Xu Xiao-Yong, Chen Fei, Sun He, Chen Chen, Zhao Bei-Lei
Department of Respiratory Medicine, Nanjing General Hospital of Nanjing Military Command, PLA Nanjing, Jiangsu, China.
Am J Transl Res. 2016 May 15;8(5):2419-25. eCollection 2016.
Aspergillus is widely distributed in the Earth's biosphere. It has strong adaptive capacity, and lives as saprophytic or parasitic life. This study aims to investigate the role of E-cadherin for adhesion of Aspergillus fumigatus blastospores in a human epithelial cell line (A549) and search the correlated molecule in aspergillus. A. fumigatus blastospores were incubated with the total protein of A549 to investigate the binding of E-cadherin and blastospores followed by an affinity purification procedure. After establishing the adhesion model, the adhesion of A. fumigatus blastospores by A549 cells was evaluated by down-regulating E-cadherin of A549 cells with small interfering RNA (siRNA). FVB mice constructed with E-cadherin down-regulation were infected with aspergillus fumigatus. Preliminary exploration of E-cadherin interacting protein on the surface of aspergillus fumigates by immunoprecipitation and mass spectrometry analysis. E-cadherin was adhered to the surface of A. fumigatus blastospore. Adhesion of the blastospores was reduced by blocking or down-regulating E-cadherin in A549 cells. E-cadherin showed limited significance in the process of mice against aspergillus fumigates. Mass spectrometry (MS) analysis indicated the following proteins AFUA_8G07080, AfA24A6.130c, XP_747789 can bind to E-cadherin. In conclusion, E-cadherin is a receptor for adhesion of A. fumigatus blastospores in epithelial cells. This may open a new approach to treat this fungal infection.
曲霉菌广泛分布于地球生物圈。它具有很强的适应能力,以腐生或寄生方式生存。本研究旨在探讨E-钙黏蛋白在烟曲霉芽生孢子黏附人上皮细胞系(A549)中的作用,并寻找曲霉菌中的相关分子。将烟曲霉芽生孢子与A549的总蛋白孵育,通过亲和纯化程序研究E-钙黏蛋白与芽生孢子的结合。建立黏附模型后,用小干扰RNA(siRNA)下调A549细胞的E-钙黏蛋白,评估A549细胞对烟曲霉芽生孢子的黏附。构建E-钙黏蛋白下调的FVB小鼠,用烟曲霉感染。通过免疫沉淀和质谱分析初步探索烟曲霉表面与E-钙黏蛋白相互作用的蛋白。E-钙黏蛋白黏附于烟曲霉芽生孢子表面。在A549细胞中阻断或下调E-钙黏蛋白可降低芽生孢子的黏附。E-钙黏蛋白在小鼠抗烟曲霉过程中意义有限。质谱(MS)分析表明,AFUA_8G07080、AfA24A6.130c、XP_747789等蛋白可与E-钙黏蛋白结合。总之,E-钙黏蛋白是上皮细胞中烟曲霉芽生孢子黏附的受体。这可能为治疗这种真菌感染开辟新途径。