T CELL COLONY FORMATION IN NORMALS AND AIDS

T细胞集落试验是一种有价值的评估因素的技术 控制正常的人类淋巴细胞生成 在这个过程中，单核细胞 将细胞或分离的T淋巴细胞接种在半固体培养基中， 是由一种非特异性的有丝分裂原刺激的，通常是PHA。 此前，我们有 表明PHA刺激的分离T细胞的克隆扩增依赖于 在可溶性起始因子的存在下。 IL-1和IL-2都将 支持这个功能。 此外，肿瘤产生的启动因子 B淋巴细胞和单核细胞谱系的细胞系将促进集落形成 增长 值得注意的是，这些肿瘤的因素似乎与 经典的白细胞介素 拟议的研究旨在进一步 确定克隆生长所需的细胞和可溶性因子。 的 具体目标包括：（1）生物化学表征和纯化 Raji细胞（一种B淋巴瘤细胞系）产生的可溶性因子， 集落应答起始物和从其它细胞获得的上清液 （2）T细胞亚群的测定，通过单克隆抗体分离， 抗体技术，能够形成集落（3）评价 单核细胞特定亚群的潜在调节作用 以及来源于这些细胞的可溶性产物对T细胞生长的影响，（4） 免疫抑制剂对克隆形成的抑制作用 （5）测量所含细胞的功能活性 通过在含有IL-2的培养基中扩增菌落并测定 它们在美洲商陆诱导的免疫球蛋白合成测定中的活性， (6)佛波醇肉豆蔻酸酯的促有丝分裂活性比较 (TPA)琼脂糖结合T3与PHA。 这些化合物似乎能激活 通过不同于PHA表达的那些机制来表达T细胞。 第二 本研究的主要方面是评估与以下相关的克隆反应： 几种疾病。 这些研究的一个主要组成部分是进一步 评价在所有临床表现的患者中观察到的抑制集落反应， HTLV-III感染的阶段。 初步结果显示， 因素可能是负责在这种综合征中看到的生长缺陷; 这些包括IL-2的产生减少， 细胞表达IL-2受体和存在各种抑制性 因素 拟议的研究将确定负责 这种生长异常 此外，旨在确定 将进行殖民地测定的预后价值。

The T cell colony assay is a valuable technique for assessing factors controlling normal human lymphopoiesis. In this procedure, mononuclear cells or isolated T lymphocytes are plated in semi-solid media and growth is stimulated by a non-specific mitogen, usually PHA. Previously, we have shown that the clonal expansion of PHA-stimulated isolated T cells depends on the presence of a soluble initiating factor. Both IL-1 and IL-2 will subserve this function. In addition, initiating factors produced by tumor cell lines of both B lymphocytic and monocytic lineages will promote colony growth. Of note, the factors from these tumors appear to be different than the classic interleukins. The proposed studies are designed to further define the cellular and soluble factor requirements for clonal growth. The specific aims include: (1) biochemical characterization and purification of the soluble factor elaborated by Raji cells (a B lymphoma line), a potent initiator of colony responses, and supernatants obtained from other cell lines (2) determinations of the T cell subsets, separated by monoclonal antibody techniques, capable of forming colonies (3) evaluations of the potential regulatory influences of specific subsets of mononuclear cells and soluble products derived from these cells on the T cell growth, (4) examinations of the activities of immunosuppressive drugs on clonal responses (5) measurements of the functional activities of cells contained in colonies by expanding them in IL-2 containing media and determining their activities in the pokeweed-induced immunoglobulin synthesis assay and (6) comparisons of the mitogenic activities of phorbol myristate acetate (TPA) and Sepharose-bound T3 with PHA. These compounds appear to activate T cells by mechanisms distinct from those expressed by PHA. The second major aspect of this study is to evaluate clonal responses associated with several disorders. A principal component of these studies is to further evaluate the suppressed colony responses seen in patients with all clinical phases of HTLV-III infections. Initial results suggest that several factors may be responsible for the growth defects seen in this syndrome; these include decreased productions of IL-2, deficits in the ability of cells to express IL-2 receptors and the presence of various inhibitory factors. The proposed studies will define the mechanisms responsible for this growth abnormality. In addition, studies aimed at determining the prognostic value of the colony assay will be performed.