MECHANISMS OF IL7-INDUCED PRE-T CELL GENE EXPRESSION

IL7诱导的前T细胞基因表达机制

• 批准号: 3455875
• 负责人: PIERETTE M APPASAMY
• 金额: $ 9.73万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 1997-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3455875
• 关键词: T cell receptor; T lymphocyte; autoradiography; cell differentiation; cell growth regulation; cell population study; embryo /fetus; gene expression; gene rearrangement; growth factor; helper T lymphocyte; hematopoietic growth factor; immunocytochemistry; in situ hybridization; laboratory mouse; leukopoiesis; liver; nucleic acid sequence; organ culture; polymerase chain reaction; southern blotting; suppressor T lymphocyte; surface antigens; thymus

The mechanisms controlling the initial aspects of thymocyte development (including rearrangement and expression of TCR genes), when pre-T cells first enter the thymus from the bone marrow or fetal liver (FL), are unknown. The broad, long-term objective of these studies is to define the mechanisms by which pre-T cell development is regulated. We found that interleukin-7 (IL7), a pre-B cell and thymocyte growth factor, stimulates growth of FL cells, expression of TCR genes, and expression of T cell- related antigens (Thy1 and PgP1) in the murine FL in vitro. Stem cell factor (SCF), a growth factor for myeloid and erythroid progenitor cells also stimulates FL cell growth and expression of cell surface TCR antigens. The specific aims of this proposal are to 1) further examine the ability of IL7 and SCF to induce: expression and rearrangement of TCR genes, intracellular and cell surface expression of T cell antigens, and cell proliferation and to determine the relationship between gamma/delta cells induced by IL7 and gamma/delta cells in the fetal thymus and peripheral organs; 2) determine whether responsiveness to IL7 and SCF is mediated directly by these factors; 3) determine whether IL7 plays a role in development of T cells in utero. These studies will establish a system for further investigation of the molecular mechanisms which regulate pre-T cell development. The experiments described herein will utilize: assays of thymic repopulation to test if IL7 and SCF affect pre-T cell activity; in situ hybridization, immunohistochemistry, and autoradiography to determine if the cells which proliferate in response to IL7 or SCF are the same cells which express T cell genes and antigens; reverse transcription and polymerase chain reaction followed by sequencing, as well s Southern blot analysis to determine which gamma and delta variable regions are expressed in response to IL7 or SCF and whether these genes are rearranged. The importance of IL7 in pre-T cell development in the fetus will be evaluated by assessing the previously mentioned endpoints following in utero injection of anti-IL7 antibodies or fetal thymus organ culture with anti- IL7 antibodies.

控制胸腺细胞发育初始方面的机制 （包括TCR基因的重排和表达），当Pre-T细胞 首先从骨髓或胎儿肝（FL）进入胸腺，是 未知。 这些研究的广泛，长期目标是定义 通过调节前T细胞发育的机制。 我们发现 白介素-7（IL7），一种前B细胞和胸腺细胞生长因子，刺激 FL细胞的生长，TCR基因的表达和T细胞的表达 体外鼠FL中的相关抗原（Thy1和PGP1）。 干细胞 因子（SCF），髓样和红细胞祖细胞的生长因子 还刺激细胞表面TCR抗原的FL细胞生长和表达。 该提案的具体目的是1）进一步研究 IL7和SCF诱导：TCR基因的表达和重排， T细胞抗原和细胞的细胞内和细胞表面表达 增殖并确定γ/三角洲细胞之间的关系 由IL7和γ/Delta细胞诱导的胎儿胸腺和周围 器官； 2）确定对IL7和SCF的响应是否介导 直接通过这些因素； 3）确定IL7是否在 子宫内T细胞的发展。 这些研究将建立一个系统 进一步研究调节前T细胞的分子机制 发展。 本文所述的实验将使用：胸腺测定 重生以测试IL7和SCF是否影响前T细胞活性；原位 杂交，免疫组织化学和放射自显影 响应于IL7或SCF的细胞是相同的细胞 表达T细胞基因和抗原；逆转录和 聚合酶链反应，然后进行测序，以及S Southern印迹 分析以确定表达哪些伽马和三角洲变量区域 响应IL7或SCF以及这些基因是否被重新排列。 这 将评估IL7在胎儿中的T PERT细胞发育中的重要性 通过评估前面提到的端点以下 注射抗IL7抗体或胎儿胸腺器官培养 IL7抗体。