2012-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/683984摘要：過敏的盛行率在已開發國家中仍然穩定地增加，像是一些過敏性鼻炎、結膜炎、皮膚炎或是氣喘的病人，他們的病症都是由IgE 來調控。當敏感性體質的人吸入黴菌的孢子後，會產生過敏的症狀，因此黴菌被視為重要過敏原之一。在台灣最常見的室內黴菌為Penicillium sp.以及Aspergillus sp.，其中Penicillium sp.的培養液萃取物中，含有許多可與IgE 抗體高度結合的過敏原。一些具有蛋白酶活性的過敏原，已經被鑑定為一群主要的過敏原，並且能引發上皮細胞脫屑及前發炎細胞激素的釋放。Pen c 13 為這些過敏原的其中一員，它是由Penicillium citrinum 所分泌，分子量為33-kD 的主要過敏原，先前一些活體外的實驗也發現，由於許多的絲胺酸蛋白水解酶都可以降解細胞間的接合蛋白，因此能夠改變上皮細胞的通透性，最近也有一些研究證明蛋白水解酶在促進過敏的敏化作用以及過敏性呼吸道發炎中扮演重要角色。在此，我們建立一個老鼠模式，經由支氣管注射的方式，連續每天給予Pen c 13，可導致老鼠血清的total IgE，Pen c 13-specific IgE 和IgG1 顯著上升，與Th1 相關的Penc 13-specific IgG2a 則無差異性。此外我們亦發現到，Pen c 13 能夠讓NCI-H441 細胞株的TEER 值下降並且亦能切割黏著連接蛋白: E-cadherin 和緊密連接蛋白: occludin, ZO-1等，這些結果意味著透過環境中的蛋白水解酶來打開細胞間的連接，是氣喘疾病發展的第一步。本計畫中，我們將建立一個最佳的條件來産生過敏性肺部疾病之實驗動物模式，經由連續吸入Pen c 13 而沒有佐劑的情況，同時也將結合蛋白質體學與生物資訊學方法，找出加速肺部微環境敏化作用的可能的機制。<br> Abstract: The prevalence of allergy has been increasing steadily in the developed world.IgE-mediated allergy is a serious problem for patients with symptoms such as allergic rhinitis,conjunctivitis, dermatitis or asthma. It has long been recognized that inhalation of fungalspores can produce allergic symptoms in susceptible individuals and fungi are regarded as oneof the main sources of allergens. Penicillium and Aspergillus are the most common indoorfungal species in Taiwan. In Penicillium spp., many allergens show higher IgE-bindingactivity in culture filtrate extracts. Some allergens with protease activity have been identifiedas a group of major allergens and cause epithelial cell desquamation and release ofproinflammatory cytokines. One of allergens, Pen c 13, is the 33-kD immunodominantallergen secreted by Penicillium citrinum. Prevously studies demonstrate that many serineproteases induce permeability of epithelial cells in vitro due to their ability to degrade thejunctional proteins. A number of recent studies have also identified a critical role for proteasesin promoting allergic sensitization and allergic airway inflammation.Here we established a mouse model by exposing to active Pen c 13 (intratracheally) daily,resulted in a significant increase serum levels of total IgE, Pen c 13-specific IgE and IgG1.However, no significant differences were observed in Th1-associated Pen c 13-specific IgG2abetween any of the experimental groups. Based on our recently in vitro studies, in confluentNCI-H441 cell which is human lung adenocarcinoma epithelial cell line, Pen c 13 lead to thedecrease of TEER value and the cleavage of adherent junctional protein: E-cadherin and tightjunctional proteins: occludin, ZO-1. These results suggest that opening of intercellular junctionsby environmental protease may be the initial step in the development of asthma. In this project,we will set up an optimal condition to develop experimental animal models of allergic lungdiseases by prolonged inhalation of Pen c 13 without adjuvant. Furthermore, we will alsocombine proteomics and bioinformatics approaches to identify the possible mechanisms tocontribute a lung microenvironment that fosters the development of allergic sensitization.