MECHANISMS OF T-LYMPHOCYTE DEVELOPMENT

总的目标是定义参与的细胞和分子事件 胸腺微环境-胸腺细胞相互作用导致T细胞正常 发育或导致T细胞异常成熟，如在 先天性和获得性T细胞免疫缺陷状态，以及T细胞 恶性肿瘤。实现这一总体目标所需的研究包括 两个主要领域--上皮细胞的表型和功能特征 人类胸腺微环境中的细胞，以及表型和 人T细胞及其前体细胞的功能特性 确定的成熟阶段。胸腺上皮成熟的研究 人类内分泌胸腺上皮的角蛋白亚类将被用作 胸腺微环境成熟的标志物。使用一种新开发的 胸腺上皮细胞与T细胞结合的测定，我们将研究和定义 功能相关的胸腺上皮表面分子与 胸腺上皮-T细胞结合、胸腺激素分泌和 分泌其他免疫调节分子。表型和 恶性胸腺上皮细胞(胸腺瘤)的分化能力 与正常和增生性肌无力相比，重症肌无力将使用 我们最近开发的人类长期体外生长技术 胸腺上皮。此外，EGF非依赖性转化的胸腺 通过引入转化基因来培育上皮细胞系 (n-ras，c-Myc)转化培养的正常胸腺上皮细胞。使用人类 恶性多能干细胞系(DP.528)，我们将开发出特异性的 定义表面或细胞质抗原的单抗探针 早期特异性(多能干细胞、淋巴干细胞、承诺T细胞 细胞前体、原胸腺细胞)T细胞成熟阶段。最后，我们 将开发细胞、分子和基因的体外检测系统 在T细胞成熟的早期阶段可能发生的事件。使用 这些检测系统，培养的胸腺上皮的影响，其他 辅助细胞，胸腺上皮上清液，合成胸腺激素， 纯化的白介素1或白介素2对细胞分化能力的影响 我们将研究未成熟、正常和恶性的T细胞。

The general goal is to define cellular and molecular events involved in thymic microenvironment-thymocyte interactions that lead to normal T cell development or lead to aberrant T cell maturation, such as is seen in congenital and acquired T cell immunodeficiency states, and in T cell malignancies. The studies required to achieve this general goal fall into two main areas--phenotypic and functional characterization of epithelial cells within the human thymic microenvironment, and phenotypic and functional characterization of human T cells and their precursors at defined stages of maturation. To study thymic epithelial maturation keratin subclasses of human endocrine thymic epithelium will be used as markers of thymic micro-environment maturation. Using a newly developed assay of thymic epithelial-T cell binding, we will study and define functionally relevant thymic epithelial surface molecules with regard to thymic epithelial-T cell binding, secretion of thymic hormones and secretion of other immunoregulatory molecules. The phenotype and differentiating capacity of malignant thymic epithelial cells (thymomas) compared to normal and hyperplastic myasthenia gravis will be studied using our recently developed techniques for long term in vitro growth of human thymic epithelium. In addition, EGF-independent transformed thymic epithelial cell lines will be developed by introducing transforming genes (N-ras, c-Myc) into cultured normal thymic epithelial cells. Using a human malignant pluripotent stem cell line (DP.528), we will develop specific monoclonal antibody probes that define surface or cytoplasmic antigens specific for early (pluripotent stem cell, lymphoid stem cell, committed T cell precursor, prothymocyte) stages of T cell maturation. Finally, we will develop in vitro assay systems for cellular, molecular and genetic events that might transpire during early stages of T cell maturation. With these assay systems, the effects of cultured thymic epithelium, other accessory cells, thymic epithelial supernatants, synthetic thymic hormones, purified Interleukin 1 or Interleukin 2 on the differentiating capacity of immature normal and malignant T cells will be studied.