STRUCTURE AND FUNCTION OF THE I REGION GENES

建议的项目旨在研究该结构和 MOSE H-2 I区基因产物的免疫调节作用 很复杂。我们的第一个具体目标是完成该基因的基因组克隆 I地区。重点将是克隆H-2K和I-A之间的基因组DNA 使用粘粒文库和染色体行走技术进行测试。这些 克隆人将被检查是否存在已经被 基因定位到Ia，但还没有蛋白质产品 与世隔绝。I区克隆的完成将提供 关于这个区域编码的基因的数量以及可能的它们的答案 功能。 第二个具体目标将是确定具体的 Ia与T细胞的相互作用。因为辅助性T细胞只能识别 在Ia抗原的背景下，Ia抗原起着至关重要的作用 在这种细胞间的相互作用中扮演着重要的角色。通过结合最近开发的 使用体外诱变技术的转基因程序，我们将改变 将I-A和I-E基因产物导入抗原前的研究 展示细胞，然后测量它们在功能上相互作用的能力 与T细胞有关。 第三个具体目标是确定B细胞上是否表达I-A或I-E 能够向B细胞传递信号，该信号可能部分地 负责它的激活。为了成功回答这个问题，我们 会在小的静息B细胞中表达改变的Ia抗原，并确定 在抗原和辅助性T细胞存在的情况下，B细胞仍然可以被激活 细胞。我们建议使用的传递系统用于在B细胞中表达 是牛痘病毒。

The projects proposed are designed to study the structure and immunoregulatory role of the gene products in the I region of the mose H-2 complex. Our first specific aim is to complete the genomic cloning of the I region. The focus will be to clone the genomic DNA between H-2K and I-A Beta using cosmid libraries and the technique of chromosome walking. These clones will be examined for the presence of genes which have been genetically mapped to Ia but for which no protein product has been isolated. The completion of the cloning of the I region will provide answers as to the number of genes encoded in this region and possibly their function. The second specific aim will be to determine the nature of the specific interaction between Ia and the T cell. Since helper T cells only recognize antigen in the context of the appropriate Ia antigen, Ia plays a crucial role in this cell-cell interaction. By combining recently developed transfection procedures with in vitro mutagenesis techniques, we will alter the gene products for I-A and I-E before introducing them into antigen presenting cells and then measure their ability to functionally interact with T cells. The third specific aim is to determine if I-A or I-E expressed on B cells is capable of delivering a signal to the B cell which may in part be responsible for its activation. To successfully answer this question, we will express altered Ia antigens in small resting B cells and determine if the B cells can still be activated in the presence of antigen and T helper cells. The delivery system we propose to use for the expression in B cells is vaccinia virus.