IN VIVO ROLE OF NF-KB/REL FACTORS IN IMMUNE RESPONSES

描述(改编自调查人员摘要)：NF-kappaB/REL 转录因子是一个由五种蛋白质组成的家族，其功能是二聚体。 复合体参与调节基因的先天和特异性 豁免权。这项研究将研究这些转录因子是如何调节 体内免疫反应。我们的实验方法将利用 针对单个家庭成员的基因靶向小鼠与两种 独特的实验方法。第一种方法是用组织化学方法 直接监测体内核因子-kappaB/Rel活化的标志物。这个记号笔 是通过有针对性地将LacZ报告基因导入 I-kappa-B-α基因座。第二种方法使用标记的逆转录病毒载体 用绿色荧光蛋白重建原代B细胞 特定形式的核因子-kappaB/Rel蛋白。逆转录病毒重建将是 用于确定单个复合体如何调节班级切换和 B细胞分泌免疫球蛋白。定义自然生理学 这些因素的作用将直接有助于理解为什么某些 成员是原癌基因，以及它们的失调如何导致 人类癌症中的致癌激活。决定这些因素如何 调节免疫反应也将使我们更好地了解 临床相关药物的体内作用，如阿司匹林和 糖皮质激素，已被确定为这种抑制物 转录因子家族。具体目标1将决定何时何地 核因子-kappaB/Rel复合体在体内被激活 对T细胞依赖抗原的免疫反应。特定目标2将 在体内研究单个核因子-kappaB/Rel因子的缺失如何扰乱 有效的免疫反应和确定缺陷的细胞基础 回应。目标3将定义特定的核因子-kappaB/REL复合体在 初级B细胞在调节免疫球蛋白分泌和类转换中的作用。

DESCRIPTION (Adapted from the Investigator's abstract): NF-kappaB/Rel transcription factors are a family of five proteins which function as dimer complexes in the regulation of genes involved in innate and specific immunity. This study will examine how these transcription factors regulate in vivo immune responses. Our experimental approach will utilize gene-targeted mice for individual family members in combination with two unique experimental approaches. The first approach uses a histochemical marker that directly monitors in vivo NF-kappaB/Rel activation. This marker was created by targeted introduction of a lacZ reporter gene into the I-kappa-B-alpha locus. The second approach uses retroviral vectors tagged with a green fluorescent protein to reconstitute primary B-cells with specific forms of NF-kappaB/Rel proteins. Retroviral reconstitution will be used to determine how individual complexes regulate class switching and immunoglobulin secretion in B-cells. Defining the natural physiological roles of these factors will directly contribute to understanding why certain members are proto-oncogenes, and how their disregulation can lead to oncogenic activation in human cancer. Determining how these factors regulate immune responses will also produce a better understanding of the in vivo action of clinically relevant drugs, such as aspirin and glucocorticoids, that have been identified as inhibitors of this transcription factor family. Specific Aim 1 will determine when and where NF-kappaB/Rel complexes are activated in vivo during the natural history of an immune response to a T-cell dependent antigen. Specific Aim 2 will examine in vivo how the absence of individual NF-kappaB/Rel factors disrupts effective immune responses and determine the cellular basis for defective responses. Aim 3 will define roles of specific NF-kappaB/Rel complexes in primary B-cells in regulating immunoglobulin secretion and class switching.