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

NK 和 T 细胞与主要组织相容性复合体相关的研究

• 批准号: 6272668
• 负责人: EDMOND J YUNIS
• 金额: $ 42.95万
• 依托单位: IMMUNE DISEASE INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6272668
• 关键词: MHC class I antigen; T lymphocyte; antigen presenting cell; blocking antibody; family genetics; gene frequency; genetic mapping; genetic polymorphism; genotype; hepatitis B antigens; histocompatibility typing; human subject; immune response genes; immunogenetics; leukocyte activation /transformation; lymphocyte proliferation; major histocompatibility complex; natural killer cells; site directed mutagenesis

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.

我们建议的目的是研究相关的免疫功能 具有 MHC 扩展单倍型。 我们将测试两个不同的假设 与其功能相关： (1) 某些 MHC 单倍型纯合的个体与 对 HBsAG 无反应和无反应可能是由于 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、 在 HBsAg 存在的情况下，应答者和无应答者中存在 IL-10 和 IL-12）。 我们将使用 HLA 相同对，相关的和不相关的。 (2) 我们将检验以下假设：NK 活性低的个体和 CD56阳性细胞数低，属于不同HLA-B、C互补 对应于 HLA-B、C 纯合子和特定 HLA-B、C 的组 杂合组合。 我们将生产 NK 同种异体反应性克隆 对一组 EBV 转化细胞中的部分但不是全部细胞发生反应 不同基因型的品系或 PHA 母细胞。 具体目标是： 1. 扩展 NK 互补基团的定义和表征 基因。 我们将研究 NK 活性和 NK(CD16+CD56+) 细胞的数量 以下各项的纯合或杂合个体 决定簇：HLA-B7 (Cw7)、HLA-B8 (Cw7)、HLA-B44 (Cw4)、B44 (Cw 5)、HLA- B57 (Cw6)、HLA-B35 (Cw4)、HLA-B60 (Cw3)、HLA-B62 (Cw3) 和 HLA-B18 (Cw5)， HLA-B18 (Cw3) 和 HLA-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 或具有不同组成型的个体中的 HLA-B(C) 等位基因，我们 将针对 [HLA-B8、 CS01，DR3]，[HLA-B18，F1C30，DR3] [HLA-B7，SC31，DR2]，[HLA-B62，SC33， DR4]、[HLA-B35、SC31、DR4] 纯合扩展单倍型并针对 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 细胞留置权。 我们还将使用具有单个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- 空调、空调； C/B 或 B/DR) 以及不同 HLA 的纯合和杂合细胞 绘制调节 NK 靶标表达的基因的基因型 由特定目标 II 和基因中生成的克隆识别的抗原 控制 NK 细胞的数量和活性。 将使用方法论 NK 细胞的克隆，通过免疫荧光表征 NK 克隆 检测、聚合酶链式反应 HLA-C 分型以及类转染 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 对于单次骨髓移植的配型很有用。