STUDIES OF NK AND T CELLS IN RELATION TO THE MAJOR HISTOCOMPATIBILITY COMPLEX

我们的目标是研究免疫功能相关的 MHC扩展单倍型。 我们将检验两种不同的假设 与其职能有关： (1)某些MHC单倍型的纯合子个体与 对HLAG无应答和无应答可能是由于TH 1缺陷或 Th 2细胞 我们将使用T细胞前体频率，细胞混合物 细胞因子（干扰素-γ，IL-2，IL-4， IL-10和IL-12）。 我们将使用HLA相同对，相关和无关。 (2)我们将检验一个假设，即NK活性低的个体， 低CD 56阳性细胞数属于HLA-B、C互补型 对应于HLA-B、C纯合子和特异性HLA-B、C的组 杂合组合 我们将产生NK同种异体反应克隆， 对一组EBV转化细胞中的一些但不是所有细胞反应 株系或PHA胚细胞。 具体目标是：1。 到 扩展了NK互补群的定义和特征 基因. 我们将研究NK活性和NK（CD 16 + CD 56+）细胞的数量， 个体对于以下每一个是纯合的或杂合的 决定簇：HLA-B7（Cw 7）、HLA-B8（Cw 7）、HLA-B44（Cw 4）、B44（Cw 5）、HLA-B44（Cw 7）、HLA-B44（Cw 8） B57（Cw6）、HLA-B35（Cw4）、HLA-B60（Cw3）、HLA-B62（Cw3）和HLA-B18（Cw5）， HLA-B18（Cw 3）和HLA-B51（Cwx）。 二. 为了识别NK 特异性，并确定其与扩展单倍型的关联 或与HLA-B（C）等位基因在个体与不同的补体型， 将产生并表征针对[HLA-B8， CS01，DR3]，[HLA-B18，F1C30，DR3]，[HLA-B7，SC31，DR2]，[HLA-B62，SC33， DR 4]，[HLA-B35，SC 31，DR 4]纯合扩展单倍型和抗 HLA-B51纯合个体，并测试其细胞毒活性 针对一组PHA激活的T细胞或EBV转化的B细胞系。 我们还将用具有个别等位基因的转染子测试这些克隆 HLA-C和HLA-B基因。 三. 我们将使用HLA重组家族（HLA- A/C、B; C/B或B/DR）以及不同HLA的纯合和杂合细胞 基因型，以定位调节NK靶标表达的基因 特异性目标II和基因中产生的克隆识别的抗原 控制NK细胞的数量和活性。 方法将使用 NK细胞的克隆，通过免疫荧光表征NK克隆 检测，通过聚合酶链反应进行HLA-C分型，并转染类 具有HLA I类基因和诱变的I阴性细胞。 鉴定 人类的造血组织相容性（Hn）遗传系统可能是 可用于一次骨髓移植中的匹配。

The objective of our proposal is to study immunologic functions associated with MHC extended haplotypes. We will test two different hypotheses related to their functions: (1) Individuals homozygous for certain MHC haplotypes are associated with nonresponse against HBsAG and nonresponse may be due to a defect of TH1 or Th2 cells. We will use T cell precursor frequency, cell mixture experiments and measurements of cytokines (interferon-gamma, IL-2, IL-4, IL-10 and IL-12) in responders and nonresponders in the presence of HBsAg. We will use HLA identical pairs, related and unrelated. (2) We will test the hypothesis that individuals with low NK activity and low CD56 positive cell number belong to different HLA-B, C complementation groups corresponding to HLA-B, C homozygous and specific HLA-B, C heterozygous combinations. We will produce NK alloreactive clones that react against some but not all cells of a panel of EBV-transformed cell lines or PHA blasts of different genotypes. Specific aims are: 1. To extend the definition and characterization of complementation groups of NK genes. We will study NK activity an the number of NK(CD16+CD56+) cells in individuals homozygous or heterozygous for each of the following determinants: HLA-B7 (Cw7), HLA-B8 (Cw7), HLA-B44 (Cw4), B44 (Cw 5), HLA- B57 (Cw6), HLA-B35 (Cw4), HLA-B60 (Cw3), HLA-B62 (Cw3) and HLA-B18 (Cw5), HLA-B18 (Cw3) and HLA-B51 (Cwx). II. In order to identify NK specificities and to determine their association with extended haplotypes or with HLA-B(C) alleles in individuals with differences in complotypes we will generate and characterize alloreactive NK clones against [HLA-B8, CS01, DR3], [HLA-B18, F1C30, DR3] [HLA-B7, SC31, DR2], [HLA-B62, SC33, DR4], [HLA-B35, SC31, DR4] homozygous extended haplotypes and against individuals homozygous for HLA-B51 and test their cytotoxic activity against a panel of PHA-activated T cells or EBV-transformed B cell liens. We will also test these clones with transfectants with individual alleles of HLA-C and HLA-B genes. III. We will use HLA-recombinant families (HLA- A/C, B; C/B or B/DR) and homozygous and heterozygous cells of different HLA genotypes to map the gene regulating the expression of the NK target antigens recognized by clones generated in specific aim II and the gene controlling the number and activity of NK cells. Methodologies will use cloning of NK cells, characterization of NK clones by immunofluorescence assay, HLA-C typing by polymerase chain reaction, and transfection of class I negative cells with HLA class I genes and mutagenesis. Identification of a hematopoietic histocompatibility (Hn) genetic system in humans could be useful for matching in one marrow transplantation.