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菌毛编码基因yadC对多种生物学特性具有多效性作用,并在禽致病性大肠杆菌致病性中发挥作用。

Fimbria-Encoding Gene yadC Has a Pleiotropic Effect on Several Biological Characteristics and Plays a Role in Avian Pathogenic Escherichia coli Pathogenicity.

作者信息

Verma Renu, Rojas Thaís Cabrera Galvão, Maluta Renato Pariz, Leite Janaína Luisa, da Silva Livia Pilatti Mendes, Nakazato Gerson, Dias da Silveira Wanderley

机构信息

Department of Genetics, Evolution, and Bioagents, Institute of Biology, University of Campinas-UNICAMP, Campinas, SP, Brazil

Department of Genetics, Evolution, and Bioagents, Institute of Biology, University of Campinas-UNICAMP, Campinas, SP, Brazil.

出版信息

Infect Immun. 2015 Oct 26;84(1):187-93. doi: 10.1128/IAI.01138-15. Print 2016 Jan.

Abstract

The extraintestinal pathogen termed avian pathogenic Escherichia coli (APEC) is known to cause colibacillosis in chickens. The molecular basis of APEC pathogenesis is not fully elucidated yet. In this work, we deleted a component of the Yad gene cluster (yadC) in order to understand the role of Yad in the pathogenicity of the APEC strain SCI-07. In vitro, the transcription level of yadC was upregulated at 41°C and downregulated at 22°C. The yadC expression in vivo was more pronounced in lungs than in spleen, suggesting a role in the early steps of the infection. Chicks infected with the wild-type and mutant strains presented, respectively, 80% and 50% mortality rates. The ΔyadC strain presented a slightly decreased ability to adhere to HeLa cells with or without the d-mannose analog compared with the wild type. Real-time PCR (RT-PCR) assays showed that fimH was downregulated (P < 0.05) and csgA and ecpA were slightly upregulated in the mutant strain, showing that yadC modulates expression of other fimbriae. Bacterial internalization studies showed that the ΔyadC strain had a lower number of intracellular bacteria recovered from Hep-2 cells and HD11 cells than the wild-type strain (P < 0.05). Motility assays in soft agar demonstrated that the ΔyadC strain was less motile than the wild type (P < 0.01). Curiously, flagellum-associated genes were not dramatically downregulated in the ΔyadC strain. Taken together, the results show that the fimbrial adhesin Yad contributes to the pathogenicity and modulates different biological characteristics of the APEC strain SCI-07.

摘要

被称为禽致病性大肠杆菌(APEC)的肠道外病原体已知会在鸡中引起大肠杆菌病。APEC发病机制的分子基础尚未完全阐明。在这项研究中,我们删除了Yad基因簇的一个组成部分(yadC),以了解Yad在APEC菌株SCI-07致病性中的作用。在体外,yadC的转录水平在41°C时上调,在22°C时下调。yadC在体内的表达在肺中比在脾中更明显,表明其在感染早期阶段发挥作用。用野生型和突变株感染的雏鸡死亡率分别为80%和50%。与野生型相比,ΔyadC菌株在有或没有d-甘露糖类似物的情况下粘附HeLa细胞的能力略有下降。实时PCR(RT-PCR)分析表明,突变株中fimH下调(P<0.05),csgA和ecpA略有上调,表明yadC调节其他菌毛的表达。细菌内化研究表明,从Hep-2细胞和HD11细胞中回收的细胞内细菌数量,ΔyadC菌株比野生型菌株少(P<0.05)。软琼脂中的运动性分析表明,ΔyadC菌株的运动性比野生型菌株弱(P<0.01)。奇怪的是,鞭毛相关基因在ΔyadC菌株中没有显著下调。综上所述,结果表明菌毛粘附素Yad有助于APEC菌株SCI-07的致病性并调节其不同的生物学特性。

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