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对自然感染牛分枝杆菌的欧洲野猪下颌淋巴结和口咽扁桃体中差异应激/炎症反应的蛋白质组学和转录组学分析。

Proteomic and transcriptomic analyses of differential stress/inflammatory responses in mandibular lymph nodes and oropharyngeal tonsils of European wild boars naturally infected with Mycobacterium bovis.

作者信息

Naranjo Victoria, Villar Margarita, Martín-Hernando Maria Paz, Vidal Dolors, Höfle Ursula, Gortazar Christian, Kocan Katherine M, Vázquez Jesús, de la Fuente José

机构信息

Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), Ciudad Real, Spain.

出版信息

Proteomics. 2007 Jan;7(2):220-31. doi: 10.1002/pmic.200600527.

Abstract

Differential stress/inflammatory responses were characterized at the mRNA and protein levels in mandibular lymph nodes (MLN) and oropharyngeal tonsils of European wild boars (Sus scrofa), naturally infected with Mycobacterium bovis. Suppression-subtractive hybridization combined with immunohistochemistry and/or quantitative real-time RT-PCR were used to identify and characterize abundant stress/inflammatory gene sequences differentially expressed in tuberculous (TB+) wild boars. Genes identified in MLN and tonsils corresponded to serum amyloid A, arginase I, osteopontin, lysozyme, annexin I, and heat shock proteins, respectively. Global protein patterns in MLN and tonsils were compared between TB+ and nontuberculous (TB-) boars by 2-DE and MALDI-TOF MS. Five proteins, including stress/inflammatory proteins annexin V, serum albumin, and apolipoprotein A1 were found at lower levels in MLN of TB+ boars. Manganese superoxide dismutase was found up-regulated in MLN of TB+ boars. Five proteins, including creatine kinase and MHC class II antigens were found up-regulated in tonsils of TB+ boars. These results demonstrated differential stress/inflammatory responses in wild boars naturally infected with M. bovis and suggest possible markers of tuberculosis in this species that may prove useful for future studies of host-pathogen interactions and for diagnostics and vaccine development.

摘要

对自然感染牛分枝杆菌的欧洲野猪(Sus scrofa)的下颌淋巴结(MLN)和口咽扁桃体中的差异应激/炎症反应进行了mRNA和蛋白质水平的表征。采用抑制性消减杂交结合免疫组织化学和/或定量实时RT-PCR来鉴定和表征在结核性(TB+)野猪中差异表达的丰富应激/炎症基因序列。在MLN和扁桃体中鉴定出的基因分别对应于血清淀粉样蛋白A、精氨酸酶I、骨桥蛋白、溶菌酶、膜联蛋白I和热休克蛋白。通过二维电泳(2-DE)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)比较了TB+和非结核性(TB-)野猪的MLN和扁桃体中的整体蛋白质模式。在TB+野猪的MLN中发现包括应激/炎症蛋白膜联蛋白V、血清白蛋白和载脂蛋白A1在内的5种蛋白质水平较低。在TB+野猪的MLN中发现锰超氧化物歧化酶上调。在TB+野猪的扁桃体中发现包括肌酸激酶和MHC II类抗原在内的5种蛋白质上调。这些结果证明了自然感染牛分枝杆菌的野猪存在差异应激/炎症反应,并提示该物种中可能的结核病标志物,这可能对未来宿主-病原体相互作用的研究以及诊断和疫苗开发有用。

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