STUDIES OF HLA CLASS II GENES

建议进行研究，以获得有关序列的进一步信息， 表达的HLA II类DR和DQ基因负责 T淋巴细胞识别的决定簇在同种异体移植中的表达 有限的承认。 纯合子分型细胞（HTC），来自 在可能的情况下，将研究近亲交配。 DR 4+和DR 2 + HTCs将是几年内拟议实验的主要焦点。 原因，包括这些中的每一个的几个“Dw亚型” 已经定义了血清学上定义的特异性。 的个人 根据Dw特异性，DR 4+可分为许多组 它们表达的HTCs定义的特异性，即T 淋巴细胞;这些“亚型”特异性包括Dw 4，Dw 10，Dw 13，Dw 14， DW 15。 同样，DR 2+个体可以分为表达DR 2+的个体。 Dw 2、Dw 12和LD-MN 2。 我们认为这是一个特殊的力量， 本文提出的用于通过DNA技术研究的相同HTC的应用 我们的实验室一直在研究，并将继续研究， DR和DQ蛋白的研究， 反应性T淋巴细胞，以及 异源和限制性识别。 这应该允许相关性， 如本申请中的协议所确定的结构， 功能，从而更好地了解分子基础， 细胞相互作用 许多技术，包括体内 重组、外显子内改组和定点诱变， 提出精确地确定哪些序列参与表达 T淋巴细胞识别的决定簇。 正常基因和诱变基因 将通过转染进行研究。 此外，肽，包括 将在抗原呈递细胞中评价感兴趣的序列 系统 将尝试生成单克隆抗体/抗血清 兴趣的决定因素。 我们相信这些研究将有助于 通过提供研究所需的背景信息进行诊断 “疾病单倍型”中的基因家族。

Studies are proposed to obtain further information about sequences of expressed HLA class II DR and DQ genes that are responsible for the expression of determinant recognized by T lymphocytes in allo- or restricted recognition. Homozygous typing cells (HTCs), from consanguineous matings wherever possible, will be studied. DR4+ and DR2+ HTCs will be the primary focus of the proposed experiments for several reasons, including the fact that several "Dw subtypes" of each of these serologically defined specificities have been defined. Individuals who are DR4+ can be divided into a number of groups based on the Dw specificities that they express as defined with HTCs, i.e. specificities recognized by T lymphocytes; these "subtype" specificities include Dw4, Dw10, Dw13, Dw14, and Dw15. Likewise, DR2+ individuals can be divided into those expressing Dw2, Dw12, and LD-MN2. We consider it a special strength of this application that the same HTCs proposed herein for study by DNA techniques have been, and are continuing to be, studied in our laboratories including studies of the DR and DQ proteins, functional characterization of responding T lymphocytes, and definition of determinants recognized in allo- and restricted recognition. This should allow correlations of structure as determined by protocols in the present application with function and thus allow a better understanding of the molecular basis of cellular interactions. A number of techniques, including in vivo recombination, intra-exonic shuffling and site directed mutagenesis, are proposed to pinpoint precisely which sequences are involved in expression of determinants recognized by T lymphocytes. Normal and mutagenized genes will be studied by transfection. In addition, peptides which include sequences of interest will be evaluated in an antigen presenting cell system. Attempts will be made to generate monoclonal antibodies/antisera to determinants of interest. We believe these studies will contribute to diagnosis by providing the background information needed to study this same gene family in "disease haplotypes".