STRUCTURE AND FUNCTION OF THE I REGION GENES

拟议的项目旨在研究结构， 小鼠H-2 Ⅰ区基因产物的免疫调节作用 复杂. 我们的第一个具体目标是完成基因组克隆的 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步移技术的Beta。 这些 将检查克隆中存在的基因， 基因定位于Ia，但没有蛋白质产物被 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 功能 第二个具体目标将是确定具体的 Ia和T细胞之间的相互作用。 因为辅助性T细胞只识别 抗原在适当的Ia抗原的背景下，Ia起着至关重要的作用。 在细胞间相互作用中的作用。 通过结合最近开发的 转染程序与体外诱变技术，我们将改变 在将I-A和I-E的基因产物引入抗原之前， 然后测量它们在功能上相互作用的能力， T细胞。 第三个具体目的是确定I-A或I-E是否在B细胞上表达 能够向B细胞递送信号，该信号可以部分地 负责其激活。 为了成功回答这个问题，我们 将在小的静息B细胞中表达改变的Ia抗原，并确定 B细胞在抗原和辅助性T细胞存在下仍能被激活 细胞 我们建议用于在B细胞中表达的递送系统 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.