EXPRESSION OF SURFACE MARKERS ON T CELLS IN TRANSGENIC MOUSE MODEL

转基因小鼠模型中 T 细胞表面标记的表达

• 批准号: 6099162
• 负责人: HUGH O MCDEVITT
• 金额: $ 13.45万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6099162
• 关键词: CD4 molecule; T cell receptor; animal breeding; antibody formation; biological models; cell differentiation; experimental allergic encephalomyelitis; flow cytometry; gene expression; genetic markers; genetic promoter element; genetically modified animals; helper T lymphocyte; human genetic material tag; insulin dependent diabetes mellitus; interferon gamma; interleukin 4; laboratory mouse; leukocyte activation /transformation; model design /development; monoclonal antibody; natural killer cells; reporter genes; surface antigens; tissue /cell culture

Transgenic mice will be produced which express human Thy-1 under the control of the murine IFN-gamma promoter and murine Thy-1.1 under the control of the murine IL-4 promoter. These transgenes will be introduced into a number of strains which are murine Thy-1.2+. Because of the promoters used, human Thy-1 will be expressed in TH 1 T cells expressing IFN-gamma and murine Thy-1.1 will be expressed in TH2 cells expressing IL- 4. It is possible that both human Thy-1 and murine Thy-1.1 will be expressed in THO cells. This double positivity will then be a marker for the THO subset. Utilizing the appropriate monoclonal antibodies, these transgenic mice will provide unique cell surface markers for the detection of the TH1, THO and TH2 T cell subsets.(monoclonal antibodies for CD8 and NK1 will be required to detect the expected small number of CD8+ T cells and NK cells which also express IFN-gamma). These reporter gene transgenes will then be introduced into the BALB/c, B10.D2, NOD, NOD.A-beta-d, NOD.A-beta-g7.PD, B10.PL, NZB. MRL inbred strain backgrounds. The transgenes will also be introduced into three T cell receptor transgenic lines expressing T cell receptor specific for a myelin basic protein peptide and two influenza hemagglutinin peptides recognized by a CD4 and a CD8 T cell receptor. The presence of the reporter gene transgenes in these inbred strains will permit an analysis of the role of the TH1, THO, and TH2 T cell subsets in the normal immune response in a variety of immunizing conditions, and in the autoimmunity seen in the NOD mouse, in experimental allergic encephalomyelitis (EAE) in the (B10.PL x SJL)F1 mouse, and in the autoimmune syndromes seen in the NZB x NZW mouse and the MRL-lpr/lpr mouse. In a similar manner, the transgenes will permit the analysis of TH1 and TH2 T cell subsets in transgenic mice injected with the peptide for which the T cell receptor is transgenic and will permit a study of the relative inducibility of apoptosis anergy and deletion in the TH l and TH2 cell subsets. In addition, these reporter gene/transgenic mouse lines will permit an analysis of the role of T cells subsets in a wide variety of immune conditions and will be made available to other investigators in this program project as well as to other investigators in this and other institutions. Ultimately, these transgenic mice offer the possibility of validating the very productive paradigm for T cell subset differentiation initially proposed by Coffmann and Mossmann.

将产生转基因小鼠，在 控制鼠的IFN-GAMMA启动子和鼠THY-1.1 对鼠IL-4启动子的控制。这些转基因将被引入 成许多菌株，这些菌株是Thy-1.2+的菌株。因为 使用的启动子，人THY-1将在表达的1个T细胞中表达 IFN-Gamma和Murine Thy-1.1将在表达IL-的Th2细胞中表达 4。 在细胞中表达。 然后，这种双重阳性将成为 Tho子集。 利用适当的单克隆抗体，这些 转基因小鼠将为检测提供独特的细胞表面标记 Th1，Tho和Th2 T细胞子集的（CD8和 将需要NK1检测预期的少量CD8+ T细胞 以及也表达IFN-gamma的NK细胞）。 然后，这些记者基因转基因将被引入BALB/C中 b10.d2，点头，点头。 Mrl Inbred 应变背景。 转基因也将引入三个T 细胞受体转基因线，表达特异性A的T细胞受体 髓磷脂碱性蛋白质肽和两种流感血凝素肽 由CD4和CD8 T细胞受体识别。 存在 这些近交菌株中的记者基因转基因将允许分析 正常免疫中Th1，ThO和Th2 T细胞子集的作用 在各种免疫条件下以及自身免疫性的反应 在点头小鼠中，在实验过敏性脑脊髓炎（EAE）中可见 （b10.pl x sjl）F1鼠标，以及在自身免疫性综合征中 NZB X NZW小鼠和MRL-LPR/LPR鼠标。 以类似的方式 转基因将允许对Th1和Th2 T细胞子集进行分析 转基因小鼠注射了T细胞受体为的肽 转基因，将允许研究 Th L和Th2细胞子集中的凋亡厌氧和缺失。在 此外，这些记者基因/转基因小鼠系将允许 分析T细胞在多种免疫中的作用 条件并将提供给其他调查人员 计划项目以及其他研究人员和其他研究人员 机构。 最终，这些转基因小鼠提供了可能性 验证T细胞子集分化的非常有效的范式 最初由科夫曼和莫斯曼提出。