Oggioni Marco R, Trappetti Claudia, Kadioglu Aras, Cassone Marco, Iannelli Francesco, Ricci Susanna, Andrew Peter W, Pozzi Gianni
Laboratorio di Microbiologia Molecolare e Biotecnologia, Dipartimento di Biologia Molecolare, Università di Siena, Siena, Italy.
Mol Microbiol. 2006 Sep;61(5):1196-210. doi: 10.1111/j.1365-2958.2006.05310.x.
Two main patterns of gene expression of Streptococcus pneumoniae were observed during infection in the host by quantitative real time RT-PCR; one was characteristic of bacteria in blood and one of bacteria in tissue, such as brain and lung. Gene expression in blood was characterized by increased expression of pneumolysin, pspA and hrcA, while pneumococci in tissue infection showed increased expression of neuraminidases, metalloproteinases, oxidative stress and competence genes. In vitro situations with similar expression patterns were detected in liquid culture and in a newly developed pneumococcal model of biofilm respectively. The biofilm model was dependent on addition of synthetic competence stimulating peptide (CSP) and no biofilm was formed by CSP receptor mutants. As one of the differentially expressed gene sets in vivo were the competence genes, we exploited competence-specific tools to intervene on pneumococcal virulence during infection. Induction of the competence system by the quorum-sensing peptide, CSP, not only induced biofilm formation in vitro, but also increased virulence in pneumonia in vivo. In contrast, a mutant for the ComD receptor, which did not form biofilm, also showed reduced virulence in pneumonia. These results were opposite to those found in a bacteraemic sepsis model of infection, where the competence system was downregulated. When pneumococci in the different physiological states were used directly for challenge, sessile cells grown in a biofilm were more effective in inducing meningitis and pneumonia, while planktonic cells from liquid culture were more effective in inducing sepsis. Our data enable us, using in vivo gene expression and in vivo modulation of virulence, to postulate the distinction - from the pneumococcal point of view - between two main types of disease. During bacteraemic sepsis pneumococci resemble planktonic growth, while during tissue infection, such as pneumonia or meningitis, pneumococci are in a biofilm-like state.
通过定量实时逆转录聚合酶链反应(RT-PCR)观察到,肺炎链球菌在宿主体内感染期间存在两种主要的基因表达模式;一种是血液中细菌的特征性模式,另一种是组织(如脑和肺)中细菌的特征性模式。血液中的基因表达特征是肺炎溶血素、肺炎球菌表面蛋白A(PspA)和热休克调控蛋白A(HrcA)的表达增加,而组织感染中的肺炎球菌则表现出神经氨酸酶、金属蛋白酶、氧化应激和感受态基因的表达增加。在液体培养和新建立的肺炎球菌生物膜模型中分别检测到了具有相似表达模式的体外情况。生物膜模型依赖于添加合成的感受态刺激肽(CSP),CSP受体突变体不会形成生物膜。由于体内差异表达的基因集中有感受态基因,我们利用感受态特异性工具在感染期间干预肺炎球菌的毒力。群体感应肽CSP诱导感受态系统,不仅能在体外诱导生物膜形成,还能在体内增加肺炎的毒力。相比之下,不形成生物膜的ComD受体突变体在肺炎中的毒力也降低。这些结果与在菌血症败血症感染模型中发现的结果相反,在该模型中感受态系统被下调。当直接使用处于不同生理状态的肺炎球菌进行攻击时,生物膜中生长的固着细胞在诱导脑膜炎和肺炎方面更有效,而液体培养中的浮游细胞在诱导败血症方面更有效。我们的数据使我们能够利用体内基因表达和体内毒力调节,从肺炎球菌的角度推测两种主要疾病类型之间的区别。在菌血症败血症期间,肺炎球菌类似于浮游生长,而在组织感染(如肺炎或脑膜炎)期间,肺炎球菌处于生物膜样状态。