FUNCTIONAL ANALYSIS OF HUMAN HELPER T CELL CLONES

在过去的三年里，我们实验室的注意力集中在 克隆抗原特异性的分离和长期体外生长 作为白细胞介素-2（IL-2）依赖性T细胞系增殖的人T细胞 （TCL）。 我们对分析T细胞特异性 对于半抗原（三硝基苯基（TNP））修饰的自体细胞，作为 人T细胞对病毒和肿瘤抗原免疫。 过程中 通过这些研究，我们已经成功地建立了长期培养克隆 辅助TCL特异于多种抗原，包括：半抗原， 可溶性抗原和同种异体抗原。 此外， 这些克隆的TCL已经确定了几种新的免疫调节途径， 其控制体液和细胞介导的免疫反应。 主要 本提案的主旨将是继续界定 特异性、遗传限制和调控的作用机制 人T细胞克隆。 具体而言，我们的目标是：1）改善文化 克隆的人T细胞的长期IL-2依赖性生长的方法 细胞，特别强调优化条件， 抑制细胞的生长; 2）分析可溶性因子并直接 抗原特异性辅助性T细胞克隆 与各种B细胞亚群、巨噬细胞和其它T细胞协作; 这将包括抗原特异性辅助T细胞的研究， 靶向扩增体液或细胞溶解性T细胞（CTL）应答;和 3)利用克隆的自体抗原特异性TCL作为免疫原 体外产生抗自身独特型T细胞克隆，其下调 以抗原特异性方式产生免疫应答。 使用的测定系统、生成的克隆TCL和信息 在正常的调节相互作用的研究中获得的， 自然地，更精确地分析人类疾病状态， 免疫调节紊乱;特别是自身免疫状态， 免疫缺陷综合征，包括艾滋病。

Over the past three years, the attention of our laboratory has been focused on the isolation and long term in vitro growth of cloned antigen specific human T cells propagated as interleukin-2 (IL-2) dependent T cell lines (TCL). We have been particularly interested in analyzing T cells specific for hapten (trinitrophenyl (TNP)) modified autologous cells, as a model of human T cell immunity to virus and tumor antigens. During the course of these studies, we have successfully established in long term culture cloned helper TCL specific for a wide variety of antigens, including: hapten, soluble antigens, and alloantigens. Moreover, the functional analysis of these cloned TCL has identified several novel immunoregulatory pathways which govern both humoral and cell mediated immune reactions. The major thrust of the present proposal will be to continue to define the specificity, genetic restrictions, and mechanisms of action of regulatory human T cell clones. Specifically, our goals are to: 1) improve culture methodologies for the long term IL-2 dependent growth of cloned human T cells, with particular emphasis on optimizing conditions for the consistent growth of suppressor cells; 2) analyze the soluble factor(s) and direct cell-cell interactions by which antigen specific helper T cell clones collaborate with various B cell subsets, macrophages, and other T cells; this will include the study of antigen specific helper T cells specifically targeted to amplify either humoral or cytolytic T cell (CTL) responses; and 3) utilize cloned autologous antigen specific TCL as immunogens for the in vitro generation of anti-auto idiotypic T cell clones which down regulate immune responses in an antigen specific manner. The assay systems utilized, the cloned TCL generated, and the information gained in the study of normal regulatory interactions should extend naturally to a more precise analysis of human disease states characterized by disordered immunoregulation; specifically autoimmune states and the spectrum of immunodeficiency syndromes, including AIDS.