Mol Juliana P S, Costa Erica A, Carvalho Alex F, Sun Yao-Hui, Tsolis Reneé M, Paixão Tatiane A, Santos Renato L
Departamento de Clínica e Cirurgia Veterinárias, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Departamento de Medicina Veterinária Preventiva, Escola de Veterinária, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
PLoS One. 2014 Sep 26;9(9):e108606. doi: 10.1371/journal.pone.0108606. eCollection 2014.
The pathogenesis of the Brucella-induced inflammatory response in the bovine placenta is not completely understood. In this study we evaluated the role of the B. abortus Type IV secretion system and the anti-inflammatory factor BtpB in early interactions with bovine placental tissues. Transcription profiles of chorioallantoic membrane (CAM) explants inoculated with wild type (strain 2308), ΔvirB2 or ΔbtpB Brucella abortus were compared by microarray analysis at 4 hours post infection. Transcripts with significant variation (>2 fold change; P<0.05) were functionally classified, and transcripts related to defense and inflammation were assessed by quantitative real time RT-PCR. Infection with wild type B. abortus resulted in slightly more genes with decreased than increased transcription levels. Conversely, infection of trophoblastic cells with the ΔvirB2 or the ΔbtpB mutant strains, that lack a functional T4SS or that has impaired inhibition of TLR signaling, respectively, induced more upregulated than downregulated genes. Wild type Brucella abortus impaired transcription of host genes related to immune response when compared to ΔvirB and ΔbtpB mutants. Our findings suggest that proinflammatory genes are negatively modulated in bovine trophoblastic cells at early stages of infection. The virB operon and btpB are directly or indirectly related to modulation of these host genes. These results shed light on the early interactions between B. abortus and placental tissue that ultimately culminate in inflammatory pathology and abortion.
布鲁氏菌引起的牛胎盘炎症反应的发病机制尚未完全明确。在本研究中,我们评估了流产布鲁氏菌IV型分泌系统及抗炎因子BtpB在与牛胎盘组织早期相互作用中的作用。在感染后4小时,通过微阵列分析比较接种野生型(2308菌株)、ΔvirB2或ΔbtpB流产布鲁氏菌的绒膜尿囊膜(CAM)外植体的转录谱。对差异显著(>2倍变化;P<0.05)的转录本进行功能分类,并通过定量实时RT-PCR评估与防御和炎症相关的转录本。野生型流产布鲁氏菌感染导致转录水平降低的基因略多于转录水平升高的基因。相反,分别缺乏功能性IV型分泌系统或TLR信号抑制受损的ΔvirB2或ΔbtpB突变株感染滋养层细胞后,诱导上调的基因多于下调的基因。与ΔvirB和ΔbtpB突变株相比,野生型流产布鲁氏菌损害了宿主与免疫反应相关基因的转录。我们的研究结果表明,在感染早期,牛滋养层细胞中的促炎基因受到负调控。virB操纵子和btpB与这些宿主基因的调控直接或间接相关。这些结果揭示了流产布鲁氏菌与胎盘组织之间的早期相互作用,这些相互作用最终导致炎症病理和流产。