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首页> 外文期刊>Infection and immunity >Identification of In Vivo-Induced Antigens Including an RTX Family Exoprotein Required for Uropathogenic Escherichia coli Virulence
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Identification of In Vivo-Induced Antigens Including an RTX Family Exoprotein Required for Uropathogenic Escherichia coli Virulence

机译：鉴定包括致病性大肠杆菌毒力所需的RTX家族外蛋白在内的体内诱导抗原

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Uncomplicated urinary tract infections (UTI) are caused most commonly by uropathogenic Escherichia coli (UPEC). Whole-genome screening approaches, including transcriptomic, proteomic, and signature-tagged mutagenesis, have shown that UPEC highly expresses or requires genes for translational machinery, capsule, lipopolysaccharide, type 1 fimbriae, and iron acquisition systems during UTI. To identify additional genes expressed by UPEC during UTI, an immunoscreening approach termed in vivo-induced antigen technology (IVIAT) was employed to identify antigens produced during experimental infection that are not produced during in vitro culture. An inducible protein expression library, constructed from genomic DNA isolated from UPEC strain CFT073, was screened using exhaustively adsorbed pooled sera from 20 chronically infected female CBA/J mice. Using this approach, we identified 93 antigens induced by UPEC in vivo. A representative subset of these genes was tested by quantitative PCR for expression by CFT073 in vivo and during growth in human urine or LB medium in vitro; proWX, narJI, lolA, lolD, tosA (upxA), c2432, katG, ydhX, kpsS, and yddQ were poorly expressed in vitro but highly expressed in vivo. Of these, tosA, a gene encoding a predicted repeat-in-toxin family member, was expressed exclusively during UTI. Deletion of tosA in UPEC strain CFT073 resulted in significant attenuation in bladder and kidney infections during ascending UTI. By screening for in vivo-induced antigens, we identified a novel UPEC virulence factor and additional proteins that could be useful as potential vaccine targets.
机译：尿路致病性大肠埃希菌（UPEC）最常引起单纯性尿路感染（UTI）。全基因组筛选方法，包括转录组学，蛋白质组学和标记标签的诱变，已经表明UPEC在UTI期间高表达或需要翻译机械，胶囊，脂多糖，1型菌毛和铁捕获系统的基因。为了识别UPEC在UTI期间表达的其他基因，采用了一种称为“体内诱导”抗原技术（IVIAT）的免疫筛选方法来鉴定实验感染过程中产生的抗原，这些抗原在体外< / em>文化。使用从20例慢性感染的雌性CBA / J小鼠中彻底吸收的合并血清，筛选由UPEC菌株CFT073分离的基因组DNA构建的诱导型蛋白表达文库。使用这种方法，我们鉴定了UPEC体内诱导的93种抗原。通过定量PCR测试了这些基因的代表性子集，以通过CFT073在体内以及在人尿或LB培养基中的生长过程中进行表达。 proWX ， narJI ， lolA ， lolD ， tosA （ upxA < / em>），c2432， katG ， ydhX ， kpsS 和 yddQ 在体外，但在体内高度表达。其中， tosA 是一种编码预测的毒素重复家族成员的基因，仅在UTI中表达。 UPEC菌株CFT073中 tosA 的缺失导致UTI升高期间膀胱和肾脏感染的明显减弱。通过筛选体内诱导的抗原，我们鉴定了一种新型的UPEC毒力因子和其他可用作潜在疫苗靶标的蛋白质。

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