TRANSGENIC MOUSE MODEL FOR TOLERANCE IN THE EYE

眼部耐受性转基因小鼠模型

• 批准号: 2163315
• 负责人: Jerold G Woodward
• 金额: $ 21.87万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-12-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2163315
• 关键词: MHC class I antigen; MHC class II antigen; T lymphocyte; age difference; autoimmune disorder; crystallins; disease /disorder model; flow cytometry; fusion gene; gene expression; genetically modified animals; histocompatibility antigens; immune tolerance /unresponsiveness; immunocytochemistry; immunogenetics; inflammation; interferon gamma; isoantigen; laboratory mouse; lens; model design /development; monoclonal antibody; tissue /cell culture; uveitis

A central question in the understanding of autoimmune disease processes is how T cell tolerance is developed and maintained to self antigens. Tolerance to self-antigens is acquired in the thymus through a process termed negative selection, and in the allogeneic MHC or foreign antigens are expressed only in extrathymic tissues have provided considerable insight into the mechanisms of peripheral tolerance induction, but still many questions remain. We have developed a new transgenic model for tolerance in which an allogeneic mcH antigen or IFn-gamma is expressed in the lens of the eyes, an immunologically privileged site. The lens of the eye represents and excellent model for further study of peripheral tolerance for several reasons. It lacks vascularization and lymphatic drainage Thus, this site may have unique properties concerning tolerance induction. The eye is also a prominent target in several autoimmune diseases including HLA-B27 linked Reiter's syndrome. In order to study tolerance induction to antigens expressed in the lens, transgenic mice have been produced expressing MHC class I (H-2D) or interferon gamma under expression and appear to be promising models for tolerance studies. These models will be immunohistochemistry. Experiments will also determine whether the H-2D antigen expressed in the lens is associated with Beta 2-microglobulin. The inflammatory changes taking place in these mice will be carefully evaluated histologically at different ages. Experiments are also proposed to test whether these changes are immunologically mediated. For the class I transgenic mice, we will assess the degree of tolerance to the allo class I transgenic in both in vitro and in vivo systems. For the INF-gamma transgenic mice, we will determine whether specific T cell sensitization to ocular antigens is occurring and whether IFN-gamma expression in the eye modifies the immunosuppressive nature of the eye. Finally, we propose to produce a third model by expressing I-E antigens under the control of the alpha-A crystallin promoter. In these mice, clonal deletion of Vbeta17a (I-E reactive) T cells will be assessed by monoclonal antibody staining and flow cytometry. These studies will provide important new information on the mechanisms of tolerance induction to tissue restricted antigens, and provide new mouse models for autoimmune uveitis.

了解自身免疫性疾病过程的一个中心问题 T细胞对自身抗原的耐受性是如何形成和维持的。 对自身抗原的耐受性是通过一个过程在胸腺中获得的。 在同种异体MHC或外源抗原中， 仅在胸腺外组织中表达， 深入了解外周耐受诱导的机制，但仍然 许多问题仍然存在。 我们开发了一种新的转基因模型， 表达同种异体mcH抗原或IFN-γ的耐受性 在眼睛的透镜中，一个免疫特权部位。 该透镜 的眼睛代表和良好的模型，为进一步研究周边 宽容有几个原因。 它缺乏血管和淋巴管 因此，这个网站可能有独特的属性有关的宽容 诱导 眼睛也是几种自身免疫性疾病的突出目标。 包括HLA-B27相关的赖特综合征。 为了研究 对透镜中表达的抗原的耐受诱导，转基因小鼠 已经产生表达MHC I类（H-2D）或干扰素γ的 在表达下，似乎是耐受性研究的有前途的模型。 这些模型将进行免疫组织化学。 实验也将 确定透镜中表达的H-2D抗原是否相关 β 2微球蛋白 炎症变化发生在 将在不同年龄对这些小鼠进行仔细的组织学评价。 实验也提出了测试这些变化是否是 免疫介导的 对于I类转基因小鼠，我们将 评估在两种情况下对异源I类转基因的耐受程度， 体外和体内系统。 对于INF-γ转基因小鼠， 确定是否特异性T细胞致敏眼抗原， 发生以及眼睛中IFN-γ的表达是否改变了 眼睛的免疫抑制性质。 最后，我们建议制作一个 第三种模型，通过在α-A的控制下表达I-E抗原 晶体蛋白启动子 在这些小鼠中，克隆缺失V β 17 a（I-E 通过单克隆抗体染色评估T细胞， 流式细胞仪 这些研究将提供重要的新信息， 对组织限制性抗原的耐受诱导机制，以及 为自身免疫性葡萄膜炎提供新的小鼠模型。