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、粘附和信号传导中起作用。在人类中，Wiskott - Aldrich综合征（WAS）是一种严重的x连锁免疫缺陷，由WASp的功能突变丧失引起，导致严重的免疫缺陷。相反，WAS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WASp的情况下，2型反应的升高可能是由先天免疫细胞而不是由经典的CD4+ Th2细胞驱动的。了解特定的效应缺陷在离散的白细胞亚群将是关键的病理适当治疗WAS。我们分离了CD4+ T细胞的WASp缺陷，并在体内和体外检测了WASp在CD4+ T细胞效应功能中的作用。使用这种方法，我们对2型免疫进行了关键观察，使我们对WAS中的免疫有了新的认识，并突出了WAS-/-先天和适应性区室的差异。我们假设，在人类WAS（高IgE和特应性皮炎）中，增强的2型反应与强大的先天IL-4和增强的Th2细胞向炎症组织的募集/保留相结合。特异性目标1：先天室中WASp的缺失是否驱动2型反应？我们已经建立了实验系统，将WASp缺陷限制在先天或适应性CD4细胞区室。我们将与Raif Geha博士合作，利用特应性皮炎（AD）模型，确定淋巴结（LN）和皮肤部位的2型先天和CD4+ T细胞的动力学、大小和效应功能。这一目标的数据将为WAS中Th2病理的主要贡献细胞类型提供第一个直接测试。特异性目标2：炎症组织中Th2募集/保留的增加是否补偿功能能力的降低？我们发现，与WT型Th2细胞相比，缺乏WASp的Th2细胞更容易在发炎的真皮中积聚，这表明WASp调节了组织的募集或保留。我们将在AD模型中使用FACS和全身成像（Xenogen IVIS）直接测试Th2和非t型2细胞的募集情况。对于组织内的保留，我们将使用活体双光子显微镜来评估间质运动性和T-APC接触时间，并使用过继转移到真皮部位来跟踪组织保留/退出。

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.