VISUALIZATION OF ANTIGEN-SPECIFIC T-CELLS IN ACAID

ACAID 中抗原特异性 T 细胞的可视化

• 批准号: 6298922
• 负责人: KYLE C MCKENNA
• 金额: $ 3.24万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6298922
• 关键词: RNase protection assay; T cell receptor; T lymphocyte; anterior chamber; antigen antibody reaction; cellular immunity; cytokine; cytotoxic T lymphocyte; delayed hypersensitivity; flow cytometry; genetically modified animals; imaging /visualization /scanning; immune tolerance /unresponsiveness; laboratory mouse; suppressor T lymphocyte; tumor antigens

DESCRIPTION (Applicant's Description): The immune response in the eye has adapted to eliminate biological threats without significant nonspecific inflammation. Manipulation of this highly regulated immune response may lead to therapeutic strategies for prolonging transplant engraftment, including retinal cell transplantation, and mitigating autoimmune diseases. The injection of chicken ovalbumin (OVA) in the anterior chamber of the eye of mice induces a unique form of systemic tolerance called anterior chamber associated immune deviation or ACAID. ACAID is an excellent model for studying immune regulation. Upon a subsequent subcutaneous injection with an immunogenic form of OVA, these animals display reduced priming for delayed-type hypersensitivity responses. In addition, we have shown that cytolytic T-cell (CTL) responses are also reduced in ACAID. This suggests that CTL precursors are deleted or fail to expand in ACAID. Alternatively, CTL precursors are expanded but functionally inactivated. To determine the fate of CTL precursors, it is necessary to track these cells in vivo. However, antigen-specific CTL precursors in naive as well as immunized mice are below the level of detection by flow cytometry. To overcome this obstacle, T-cells, from transgenic mice, expressing a T-cell receptor specific to OVA peptide 257-264 complexed with H-2 Kb, will be transferred into naive syngeneic mice. This method increases the CTL precursor frequency while maintaining a normal physiological immune response. Donor T-cells will be enumerated in recipient mice following ACAID induction by flow cytometric analysis using OVA257-264/H-2Kb tetramers. The generation of ACAID includes active suppression. Splenic CD8+ T-cells have been shown to be the regulators of efferent suppression in ACAID. In addition, we have shown that gammadelta T-cells also contribute to efferent suppression. Using an in vitro assay of CTL suppression, we will determine if gammadelta T-cells are the efferent suppressor in ACAID or if gammadelta T-cells induce CD8+ alphabeta T-cells to become the efferent suppressor.

描述（申请人的描述）：眼睛中的免疫反应 适应消除无明显非特异性的生物威胁 炎。操纵这种高度调节的免疫反应可能导致 延长移植植入的治疗策略，包括视网膜 细胞移植和减轻自身免疫性疾病。注射 小鼠眼前腔室的鸡卵形蛋白（OVA）诱导A 独特形式的全身耐受性称为前室相关免疫 偏差或ACAID。 ACAID是研究免疫调节的绝佳模型。 在随后的皮下注射中，具有免疫原性的OVA，这些 动物表现出减少延迟型超敏反应的启动。在 此外，我们已经表明了细胞溶解T细胞（CTL）的反应也会降低 在Acaid中。这表明CTL前体被删除或无法扩展 Acaid。或者，CTL前体被扩展，但在功能上灭活。 为了确定CTL前体的命运，有必要跟踪这些细胞 体内。但是，幼稚和抗原特异性的CTL前体 免疫小鼠通过流式细胞仪低于检测水平。克服 来自转基因小鼠的这种障碍物T细胞表达T细胞受体 特定于与H-2 Kb复合的OVA肽257-264将转移到 幼稚的同步小鼠。此方法增加了CTL前体频率，而 保持正常的生理免疫反应。捐赠者T细胞将是 通过流式细胞术诱导Acaid诱导后的接受者小鼠列举 使用OVA257-264/H-2KB四聚体进行分析。 Acaid的一代包括 主动抑制。脾脏CD8+ T细胞已被证明是调节剂 Acaid中的传出抑制。此外，我们已经表明了伽马得舞 T细胞也有助于传出抑制。使用CTL的体外测定 抑制作用，我们将确定gammadelta t细胞是否是传出 Acaid中的抑制剂或gammadelta t细胞诱导CD8+ Alphabeta T细胞 成为传出抑制器。