NON-H-2 HISTOCOMPATIBILITY GENES AND ENCODED ANTIGENS

同种异体组织移植的成败取决于产品 一个常染色体和性连锁的多态基因大家族的成员 指定的组织相容性(H)基因。面临的最大障碍 成功的移植取决于同种异体抗原产物 主要组织相容性复合体(MHC)的I类和II类基因。 MHC I类和II类供受者的配型 同种异体抗原不仅减少或消除了MHC编码的屏障， 相反，优化供体次要H抗原的呈递 要求捐赠者和接受者的MHC匹配才能演示。这个 证明单个次要H抗原的抗原成分是 与MHC分子的多肽结合部位结合的多肽不提供 这不仅是为什么微小的H基因未能被克隆的原因，而且也提供了 一种鉴定和测序这些H抗原的实验方法 多肽。利用所需的I类分子的知识 单个次要H抗原呈递给细胞溶解T淋巴细胞 (CTL)和内源性多肽亚群与个体结合的事实 I类分子，我们有来自于 多个微小的H抗原。这些次要H的氨基酸序列 多肽可用于克隆和测序各自的基因。 这项研究计划的总体目标是克隆和识别 了解小鼠微小H基因的特异性和遗传学 控制T细胞对其编码抗原的反应。第一个目标是 小H抗原肽的氨基酸序列的质谱学研究 以及氨基酸序列在简并化合物合成中的应用 用于克隆微小H基因cDNA的寡核苷酸。H基因的cDNA将 经测序和DNA介导的真核基因转移证实 细胞，随后用特定的CTL进行测试。第二个目标是 近交系小鼠表达的次要H肽的多样性估计 通过扩增和测序编码该基因的DNA序列 同源多肽。第三个目标是检验我们的假设。 次要H肽由一组非丰富的内源肽组成 对I类分子表现出相对更高的亲和力 由丰富的内源性多肽表现出来的。小H来源的多肽 抗原、病毒和可溶性抗原，以及丰富的内源性蛋白 在竞争性结合分析和分析中进行相对亲和力测试 用于稳定I类分子的表达：多肽复合体。第四次 目的是对免疫优势的次要H抗原多肽进行测序。 对这样一种假设的检验：这些优势抗原的排名 是基于它们的多肽对I类分子的相对亲和力。 这些研究构成了T细胞对Minor反应的第一次分析 将对次要H抗原肽和基因进行测序的H抗原 以及研究这些结合的亲和力和特异性 多肽转化为MHC I类分子。

The success or failure of tissue allografts is determined by the products of a large family of autosomal and sex-linked, polymorphic genes that are designated histocompatibility (H) genes. The strongest barriers to successful transplantation are determined by the alloantigenic products of class I and class II genes of the major histocompatibility complex (MHC). The matching of donors and recipients for MHC class I and class II alloantigens not only reduces or eliminates MHC-encoded barriers but, conversely, optimizes the presentation of donor minor H antigens that require MHC matching of donor and recipient for presentation. The demonstration that the antigenic components of single minor H antigens are peptides bound to the peptide binding site of MHC molecules provides not only a reason for why minor H genes have eluded cloning, but, also provides an experimental approach to identifying and sequencing these H antigen peptides. Using the knowledge of the class I molecule that is required for the presentation of a single minor H antigen to cytolytic T lymphocytes (CTL) and the fact that subsets of endogenous peptides bind to individual class I molecules, we have chromatographically resolved peptides from multiple minor H antigens. The amino acid sequences of these minor H peptides can be applied to cloning and sequencing the respective genes. The overall objectives of this research program are to clone and identify minor H genes in the mouse and to understand the specificity and genetic control of the T cell response to their encoded antigens. The first aim is the amino acid sequencing of minor H antigen peptides by mass spectrometry and application of the amino acid sequences to the synthesis of degenerate oligonucleotides for the cloning of minor H gene CDNAS. H gene CDNAS will be confirmed by sequencing and DNA-mediated gene transfer into eukaryotic cells with subsequent testing with specific CTL. The second objective is the estimation of diversity of minor H peptides expressed by inbred mouse strains by amplification and sequencing of DNA sequences that encode the homologous peptides. The third objective is the testing of our hypothesis that minor H peptides comprise a group of non-abundant endogenous peptides that exhibit relatively higher affinity for class I molecules than that exhibited by abundant, endogenous peptides. Peptides derived from minor H antigens, viral and soluble antigens, and abundant endogenous proteins will be tested for relative affinity in competitive binding assays and assays for stabilization of expression of class I:peptide complexes. The fourth objective is the sequencing of immunodominant minor H antigen peptides and the testing of the hypothesis that the ranking of these dominant antigens is based on the relative affinity of their peptides for class I molecules. These studies constitute the first analysis of the T cell response to minor H antigens that will both sequence minor H antigen peptides and genes as well as investigate the affinity and specificity of the binding of these peptides to MHC class I molecules.