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，由WASp功能缺失突变引起，导致免疫力严重缺陷。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，在缺乏WASp的情况下，升高的2型应答可能是由先天免疫细胞而不是经典的CD 4 + Th 2细胞驱动的。了解离散白细胞亚群中的特定效应缺陷对于WAS病理学的适当治疗至关重要。我们分离了WASp缺陷的CD 4 + T细胞，并在体外和体内研究了WASp在CD 4 + 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，这为我们提供了一种新的WAS免疫处理方法，并突出了WAS-/-先天和适应性区室的差异。我们假设，人类WAS（高IgE和特应性皮炎）中2型反应的增强与强大的先天IL-4和Th 2细胞向炎症组织的募集/保留增强有关。具体目标1：先天性室中WASp的丢失是否驱动2型反应？我们已经产生了实验系统，限制WASp缺陷先天或适应性CD 4区室。我们将与Raif Geha博士合作，使用特应性皮炎（AD）模型确定淋巴结（LN）和真皮部位2型先天性和CD 4 + T细胞的动力学，幅度和效应功能。来自该目标的数据将提供对WAS中Th 2病理学的主要贡献细胞类型的首次直接测试。具体目标2：炎症组织中增加的Th 2募集/保留是否补偿了功能能力的降低？我们发现WASp缺陷型Th 2细胞比WT Th 2细胞更容易在发炎的真皮中积累，这表明WASp调节组织募集或保留。我们将在AD模型中用FACS和全身成像（Xenogen IVIS）直接测试Th 2和非T 2型细胞的募集。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以跟踪组织保留/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.