VISUALIZATION OF ANTIGEN-SPECIFIC T-CELLS IN ACAID

描述（申请人的描述）：眼睛中的免疫应答具有 适用于消除生物威胁， 炎症操纵这种高度调节的免疫反应可能导致 延长移植物植入的治疗策略，包括视网膜 细胞移植和减轻自身免疫性疾病。的注入 小鼠眼前房中的鸡卵清蛋白（OVA）诱导了 一种独特的全身耐受形式，称为前房相关免疫 偏差或ACAID。ACAID是研究免疫调节的极好模型。 在随后皮下注射免疫原性形式的OVA后，这些 动物表现出对延迟型超敏反应的引发减少。在 此外，我们已经表明，细胞溶解性T细胞（CTL）反应也减少， 在ACAID。这表明，CTL前体被删除或未能扩增， 很好。或者，CTL前体被扩增但功能失活。 为了确定CTL前体的命运，有必要跟踪这些细胞 in vivo.然而，抗原特异性CTL前体在幼稚以及 免疫的小鼠低于流式细胞术的检测水平。克服 这种障碍，T细胞，来自转基因小鼠，表达T细胞受体， 特异于与H-2 Kb复合的OVA肽257-264的蛋白质将被转移到 同基因小鼠。这种方法增加了CTL前体频率， 维持正常的生理免疫反应。供体T细胞将 通过流式细胞术在ACAID诱导后的受体小鼠中计数 使用0 VA 257 -264/H-2Kb四聚体分析。ACAID的生成包括 主动抑制脾脏CD 8 + T细胞已被证明是调节因子 传出抑制的可能性此外，我们已经证明， T细胞也有助于传出抑制。使用CTL的体外测定 抑制，我们将确定γ δ T细胞是否是传出的 如果γ δ T细胞诱导CD 8 + α T细胞， 成为传出抑制器。

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.