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 克隆抗原特异性克隆的分离及体外长期生长的研究 人T细胞作为白介素2依赖的T细胞株的增殖 (TCL)。我们对分析特定的T细胞特别感兴趣 对于半抗原(三硝基苯基)修饰的自体细胞，作为 人类T细胞对病毒和肿瘤抗原的免疫。在…的过程中 这些研究表明，我们已经成功建立了长期克隆培养体系 辅助TCL对多种抗原具有特异性，包括：半抗原， 可溶性抗原和同种异体抗原。此外，还对其进行了功能分析。 这些克隆的TCL已经确定了几个新的免疫调节途径 它既控制体液免疫反应，也控制细胞免疫反应。少校 本提案的主旨将是继续界定 调节性基因的特异性、遗传限制和作用机制 人类T细胞克隆。具体地说，我们的目标是：1)提升文化 克隆人T细胞长期依赖IL-2生长的方法 电池，特别强调优化条件，以保持一致 抑制细胞的生长；2)分析可溶性因子(S)和直接 抗原特异性辅助性T细胞克隆的细胞-细胞相互作用 与各种B细胞亚群、巨噬细胞和其他T细胞合作； 这将包括对抗原特异性辅助T细胞的研究 目标是放大体液或细胞溶解T细胞(CTL)反应；以及 3)利用克隆的自体抗原特异性TCL作为免疫原 体外制备下调表达的抗自身独特型T细胞克隆 以抗原特异的方式进行的免疫反应。 所使用的检测系统、克隆的TCL的生成和信息 在研究正常的监管相互作用方面取得的成果应该扩大 自然能够更精确地分析人类疾病状态的特点 通过紊乱的免疫调节；特别是自身免疫状态和 免疫缺陷综合征的范围，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.