Innate vs Adaptive Contributions to Th2 Pathology in Wiskott Aldrich Syndrome

描述(由申请人提供)：Wiskott-Aldrich综合征蛋白(WASP)是所有造血细胞中肌动蛋白聚合的关键调节因子，在白细胞迁移、黏附和信号传递中发挥作用。在人类中，Wiskott-Aldrich综合征(Was)是一种严重的X连锁免疫缺陷，原因是黄蜂功能突变导致免疫严重缺陷。相反，患者表现出IgE水平升高和特应性皮炎/湿疹。我们在小鼠中的数据表明，在没有黄蜂的情况下，2型反应的升高可能是由先天免疫细胞驱动的，而不是由经典的CD4+Th2细胞驱动的。了解离散白细胞亚群中的特定效应器缺陷对于适当治疗风湿性心脏病的病理至关重要。我们分离了黄蜂对CD4+T细胞的缺陷，并在体内外检测了黄蜂对CD4+T细胞效应器功能的影响。使用这种方法，我们对2型免疫进行了关键的观察，为我们提供了一个新的免疫处理方法，并强调了先天免疫和适应性免疫之间的差异。我们假设，人类的2型反应增强(高IgE和特应性皮炎)与强大的固有IL-4和增强的Th2细胞对炎症组织的募集/保留有关。具体目标1：先天隔间中黄蜂的丢失是否会驱动II型反应？我们已经产生了实验系统，将黄蜂缺乏限制在先天或适应性的CD4隔间。我们将与Raif Geha博士合作，使用特应性皮炎(AD)模型来确定淋巴结(LN)和真皮部位的2型固有T细胞和CD4+T细胞的动力学、大小和效应功能。来自这一目的的数据将提供第一个直接测试主要贡献细胞类型(S)的Th2型细胞病理。具体目标2：炎症组织中Th2募集/滞留的增加是否弥补了功能能力的下降？我们发现缺乏黄蜂的Th2细胞比WT Th2细胞更容易在发炎的真皮中聚集，这表明黄蜂调节组织招募或滞留。我们将在AD模型中用流式细胞仪和全身成像(Xenogen IVIS)直接检测Th2和非T2型细胞的募集。对于组织内的滞留，我们将使用活体双光子显微镜来评估组织间的运动性和T-APC接触时间，并使用过继转移到真皮部位来跟踪组织滞留/退出。 公共卫生相关性：到目前为止，Wiskott-Aldrich综合征(Was)患者的免疫缺陷是通过分析个体免疫细胞类型的功能缺陷来表征的。然而，目前还不清楚这些个体的异常是如何导致Wiskott-Aldrich综合征的病理的。我们建议使用黄蜂缺乏的小鼠模型来确定不同的免疫区段在人类观察到的2型病理的发展中所起的作用：高IgE和特应性皮炎。这些知识对于设计细胞特异性疗法以恢复AS患者的有效免疫力是必不可少的。

DESCRIPTION (provided by applicant): The Wiskott-Aldrich Syndrome protein (WASp) is a key regulator of actin polymerization in all hematopoietic cells and functions in leukocyte migration, adhesion and signaling. In humans, the Wiskott - Aldrich syndrome (WAS) is a severe X-linked immunodeficiency caused by loss of function mutations in WASp that lead to profound defects in immunity. In contrast, WAS patients exhibit increased IgE levels and atopic dermatitis/eczema. Our data in the mouse suggest that the elevated Type 2 response in the absence of WASp may be driven by innate immune cells rather than by classic CD4+ Th2 cells. Understanding specific effector defects in discrete leukocyte subsets will be critical for the appropriate treatment of pathology in WAS. We isolated the WASp deficiency to CD4+ T cells and examined the role of WASp in CD4+ T cell effector function in vitro and in vivo. Using this approach we have made key observations on Type 2 immunity that give us a novel handle on immunity in WAS and highlight differences in WAS-/- innate and adaptive compartments. We hypothesize that the heightened Type 2 response in human WAS (high IgE and atopic dermatitis) is compounded by robust innate IL-4 and enhanced recruitment/retention of Th2 cells to inflamed tissue. Specific Aim 1: Does loss of WASp in the innate compartment drive the Type 2 response? We have generated experimental systems that limit WASp deficiency to either innate or adaptive CD4 compartment. We will determine the kinetics, magnitude and effector function of Type 2 innate and CD4+ T cells at lymph node (LN) and dermal sites using an atopic dermatitis (AD) model in collaboration with Dr Raif Geha. Data from this aim will provide the first direct test of the principal contributing cell type(s) to Th2 pathology in WAS. Specific Aim 2: Does increased Th2 recruitment/retention in inflamed tissue compensate for reduced functional capacity? We find that WASp-deficient Th2 cells accumulate in the inflamed dermis more readily than WT Th2 cells suggesting WASp regulates tissue recruitment or retention. We will directly test recruitment of Th2 and non-T Type-2 cells with FACS and whole body imaging (Xenogen IVIS) in the AD model. For retention within the tissue we will use intravital 2-photon microscopy to assess interstitial motility and T-APC contact time and use adoptive transfer into the dermal site to follow tissue retention/exit. PUBLIC HEALTH RELEVANCE: The immune deficiencies in Wiskott-Aldrich Syndrome (WAS) patients have been characterized thus far by analysis of defects in the function of individual immune cell types. It is unclear however exactly how these individual abnormalities contribute to the pathologies of the Wiskott-Aldrich Syndrome. We propose to use the mouse model of WASp deficiency to determine the contributions of distinct immune compartments on the development of the Type 2 pathology observed in human WERE: high IgE and atopic dermatitis. Such knowledge will be essential for the design of cell-specific therapies that seek to restore effective immunity in WAS patients.