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

我们建议的目标是研究与 具有MHC扩展单倍型。我们将检验两种不同的假设 与其职能相关的： (1)某些MHC单倍型纯合子个体与 对乙肝表面抗原无反应或无反应可能是由于TH1或 Th2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)在有应答者和无应答者中。 我们将使用相关和不相关的人类白细胞抗原完全相同的对。 (2)我们将检验以下假设：NK活性低的个体和 低CD56阳性细胞数属于不同的人类白细胞抗原-B、C互补 对应于人类白细胞抗原B、C纯合子和特定人类白细胞抗原B、C的组 杂合组合。我们将生产NK同种异体反应克隆 对一组EBV转化的细胞中的部分细胞而不是全部细胞起反应 不同基因类型的品系或PHA突变体。具体目标是：1. 扩展NK互补基团的定义和特征 基因。我们将研究NK活性和NK(CD16+CD56+)细胞的数量。 以下各项的纯合子或杂合子个体 决定因素：人类白细胞抗原B7(Cw7)、人类白细胞抗原B8(Cw7)、人类白细胞抗原B44(Cw4)、B44(Cw5)、 B57(CW6)、HLAB35(CW4)、HLAB60(Cw3)、HLAB62(Cw3)和HLAB18(CW5)， 人类白细胞抗原B18(Cw3)和人类白细胞抗原B51(Cwx)。二、为了识别NK 特异性并确定它们与扩展单倍型的关联 或携带人类白细胞抗原-B(C)等位基因 将产生并鉴定针对[人类白细胞抗原-B8， CS01，DR3]，[HLA-B18，F1C30，DR3][HLA-B7，SC31，DR2]，[HLA-B62，SC33， DR4]，[HLA-B35，SC31，DR4]纯合扩展单倍型和抗 人类白细胞抗原-B51纯合子个体及其细胞毒活性检测 对抗一组PHA激活的T细胞或EBV转化的B细胞留置权。 我们还将用带有单个等位基因的转染体来测试这些克隆 人类白细胞抗原-C和人类白细胞抗原B基因。我们将使用人类白细胞抗原重组家族(人类白细胞抗原- A/C，B；C/B或B/DR)和不同人类白细胞抗原的纯合子和杂合子细胞 定位NK靶基因表达调控基因的基因分型 特异性AIM II产生的克隆识别的抗原和基因 控制NK细胞的数量和活性。方法论将使用 自然杀伤细胞的克隆及其免疫荧光法鉴定 检测，聚合酶链式反应检测人类白细胞抗原-C分型，并转染类 具有人类白细胞抗原I类基因的阴性细胞和诱变作用。身份识别 人类的造血组织相容性(HN)遗传系统可能是 适用于一次骨髓移植的配型。

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.