ANTIGEN RECEPTOR GENES FROM GAT-SPECIFIC TH AND TS CELLS

该项目的长期目标是描述 由鼠B淋巴细胞和T淋巴细胞家族使用的抗原受体 参与对单一抗原的免疫应答的淋巴细胞。 我们将 从两种辅助（TH）细胞中分离编码抗原结合多肽基因， 和对多肽抗原GAT具有特异性的抑制性（TS）T细胞。 已经克隆了几类GAT特异性淋巴细胞，包括B 主要功能是合成GAT结合免疫球蛋白的细胞 (Ig)辅助性T细胞（TH），其功能是增加GAT-Ig的产生，以及 两个抑制性T细胞亚群（TS1和TS2）， 回应GAT。 编码B细胞GAT-Ig的基因已被分离 以前。 从GAT-TS细胞中克隆受体基因， 将用于cDNA文库的构建。 cDNA将是 克隆到表达载体LambdagtII中，并用1） TS上I-J、独特型和大津/Tind决定簇的血清学试剂 细胞，2）与预测的mRNA互补的寡核苷酸探针， 分泌的抗原结合抑制因子的序列，和3） 富集TS特异性转录物的消减探针。 分离的cdna 然后将使用以下方法确认仅在T细胞中转录的克隆： 功能标准。 从GAT-TH细胞中分离受体基因 从基因组文库中通过与分离自 非GAT-TH细胞。 T细胞受体基因将用于表征可能的DNA 重组，基因组组织，以及产生 多样性 TS抗原编码基因的染色体定位 受体和I-J，独特型，和大津/Tind决定因素将是 使用小鼠-仓鼠杂交系确定。 同样，受体基因 由几种不同的TH细胞使用，这些TH细胞结合GAT，但它们的MHC不同 限制，将进行比较。 关于监管的初步研究 使用各种抗原和免疫抑制剂在TH细胞中转录受体基因， 还描述了细胞刺激。 通过比较基因序列 被各种淋巴细胞类用来结合GAT结合多肽， 我们将处于一个独特的位置来研究抗原的分子基础 识别、MHC限制、免疫细胞相互作用以及T和B 淋巴细胞活化 更好地理解分子控制 在免疫系统中起作用是决定 免疫疾病状态的基本机制和缺陷。

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.