Bacterial Mast cell conditioning modulates skin allergic reactions

 描述（由申请人提供）：我们的研究结果表明，肥大细胞（MC）在皮肤中可能具有不同的细胞因子谱，并且皮肤微生物组对塑造这些不同的谱具有强烈的影响。我们已经证明，皮肤表面的细菌产物在人类MC中诱导抗炎表型，这有助于维持皮肤稳态并有助于避免不必要的炎症。在这项提案中，我们的目标是证明由皮肤微生物组产品调节的MC可以在与人类免疫系统相关的背景下塑造皮肤对过敏原的反应。使用在体内和体外试验，我们打算定义的相对贡献的MC-细菌的相互作用，在指导树突状细胞和T细胞的贩运，以应对过敏原。在初步研究中，我们 使用来自两种代表性细菌菌株的组分：病原体S.金黄色葡萄球菌群体获得了耐甲氧西林的USA 300（USA 300）和耐甲氧西林金黄色葡萄球菌。表皮细胞，并表明它们都能够改变MC炎性表型。我们将使用这些细菌成分来探索在过敏反应期间操纵MC的可能性，最终目标是开发用于治疗过敏性炎症和相关皮肤疾病的新治疗策略。为了表征皮肤中MC-细菌相互作用的影响，我们提出以下内容：1.测试用细菌产品预处理MC是否导致体内皮肤过敏反应的调节。我们假设，暴露于MCs的肠道细菌小分子将诱导抗炎MC细胞因子的配置文件，导致皮肤免疫环境的变化和皮肤过敏反应的减少。本研究采用两种不同的小鼠皮肤变态反应模型，探讨MC预处理对小鼠皮肤变态反应的影响。我们还将在离体系统中使用激光捕获显微切割（LCM）验证LTA和细菌产物对人皮肤中MC IL-10表达的影响。为此目的，体内过敏反应研究产生的数据将阐明MC产生的细胞因子谱的重要性以及它们如何调节皮肤的过敏反应。2.目的探讨细菌制品预处理MCs对树突状细胞细胞因子表达的影响及其调节过敏反应的机制。我们假设，MCs预处理与海藻酸副产物或LTA单独，将防止或减少树突状细胞（DC）分化和T细胞成熟在体外，从而抑制过敏反应。在这个建议的第二部分，我们将把我们的兴趣集中在APC细胞因子，这是负责T细胞极化。这项体外研究产生的数据将为我们提供关于预处理的MC释放细胞因子如何扭曲过敏反应的机制的重要信息。

 DESCRIPTION (provided by applicant): Our findings suggest that Mast cells (MCs) can have different cytokine profiles in the skin and that the skin microbiome has a strong influence on shaping these different profiles. We have demonstrated that bacterial products at the skin surface induce an anti-inflammatory phenotype in human MCs that contributes to maintaining skin homeostasis and helps avoid unnecessary inflammation. In this proposal, we aim to demonstrate that MCs, conditioned by the skin microbiome products, can shape skin responses to allergens in contexts that are relevant to the human immune system. Using both in vivo and in vitro assays, we intend to define the relative contribution of MC-bacterial interactions in directing dendritic and T cell trafficking in response to allergens. In our preliminary studies, we employ components from two representative bacterial strains: the pathogen S. aureus community acquired methicillin resistant USA 300 (USA 300) and the commensal S. epidermidis and show that they are both capable of shifting MC inflammatory phenotypes. We will use these bacterial components to explore the possibility of manipulating MCs during allergic reactions with the end goal of developing new therapeutic strategies for the treatment of allergic inflammation and related skin diseases. To characterize of the effects of MC-bacterial interactions in the skin, we propose the followings: 1. To test whether preconditioning of MCs with bacterial products results in the modulation of skin allergic reactions in vivo. We hypothesize that exposure of MCs to commensal bacterial small molecules will induce anti-inflammatory MC cytokine profiles, resulting in changes to the skin immunological environment and the reduction of the allergic response in the skin. Using two different mouse models of allergic reaction we will investigate the effects of MC preconditioning on allergic reactions in th skin. We will also verify the effects of LTA and bacterial products on MC IL-10 expression in human skin using laser capture micro dissection (LCM) in an ex-vivo system. Data generated by the in vivo allergic reaction studies in this aim will clarify the importance of the cytokine profies generated by MCs and how they can modulate allergic reactions in the skin. 2. To investigate the mechanism of how preconditioning of MCs with bacterial products modifies dendritic cell cytokine profiles to modulate allergic reactions in vitro. We hypothesize that MCs preconditioned with commensal byproducts or LTA alone, will prevent or decrease dendritic cell (DC) differentiation and T cell maturation in vitro and consequently dampen the allergic response. In the second part of this proposal, we will focus our interest on APC cytokines, which are responsible for T cell polarization. Data generated by this in vitro study will give us important information regarding the mechanism of how the release of cytokines by preconditioned MCs can skew the allergic response.