TSG6 in IgE mediated Food Allergy

IgE 介导的食物过敏中的 TSG6

• 批准号: 7537680
• 负责人: Carine Blanchard
• 金额: $ 18.75万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-18 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7537680
• 关键词: Allergens; Anaphylaxis; Applications Grants; Attention; Attenuated; Binding; Biological Assay; Cell Adhesion; Clinical; Condition; Development; Diarrhea; Disease; End Point; Endopeptidases; Enzyme-Linked Immunosorbent Assay; Eosinophilic Esophagitis; Flow Cytometry; Food; Food Hypersensitivity; Gastrointestinal tract structure; Genes; Goals; Human; IgE; Immune response; Immunohistochemistry; In Situ Hybridization; Inflammation; Intestines; Kinetics; Laboratories; Life; Lymphoid Cell; Mediating; Messenger RNA; Modeling; Molecular; Mus; Neutrophil Infiltration; Oral; Pattern; Peptide Hydrolases; Polymerase Chain Reaction; Proteins; Role; System; Techniques; Time; Tissue Stains; Tissues; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; cell type; eosinophil; human TNF protein; in vivo; inhibitor/antagonist; novel therapeutics; protein expression; response

DESCRIPTION (provided by applicant): Food allergy and food anaphylaxis are debilitating and life threatening conditions. In order to better understand the molecular mechanisms underlying food anaphylaxis, our laboratory has extensively studied a murine model of oral allergen-induced IgE-dependent intestinal anaphylaxis (1). Interestingly, we recently uncovered that this model was characterized by an increased expression of the secreted protein TSG6 mRNA, tumor necrosis factor induced gene 6. At the same time, we also discovered that TSG6 mRNA was upregulated in a human food allergy related disease, eosinophilic esophagitis (EE) (2). Taken together, these sets of observations have focused our attention on uncovering the role of TSG6 in food allergy related responses. TSG6 is a potent inhibitor of neutrophil recruitment; some studies have indirectly associated TSG6 to lymphoid cell adhesion and activation through its propensity to increase binding to hyaluroan (6-8); and TSG6 increases protease activity. Our present study will focus on understanding the expression and role of TSG6 in food allergy-related responses in mice and humans. Aim I: To characterize TSG6 expression in murine and human food allergy related responses. In this aim, we hypothesize that TSG6 mRNA and protein levels will be upregulated during induction of experimental intestinal anaphylaxis, experimental EE, and human EE. We will define the kinetics and cell types that express TSG6 in both systems. Real-time PCR and ELISA will be used to quantify the mRNA and protein expression in mice. Immunohistochemistry, in situ hybridization, and flow cytometry analysis will be carried out to identify the TSG6 expression pattern in the gastrointestinal tract. Aim II: To define the role of TSG6 in vivo in experimental intestinal anaphylaxis. In this aim, we hypothesize that TSG6-deficient mice will develop attenuated Th2-associated diseases, i.e., a delay in diarrhea occurrence. Accordingly, we will assess a variety of clinical, pathological, and immunological endpoints using ELISA, flow cytometry, and tissue staining techniques. Aim III: To define the role of TSG6 in experimental EE. In this aim, we hypothesize that TSG6 will promote the development of EE, especially in eosinophil recruitment and tissue remodeling while inhibiting neutrophilic inflammation. TSG6-deficient mice will be subjected to experimental EE and the development of adaptive and innate immune responses will be assessed, as well as tissue remodeling using ELISA, immunohistochemistry, and protease activity assays. Potential impact: We are hopeful that this study will unravel new molecular mechanisms involved in IgE- mediated inflammation and will help provide a better understanding of the disease. We believe that this study will be a milestone in the development of new therapeutic approaches aiming to target TSG6 for IgE-mediated food allergy related diseases. Food allergy and food anaphylaxis are debilitating and life threatening conditions. The overall goal of these grant application is to uncover the role of TSG6 in food allergy related responses.

描述（由申请人提供）：食物过敏和食物过敏反应是使人衰弱和危及生命的疾病。为了更好地了解食物过敏反应的分子机制，我们的实验室广泛研究了口服过敏原诱导的IgE依赖性肠道过敏反应的小鼠模型（1）。有趣的是，我们最近发现，该模型的特征在于分泌蛋白TSG 6 mRNA，肿瘤坏死因子诱导基因6的表达增加。同时，我们还发现TSG 6 mRNA在人类食物过敏相关疾病嗜酸性粒细胞性食管炎（EE）中上调（2）。总之，这些观察结果使我们的注意力集中在揭示TSG 6在食物过敏相关反应中的作用上。TSG 6是一种有效的中性粒细胞募集抑制剂;一些研究通过其增加与透明质酸结合的倾向将TSG 6与淋巴细胞粘附和活化间接相关（6-8）; TSG 6增加蛋白酶活性。我们目前的研究将集中在了解TSG 6在小鼠和人类食物过敏相关反应中的表达和作用。目的I：表征TSG 6在小鼠和人食物过敏相关反应中的表达。在这个目标中，我们假设TSG 6的mRNA和蛋白水平将上调诱导实验性肠道过敏反应，实验EE，和人类EE. We将定义的动力学和细胞类型，在这两个系统中表达TSG 6。将使用实时PCR和ELISA来定量小鼠中的mRNA和蛋白质表达。将进行免疫组织化学、原位杂交和流式细胞术分析，以确定胃肠道中的TSG 6表达模式。目的二：明确TSG 6在实验性肠道过敏反应中的作用。为此，我们假设TSG 6缺陷小鼠将发展减弱的Th 2相关疾病，即，延迟腹泻的发生。因此，我们将使用ELISA、流式细胞术和组织染色技术评估各种临床、病理学和免疫学终点。目标三：为了确定TSG 6在实验性EE中的作用。为此，我们假设TSG 6将促进EE的发展，特别是在嗜酸性粒细胞募集和组织重塑方面，同时抑制中性粒细胞炎症。将对TSG 6缺陷型小鼠进行实验性EE，并使用ELISA、免疫组织化学和蛋白酶活性测定评估适应性和先天性免疫应答的发展以及组织重塑。潜在影响：我们希望这项研究将揭示IgE介导的炎症的新分子机制，并有助于更好地了解这种疾病。我们相信，这项研究将是一个里程碑，在发展新的治疗方法，旨在靶向TSG 6的IgE介导的食物过敏相关疾病。食物过敏和食物过敏反应是使人衰弱和危及生命的疾病。这些资助申请的总体目标是揭示TSG 6在食物过敏相关反应中的作用。