Allergic inflammation in genetic models of lung ILC2 deficiency

描述(由申请人提供)：先天2型淋巴细胞(ILC2，也称为天然辅助细胞或核细胞)与广泛的传染病和过敏性疾病密切相关。通过停留在粘膜屏障并对上皮源性细胞因子如IL-33和IL-25做出反应，它们迅速释放早期的2型细胞因子，促进适应性CD4T细胞的Th2炎症反应和B细胞的IgE产生。然而，在缺乏分子签名的情况下，ILC2谱系的定义仍然是试探性的，并存在一定的争议。此外，尽管它们与其他适应性和先天淋巴细胞来自相同的共同淋巴前体(CLP)，但它们的发育中间产物及其与其他相关谱系的关系尚不清楚。这个项目建立在我们实验室对PLZF-IRES-GFPCre报告鼠的初步研究基础上，表明转录因子PLZF，以前被认为是先天类IL-4/IL-13产生NKT细胞谱系的标志，在ILC2的发育过程中也有高水平的表达。核心假设是PLZF在ILC2的发育过程中发挥关键作用，PLZF报告/缺失小鼠可以用来从遗传上跟踪和操纵ILC2前体及其成熟后代。本应用的目的是利用PLZF-IRES-GFPCre小鼠鉴定ILC2的骨髓前体细胞，鉴定它们的分子程序，并通过与rosa26-FL-Stop-FL-Tomato小鼠杂交后的命运定位来确定其后代。此外，该项目将测试在体内通过基因操作或消融ILC2的各种方法，作为表征它们在生理和过敏性呼吸道炎症中的功能的前奏。其具体目的是：1)鉴定CLP的PLZFhi亚群；2)确定PLZF在ILC2发育和功能中的作用；3)建立ILC2缺陷小鼠模型，用于过敏性呼吸道炎症的研究。该项目具有创新性，因为它探索了PLZF表达在专门研究2型反应的先天和先天样谱系中共享的新想法，还因为它将为体内ILC2的研究产生新的模型和试剂。这将标志着我们理解和进一步操纵先天免疫在由2型免疫引起或治愈的世界性疾病中的作用的能力发生了重大转变。这项拟议的研究具有重要意义，因为它将极大地增强我们对先天淋巴细胞谱系发育的理解。开发的概念和工具将使开发针对寄生虫和过敏原的不同天然免疫成分的试剂成为可能。

DESCRIPTION (provided by applicant): Innate type 2 lymphocytes (ILC2, also called natural helper cells or nuocytes) are critically involved in widespread infectious and allergic diseases. By residing at mucosal barriers and responding to epithelial-derived cytokines such as IL-33 and IL-25, they rapidly release the early type 2 cytokines that promote the Th2 inflammatory responses of adaptive CD4 T cells and the IgE production of B cells. However, the definition of the ILC2 lineage remains tentative and somewhat controversial in the absence of molecular signature. Furthermore, while they derive from the same common lymphoid precursors (CLP) as other adaptive and innate lymphocytes, their developmental intermediates and their relationship with other related lineages are unknown. This project builds on preliminary studies of PLZF-IRES-GFPCre reporter mice produced in our laboratory showing that the transcription factor PLZF, previously identified as the signature of the innate-like IL- 4/IL-13-producing NKT cell lineage, is also expressed at high levels during the development of ILC2s. The central hypothesis is that PLZF exerts critical functions in the development of ILC2s and that PLZF- reporter/deleter mice can be used to genetically track and manipulate ILC2 precursors and their mature progeny. The objective of this application is to use PLZF-IRES-GFPCre mice to identify the bone marrow precursors of ILC2, characterize their molecular program and define their progeny through fate mapping after crossing to ROSA26-FL-STOP-FL-Tomato mice. Furthermore, the project will test various approaches to genetically manipulate or ablate ILC2 in vivo as a prelude to characterizing their function in physiology and in allergic airway inflammation. The specific aims are: 1) to characterize the PLZFhi subset of CLP; 2) to determine the role of PLZF in ILC2 development and function; 3) to create ILC2-defective mouse models for studies of allergic airway inflammation. The project is innovative because it explores the novel idea that PLZF expression is shared across innate and innate-like lineages specializing in type 2 responses and because it will produce new models and reagents for studies of ILC2s in vivo. This would signal a major shift in our ability to understand and further manipulate the role of innate immunity in worldwide diseases caused or cured by type 2 immunity. The proposed research is significant because it will considerably enhance our understanding of innate lymphocyte lineage development. The concepts and tools developed will make it possible to develop agents that target distinct components of innate immunity to parasites and allergens.