KAPPA GENE REARRANGEMENT IN TRANSFORMED PREB CELLS

转化前细胞中的 KAPPA 基因重排

• 批准号: 2071594
• 负责人: NAOMI ROSENBERG
• 金额: $ 17.84万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-05-01 至 1997-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2071594
• 关键词: Abelson leukemia virus; B lymphocyte; cell differentiation; cell line; differentiation antigens; gene expression; gene rearrangement; immunoglobulin genes; laboratory mouse; tissue /cell culture

Immunoglobulin gene rearrangement is fundamental to the development of lymphocytes. Understanding the generation of these cells and the means by which they acquire specific antigen receptors remains one of the central issues in immunology. Abelson virus transformed pre-B cells have been extremely useful for investigating the molecular events controlling immunoglobulin gene rearrangement. Despite the power of this approach, one important feature has been missing from this model: the ability to manipulate the differentiation of the cells in a predictable and controllable fashion. We have overcome this obstacle by using pre-B cells transformed with temperature sensitive Abelson virus. When these cells are incubated at the nonpermissive temperature, light chain gene rearrangement is activated in a high percentage of the cells within one to four days. We will use the temperature sensitive pre-B transformants to ask three questions. 1. Does the pre-B to B cell transition occur in the temperature sensitive transformants and is it regulated by immunoglobulin molecules? The high frequency of light chain rearrangement observed in the temperature sensitive transformants may reflect activation of this single event or activation of the B cell differentiation program. We will examine expression of differentiation markers to distinguish between these two possibilities and test the role of immunoglobulin proteins in stimulating differentiation of the cells. 2. Are rearrangement of kappa and lambda genes and RS sequences regulated or stochastic processes? We will use a series of time course experiments to determine if rearrangements of kappa, lambda and RS are related temporally. The relationship between transcriptional activity, changes in chromatin structure and gene rearrangement will be examined. 3. What genes are upregulated during light chain rearrangements? Differential display will be used to identify sequences that are expressed at high levels when light chain gene rearrangement is activated. A long range goal of these experiments is to identify genes that are important for light chain rearrangement.

免疫球蛋白基因重排是发展的基础 淋巴细胞。 了解这些细胞的产生以及通过 他们获得了特定的抗原受体仍然是中央 免疫学问题。 Abelson病毒转化后B细胞已经 对于调查控制的分子事件非常有用 免疫球蛋白基因重排。 尽管有这种方法的力量， 该模型缺少一个重要功能： 操纵细胞在可预测的和 可控的时尚。 我们通过使用Pre-B细胞克服了这一障碍 用温度敏感的阿伯森病毒转化。 当这些细胞时 在非渗透温度，轻链基因下孵育 重排以高度的单元格中激活 四天。 我们将使用温度敏感的前B变换体问三个 问题。 1。前B到B细胞过渡是否发生在 温度敏感的转化体，是否由免疫球蛋白调节 分子？ 轻链重排的高频在 温度敏感转化体可能反映这一点的激活 单个事件或B细胞分化程序的激活。 我们将 检查分化标记的表达以区分这些标记 两种可能性并测试免疫球蛋白蛋白在 刺激细胞的分化。 2。是Kappa的重排 和lambda基因和RS序列受调节或随机过程？ 我们 将使用一系列的时间课程实验来确定是否是否 Kappa，Lambda和RS的重排在时间上是相关的。 这 转录活性之间的关系，染色质的变化 将检查结构和基因重排。 3。什么是基因 轻链重排期间上调？ 差分显示器将 用于识别光线时在高水平表达的序列 链基因重排被激活。 这些的远距离目标 实验是确定对轻链很重要的基因 重排。