VISUALIZATION OF ANTIGEN-SPECIFIC T-CELLS IN ACAID

描述(申请人的描述)：眼睛中的免疫反应 适应于消除生物威胁，而不具有显著的非特异性 发炎。对这种高度调控的免疫反应的操纵可能会导致 延长移植植入期的治疗策略，包括视网膜 细胞移植和减轻自身免疫性疾病。注射…… 小鼠眼前房内鸡卵清蛋白(OVA)诱导的小鼠眼部炎症反应 一种独特的全身性耐受，称为前房相关免疫 偏差或ACAID。ACAID是研究免疫调节的良好模型。 在随后的皮下注射免疫原型OVA后，这些 动物表现出对迟发性超敏反应的启动减少。在……里面 此外，我们还发现，细胞溶解T细胞(CTL)的反应也降低了 在ACAID。这表明CTL前体被删除或无法在 ACAID。或者，CTL前体被扩展，但在功能上不被激活。 为了确定CTL前体的命运，有必要追踪这些细胞 在活体内。然而，抗原特异性CTL前体在幼稚和 免疫小鼠低于流式细胞仪检测水平。要克服 这种障碍，来自转基因小鼠的T细胞，表达T细胞受体 与H-2kb络合的OVA多肽257-264将被转移到 幼稚的同基因小鼠。这种方法提高了CTL前体的频率，同时 维持正常的生理免疫反应。捐献的T细胞将是 用流式细胞术检测ACAID诱导后受体小鼠的细胞计数 用OVA257-264/H-2kb四聚体分析。ACAID的产生包括 主动抑制。脾CD8+T细胞已被证明是调节器 ACAID的传出抑制。此外，我们已经证明了Gammadelta T细胞也有助于传出抑制。用体外CTL法检测CTL 抑制，我们将确定Gammadelta T细胞是否为传出细胞 ACAID抑制子或Gammadelta T细胞是否诱导CD8+Alphabeta T细胞 成为传出的抑制者。

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.