PROCESSING OF ANTIGENS FOR HLA RESTRICTED PRESENTATION

这一应用的长期目标是阐明 在抗原呈递中产生MHC分子/肽复合体 手机。这项建议的具体目的是在功能上和 一组突变的抗原提呈细胞的遗传学特征，它 不能提呈多种可溶性抗原给二类受限 T细胞，但可以呈递一种多肽抗原。方法是：使用 抗原和DR限制性肽特异性T细胞的蛋白水解酶 克隆以更广泛地评估突变体的呈现能力 抗原；以确定突变体是否在抗原上有缺陷 通过内吞途径进行加工；使用体细胞杂交来 确定8个义务中有多少个互补组 独立呈现缺陷突变体；以确定人类是否 非淋巴样细胞和小鼠淋巴样细胞可以弥补缺陷(S) 在突变体中；确定受影响的基因(S)是否映射到X 染色体或6号染色体的HLA-D区；并分离 利用逆转录病毒插入突变的表型相似突变体 最终克隆突变基因的工具(S)。此外，孤立 其他抗原加工突变体与抗原的免疫选择 将追求特定的细胞溶解T细胞。这些研究可能会导致 更好地了解MHC限制性抗原处理/提呈和 从而对人类白细胞抗原相关的病理生理学有了深入的了解。 MHC限制性T细胞可能在疾病中发挥作用。

The long term objective of this application is to elucidate pathways for generating an MHC molecule/peptide complex within an antigen presenting cell. The specific aims of this proposal are to functionally and genetically characterize a group of mutant antigen presenting cells, which are unable to present a variety of soluble antigens to class II restricted T cells but can present a peptide antigen. The approaches are: to use proteolytic digests of antigen and DR restricted peptide specific T cell clones to more extensively evaluate the mutants' ability to present antigen; to determine whether the mutants are defective in antigen processing via the endocytic pathway; to use somatic cell hybrids to determine how many complementation groups exist among 8 obligate independent presentation defective mutants; to determine whether human non-lymphoid cells and murine lymphoid cells can complement the defect(s) in the mutants; to determine whether the affected gene(s) map to X chromosome or the HLA-D region of chromosome 6; and to isolate phenotypically similar mutants using retroviral insertional mutagenesis as a tool for eventual cloning of the mutant gene(s). In addition, isolation of other antigen processing mutants by immunoselection with antigen specific cytolytic T cells will be pursued. These studies may lead to a better understanding of MHC restricted antigen processing/presentation and may thus provide insights into the pathophysiology of HLA associated diseases in which MHC restricted T cells are likely to play a role.