Naranjo Victoria, Höfle Ursula, Vicente Joaquín, Martín M Paz, Ruiz-Fons Francisco, Gortazar Christian, Kocan Katherine M, de la Fuente José
Instituto de Investigación en Recursos Cinegéticos, Ronda de Toledo, Ciudad Real, Spain.
FEMS Immunol Med Microbiol. 2006 Mar;46(2):298-312. doi: 10.1111/j.1574-695X.2005.00035.x.
Bovine tuberculosis, caused by Mycobacterium bovis (Mycobacterium tuberculosis complex), is a zoonotic disease that affects cattle worldwide. The bacterium infects other animal species, both domesticated and wild, and this range of hosts complicates attempts to control or eradicate the disease. Despite advances in the characterization of the mechanisms involved in host-pathogen interactions and host cell responses to M. tuberculosis complex in human, bovine and mouse cells, differentially expressed genes in tissue biopsies of naturally occurring tuberculous and nontuberculous exposed individuals have been poorly characterized. In this study, differential gene expression was analysed using suppression-subtractive hybridization in oropharyngeal tonsils and mandibular lymph nodes of field-collected tuberculous and nontuberculous European wild boars from a tuberculosis-endemic area of Spain. Real-time PCR and semiquantitative reverse-transcriptase PCR of selected genes confirmed the results of the suppression-subtractive hybridization analysis. Protein expression of selected differentially expressed genes was analysed by radial immunodiffusion or immunohistochemistry. Differential gene expression varied among tuberculous and non-tuberculous groups and between tonsils and lymph nodes. Single and multiple cellular mechanisms were affected, including signal transduction, immune response, inflammation, stress, apoptosis/antiapoptosis, cell structure, adhesion and transport, protein and DNA/RNA metabolism and enzymatic processes. These results demonstrate the modulation of gene expression by mycobacterial infection in tonsils and mandibular lymph nodes of European wild boars naturally exposed to M. bovis, and provide a basis for defining host-pathogen interactions and the mechanism of protective immunity.
牛结核病由牛分枝杆菌(结核分枝杆菌复合群)引起,是一种影响全球牛群的人畜共患病。该细菌可感染其他家养和野生动物物种,宿主范围广泛使得控制或根除该疾病的努力变得复杂。尽管在人类、牛和小鼠细胞中宿主-病原体相互作用以及宿主细胞对结核分枝杆菌复合群反应的机制研究方面取得了进展,但对自然感染结核和未感染结核个体组织活检中差异表达基因的特征了解甚少。在本研究中,利用抑制性消减杂交技术分析了从西班牙结核病流行地区野外采集的患结核病和未患结核病的欧洲野猪的口咽扁桃体和下颌淋巴结中的差异基因表达。对选定基因进行实时PCR和半定量逆转录PCR,证实了抑制性消减杂交分析的结果。通过放射免疫扩散或免疫组织化学分析选定差异表达基因的蛋白质表达。结核组和非结核组之间以及扁桃体和淋巴结之间的差异基因表达各不相同。单细胞和多细胞机制均受到影响,包括信号转导、免疫反应、炎症、应激、凋亡/抗凋亡、细胞结构、黏附与转运、蛋白质和DNA/RNA代谢以及酶促过程。这些结果证明了自然暴露于牛分枝杆菌的欧洲野猪扁桃体和下颌淋巴结中分枝杆菌感染对基因表达的调节作用,并为确定宿主-病原体相互作用及保护性免疫机制提供了依据。