REGULATION OF ACIAD BY LYMPHOCYTES WITH NK MARKERS

几十年来，免疫学家已经知道免疫是细胞介导的和抗体介导的反应的各种贡献的结果。近年来，可能的免疫应答的二分法已经通过以下知识被进一步定义：T辅助细胞可以采用两种极性类型的反应性之一：T辅助细胞1（Th1）和T辅助细胞2（Th2）。这些对抗反应可以通过它们产生的细胞因子（Th1、IFN γ; Th2、IL 4、IL 10）的表型和抗体同种型（Th1：IgG 2a; Th2：IgG 1、IgE）的差异来区分。Th1应答与炎症相关，而Th2应答与炎症相关，而Th2应答与超敏反应和抵抗寄生虫入侵相关。由于这两种极性反应相互交叉调节，因此Th2反应可能在预防包括眼睛在内的各种器官的炎症中很重要。显然，在健康眼中，由于前房中不寻常的局部调节机制，很难诱导Th1应答。该研究提案将研究称为前房相关免疫偏离（ACAID）的眼部调节机制。ACAID与Th2应答的上调和Th1应答的下调有关，即炎症，从而保护视力。在动物模型中，将置于前房（a.c.）眼睛引起的系统性免疫反应，这是明显不寻常的是有一个选择性缺乏迟发型超敏反应（DTH，一个指标的Th1，反应）。虽然这个模型已经得到了很好的研究，但还没有人研究过可能被调用来产生和维持眼部Th2应答的早期机制。我们建议，两个不同的淋巴细胞亚群表达自然杀伤（NK）抗原可能参与接种到前房的抗原的极性反应的早期调节。我们实验室的最新数据表明，ACID不会在CD1缺陷小鼠中发展。本申请将通过研究NKT细胞和CD 1的机制和相互作用来利用这一观察结果。我们将使用多种方法来验证假设，包括流式细胞术与细胞内细胞因子和表面标志物的同时标记，组织和悬浮液中细胞的形态学分析，分子生物学技术，包括RNA酶保护试验。此外，我们将评估NK抗原阳性淋巴细胞在整个动物模型中的作用，该模型被操作以预防ACID，并比较幼稚和致敏小鼠中的ACAID。这些研究的结果将为理解眼睛免疫偏离的细胞机制提供新的信息，并对NKT（先天免疫）和适应性免疫反应的相互作用产生新的见解。

Immunologists have known for decades that immunity is the result of the various contributions of cell-mediated and antibody-mediated responses. Within recent years the dichotomy of possible immune responses has been further defined by the knowledge that T helper cells can adopt one of two polar types of reactivity: T helper 1 (Th1) and T helper 2 (Th2). These counter responses can be differentiated by differences in the phenotype of cytokines (Th1, IFNgamma; Th2, IL4, IL10) and isotypes of antibodies (Th1: IgG2a; Th2: IgG1, IgE) they make. Th1 responses are associated with inflammation, while Th2 responses are associated with inflammation, while Th2 responses associated with hypersensitivity and protection against parasitic invasion. Because these two polar responses cross regulate each other it is possible that Th2 responses might be important in the prevention of inflammation in various organs including the eye. Apparently, in the health eye, it is very difficult to induce a Th1 response because of an unusual local regulatory mechanism in the anterior chamber. This research proposal will study the ocular regulatory mechanism known as Anterior Chamber Associated Immune Deviation (ACAID). ACAID is relevant to the up-regulation of Th2 responses and the down regulation of Th1 responses, i.e. inflammation, thereby preserving sight. In an animal model antigenic materials that are placed in the anterior chamber (a.c.) Of the eye evoke a systemic immune response that is distinctly unusual in there is a selective deficiency of delayed-type hypersensitivity (DTH, an indicator of a Th1, response). While this model is well studied, no one has looked at early mechanisms that might be invoked to produce and maintain an ocular Th2 response. We propose that two distinct subpopulations of lymphocytes expressing natural killer (NK) antigens may participate in the early regulation of the polar responses to antigens inoculated into the anterior chamber. Recent data from our laboratory show that ACID does not develop in CD1 deficient mice. This application will exploit this observation by studying the mechanisms and interaction of NKT cells and CD1. We will use a variety of methods to test the hypotheses including flow cytometry with simultaneous labeling of intracellular cytokines and surface markers, morphological analysis of cells in tissues and suspension, molecular biology techniques, including Rnase protection assay. In addition we will evaluate the role of NK antigen positive lymphocytes in whole animal models that are manipulated to prevent ACID and compare ACAID in naive and primed mice. Resents of the studies will generate new information toward understanding the cellular mechanisms of immune deviation in the eye and new insights into the interactions of NKT (innate immunity) and the adaptive immune response.