KAPPA GENE REARRANGEMENT IN TRANSFORMED PREB CELLS

免疫球蛋白基因重排是发展免疫球蛋白的基础。 淋巴细胞 了解这些细胞的产生和方法， 它们获得的特异性抗原受体仍然是 免疫学的问题。 Abelson病毒转化的前B细胞已经被 对于研究控制 免疫球蛋白基因重排 尽管这种方法很强大， 这种模式缺少一个重要特征： 以可预测的方式操纵细胞的分化， 可控制的方式。 我们已经通过使用前B细胞克服了这一障碍 用温度敏感的Abelson病毒转化。 当这些细胞 在非允许温度下孵育，轻链基因 重排在一个细胞内的高百分比中被激活， 四天了 我们将使用温度敏感的前B转化体来询问三个 问题. 1.前B细胞到B细胞的转变是否发生在 温度敏感的转化体，它是由免疫球蛋白调节 分子？ 观察到的高频率轻链重排， 温度敏感性转化体可以反映这种激活 单个事件或B细胞分化程序的激活。 我们将 检测分化标志物的表达以区分这些 两种可能性，并测试免疫球蛋白的作用， 刺激细胞的分化。 2. Kappa重排 和λ基因和RS序列调节或随机过程？ 我们 将使用一系列时间过程实验来确定 κ、λ和RS的重排在时间上是相关的。 的 转录活性、染色质变化 结构和基因重排将被检查。 3.什么基因是 在轻链重排过程中上调？ 差分显示将 用于鉴定当光照时以高水平表达的序列， 链基因重排被激活。 一个长期的目标， 实验的目的是确定对轻链重要的基因， 重排

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.