ANTIGEN RECEPTOR GENES FROM GAT-SPECIFIC TH AND TS CELLS

来自 GAT 特异性 TH 和 TS 细胞的抗原受体基因

• 批准号: 3132973
• 负责人: ELLEN KRAIG
• 金额: $ 9.24万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-04-01 至 1988-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3132973
• 关键词: B lymphocyte; T lymphocyte; complementary DNA; genetic library; helper T lymphocyte; hybridomas; immune response genes; immunogenetics; immunoglobulin genes; lac operon; leukocyte antigen typing; major histocompatibility complex; molecular cloning; nucleic acid sequence; suppressor T lymphocyte; surface antigens

The long-term goal of this project is the characterization of the antigen-receptors used by the family of murine B lymphocytes and T lymphocytes involved in the immune response to a single antigen. We will isolate genes encoding antigen-binding polypeptides from both helper (TH) and suppressor (TS) T cells specific for the polypeptide antigen, GAT. Several classes of GAT-specific lymphocytes have been cloned, including B cells whose primary function is the synthesis of GAT-binding immunoglobulin (Ig), helper T cells (TH) that function to augment GAT-Ig production, and two subsets of suppressor T cells (TS1 and TS2) that diminish the immune response to GAT. The genes encoding B-cell GAT-Ig have been isolated previously. To clone the receptor genes from GAT-TS cells, TS1 and TS2 RNA will be used in the construction of cDNA libraries. The cDNA will be cloned into Lambdagtll, an expression vector, and screened with 1) serological reagents to the I-J, idiotypic, and Tsu/Tind determinants on TS cells, 2) oligonucleotide probes complementary to the mRNA predicted from the sequences of secreted antigen-binding suppressive factors, and 3) subtractive probes enriched for TS-specific transcripts. Isolated cDNA clones that are transcribed only in T cells will then be confirmed using functional criteria. The receptor genes from GAT-TH cells will be isolated from a genomic library by hybridization to a TH-probe isolated from non-GAT-TH cells. The T cell receptor genes will be used to characterize possible DNA rearrangements, the genomic organization, and the mechanisms for generating diversity. The chromosomal locations of the genes encoding TS antigen receptors and the I-J, idiotypic, and Tsu/Tind determinants will be ascertained using mouse-hamster hybrid lines. Also, the receptor genes used by several different TH cells that bind GAT, but differ in their MHC restriction, will be compared. Initial studies of the regulation of transcription of the receptor genes in TH cells using various antigenic and cellular stimuli are also described. By comparing the genetic sequences used by the various lymphocyte classes to enclode GAT-binding polypeptides, we will be in a unique position to study the molecular bases of antigen recognition, MHC restriction, immune cellular interactions, and T and B lymphocyte activation. A better understanding of the molecular controls operating in the immune system is of primary importance in determining basic mechanisms and defects in immune disease states.

这个项目的长期目标是描述 小鼠B淋巴细胞和T淋巴细胞家族使用的抗原受体 淋巴细胞参与对单一抗原的免疫反应。我们会 从两个辅助者(TH)中分离抗原结合多肽编码基因 和多肽抗原GAT特异性的抑制(TS)T细胞。 已经克隆了几类GAT特异的淋巴细胞，包括B细胞 主要功能是合成GAT结合免疫球蛋白的细胞 (IG)，辅助T细胞(TH)，其功能是增加GAT-Ig的产生，以及 抑制T细胞的两个亚群(TS1和TS2)，它们会降低免疫 对关税及服务税的回应。编码B细胞GAT-Ig的基因已被分离 之前。克隆GAT-TS细胞受体基因TS1和TS2RNA 将用于构建cDNA文库。该基因将被命名为 克隆入表达载体Lambdagtll，经1)筛选 针对TS的I-J、独特型和Tsu/Tind决定簇的血清学试剂 细胞，2)寡核苷酸探针，互补于从 分泌型抗原结合抑制因子的序列；3) 富含TS特异性转录本的消减探针。分离的c DNA 只在T细胞中转录的克隆将被确认为使用 功能标准。GAT-TH细胞的受体基因将被分离 通过杂交从基因组文库中分离出TH探针 非GAT-TH细胞。 T细胞受体基因将用于表征可能的DNA 重排、基因组组织和生成的机制 多样性。TS抗原编码基因的染色体定位 受体和I-J、独特型和Tsu/Tind决定因素将是 利用小鼠-仓鼠杂交品系进行了确认。此外，受体基因 由几个不同的TH细胞使用，这些细胞与GAT结合，但它们的MHC不同 限制，将进行比较。关于监管的初步研究 用不同的抗原性和特异性转录TH细胞中的受体基因 还描述了细胞刺激。通过比较基因序列 被不同类别的淋巴细胞用来包裹GAT结合多肽， 我们将在一个独特的位置研究抗原的分子基础 识别、MHC限制、免疫细胞相互作用与T和B 淋巴细胞活化。更好地理解分子控制 在免疫系统中运作是决定 免疫疾病状态的基本机制和缺陷。