State Key Laboratory of Veterinary Biotechnology and Zoonosis Laboratory of the Ministry of Agriculture, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang, People's Republic of China.
PLoS One. 2011;6(12):e28485. doi: 10.1371/journal.pone.0028485. Epub 2011 Dec 28.
Brucella melitensis is an important zoonotic pathogen that causes brucellosis, a disease that affects sheep, cattle and occasionally humans. B. melitensis strain M5-90, a live attenuated vaccine cultured from B. melitensis strain M28, has been used as an effective tool in the control of brucellosis in goats and sheep in China. However, the molecular changes leading to attenuated virulence and pathogenicity in B. melitensis remain poorly understood. In this study we employed the Illumina Genome Analyzer platform to perform genome-wide digital gene expression (DGE) analysis of mouse peritoneal macrophage responses to B. melitensis infection. Many parallel changes in gene expression profiles were observed in M28- and M5-90-infected macrophages, suggesting that they employ similar survival strategies, notably the induction of anti-inflammatory and antiapoptotic factors. Moreover, 1019 differentially expressed macrophage transcripts were identified 4 h after infection with the different B. melitensis strains, and these differential transcripts notably identified genes involved in the lysosome and mitogen-activated protein kinase (MAPK) pathways. Further analysis employed gene ontology (GO) analysis: high-enrichment GOs identified endocytosis, inflammatory, apoptosis, and transport pathways. Path-Net and Signal-Net analysis highlighted the MAPK pathway as the key regulatory pathway. Moreover, the key differentially expressed genes of the significant pathways were apoptosis-related. These findings demonstrate previously unrecognized changes in gene transcription that are associated with B. melitensis infection of macrophages, and the central signaling pathways identified here merit further investigation. Our data provide new insights into the molecular attenuation mechanism of strain M5-90 and will facilitate the generation of new attenuated vaccine strains with enhanced efficacy.
马耳他布鲁氏菌是一种重要的人畜共患病病原体,可引起布鲁氏菌病,该病影响绵羊、牛,偶尔也会影响人类。M5-90 株是由 M28 株培养而来的活减毒疫苗,已被用作中国控制山羊和绵羊布鲁氏菌病的有效工具。然而,马耳他布鲁氏菌减毒和毒力丧失的分子变化仍知之甚少。在这项研究中,我们使用 Illumina 基因组分析仪平台对小鼠腹腔巨噬细胞对布鲁氏菌感染的全基因组数字基因表达(DGE)分析。在 M28 和 M5-90 感染的巨噬细胞中观察到许多基因表达谱的平行变化,这表明它们采用了相似的生存策略,特别是诱导抗炎和抗凋亡因子。此外,在感染不同的布鲁氏菌菌株 4 小时后,鉴定出 1019 个差异表达的巨噬细胞转录本,这些差异转录本特别鉴定出与溶酶体和丝裂原激活蛋白激酶(MAPK)途径相关的基因。进一步的分析采用基因本体论(GO)分析:高富集 GO 鉴定出内吞作用、炎症、凋亡和运输途径。Path-Net 和 Signal-Net 分析强调了 MAPK 途径是关键的调控途径。此外,显著途径的关键差异表达基因与凋亡有关。这些发现表明与巨噬细胞感染布鲁氏菌相关的基因转录发生了先前未被识别的变化,并且这里确定的关键信号通路值得进一步研究。我们的数据为 M5-90 株的分子减毒机制提供了新的见解,并将有助于生成具有增强功效的新型减毒疫苗株。