T CELL COLONY FORMATION IN NORMALS AND AIDS

T细胞集落测定是一种有价值的评估因子的技术 控制着人类正常的淋巴生成。在此过程中，单个核细胞 细胞或分离的T淋巴细胞在半固体培养液中培养并生长 是由一种非特异的有丝分裂原刺激的，通常是PHA。此前，我们有 研究表明，PHA刺激的分离T细胞的克隆性增殖依赖于 在可溶的引发因素存在的情况下。IL-1和IL-2都会 从属于这个功能。此外，肿瘤产生的启动因子 B淋巴细胞和单核细胞系的细胞系都会促进集落形成 成长。值得注意的是，这些肿瘤的因子似乎不同于 经典的Interleukins。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。这个 具体目标包括：(1)生化特性和纯化 由Raji细胞(A B淋巴瘤系)阐述的可溶性因子，一种有效的 菌落反应的启动子和从其他细胞获得的上清液 系列(2)用单抗分离的T细胞亚群的测定 抗体技术，能够形成克隆(3)评估 单个核细胞特定亚群的潜在调节作用 以及来自这些细胞的可溶性产物对T细胞生长的影响，(4) 免疫抑制药物对克隆性细胞的活性检测 反应(5)测量所含细胞的功能活动 通过在含有IL-2的培养液中扩增它们并确定 它们在商陆诱导的免疫球蛋白合成实验中的活性 (6)佛波酯有丝分裂活性的比较 (TPA)和琼脂糖与PHA结合的T3。这些化合物似乎激活了 与PHA表达的机制不同。第二 这项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 评估ALL临床患者中出现的抑制的菌落反应 HTLV-III感染的阶段。初步结果表明，有几个 这些因素可能是导致这种综合征的生长缺陷的原因； 这些包括IL-2的产生减少，能力的缺陷 细胞表达IL-2受体及其存在的各种抑制作用 各种因素。拟议的研究将确定负责 这种生长异常。此外，旨在确定 将进行菌落分析的预后价值评估。

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.