in vitro generation of suppressor T cells (Ts). As suppressor cells have been implicated in the normal maintenance of immune system homeostasis, a better understanding of the complex requirements needed to generate these important regulatory cell populations will hopefully enable us to better comprehend and rectify Ts defects in autoimmune diseases, immunodeficiency disorders, and cancer. To this end we have developed an in vitro system for the generation of 2nd order (Ts2) and effector suppressor cells (Ts3) specific for the 4-hydroxy-3-nitrophenyl acetyl hapten (NP). Cyclophosphamide treated responder populations, which are functionally depleted of both Ts2 and Ts3 cells, are challenged in vitro with the T dependent antigen NP-horse red blood cells (NP-HRBC) for 5 days. On the 4th day of culture, separate subcultures containing experimental Ts2 and Ts3 populations are added to responder cultures. One day later, the direct anti-NP specific plaque forming cell (PFC) responses are determined. This methodology will allow us to directly identify and fully characterize both precursor and mature Ts populations, the role of accessory cells, soluble mediators, and the genetic restrictions which control the differentiation and expression of Ts activity. Phenotypic analysis of both mature and precursor Ts populations with a panel of monoclonal reagents may allow us to identify markers that are differentially expressed on various Ts subsets and to determine when Ts develop their idiotypic specificities. By analyzing the ability of fetal liver and thymic cell populations to generate Ts2 or Ts3 activity, we will be in a position to determine when suppressor cells become immunocompetent. We will also examine the role of accessory cells in the generation of Ts. We will perform experiments to determine whether MHC homology between the macrophages and T or B cells is required for suppressor cell induction and whether factors from macrophages influence the induction process. In addition the role of B cells in Ts induction would be examined and the general question of the importance of B cell receptors in the development of T cells can be addressed. Finally a better understanding of the requirements for suppressor cell induction may enable us to develop a method for maintaining long-term, untransformed suppressor T cell lines in culture.