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从肾结石患者尿液中分离的大肠杆菌中的延伸因子 Tu 可促进草酸钙晶体生长和聚集。

Elongation factor Tu on Escherichia coli isolated from urine of kidney stone patients promotes calcium oxalate crystal growth and aggregation.

机构信息

Faculty of Graduate School, Khon Kaen University, Khon Kaen, Thailand.

Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand.

出版信息

Sci Rep. 2017 Jun 7;7(1):2953. doi: 10.1038/s41598-017-03213-x.

Abstract

Escherichia coli is the most common bacterium isolated from urine and stone matrix of calcium oxalate (CaOx) stone formers. Whether it has pathogenic role(s) in kidney stone formation or is only entrapped inside the stone remains unclear. We thus evaluated differences between E. coli isolated from urine of patients with kidney stone (EUK) and that from patients with urinary tract infection (UTI) without stone (EUU). From 100 stone formers and 200 UTI patients, only four pairs of EUK/EUU isolates had identical antimicrobial susceptibility patterns. Proteomic analysis revealed nine common differentially expressed proteins. Among these, the greater level of elongation factor Tu (EF-Tu) in EUK was validated by Western blotting. Outer membrane vesicles (OMVs) derived from EUK had greater promoting activities on CaOx crystallization, crystal growth and aggregation as compared to those derived from EUU. Neutralizing the OMVs of EUK with monoclonal anti-EF-Tu antibody, not with an isotype antibody, significantly reduced all these OMVs-induced promoting effects. Moreover, immunofluorescence staining of EF-Tu on bacterial cell surface confirmed the greater expression of surface EF-Tu on EUK (vs. EUU). Our data indicate that surface EF-Tu and OMVs play significant roles in promoting activities of E. coli on CaOx crystallization, crystal growth and aggregation.

摘要

大肠埃希菌是从尿液和草酸钙(CaOx)结石形成者的结石基质中分离出的最常见细菌。它在肾结石形成中是否具有致病作用,还是仅被包裹在结石内,目前尚不清楚。因此,我们评估了从肾结石患者尿液中分离出的大肠杆菌(EUK)和从无结石尿路感染患者尿液中分离出的大肠杆菌(EUU)之间的差异。从 100 名结石患者和 200 名尿路感染患者中,只有 4 对 EUK/EUU 分离株具有相同的抗菌药物敏感性模式。蛋白质组学分析显示了 9 种常见的差异表达蛋白。其中,EUK 中的延伸因子 Tu(EF-Tu)水平较高,通过 Western blot 得到验证。与 EUU 衍生的 OMVs 相比,EUK 衍生的 OMVs 对 CaOx 结晶、晶体生长和聚集具有更大的促进作用。用单克隆抗 EF-Tu 抗体(而不是同种型抗体)中和 EUK 的 OMVs,可显著降低所有这些 OMVs 诱导的促进作用。此外,EF-Tu 在细菌细胞表面的免疫荧光染色证实了 EUK 上表面 EF-Tu 的表达高于 EUU(与 EUU 相比)。我们的数据表明,表面 EF-Tu 和 OMVs 在促进大肠杆菌对 CaOx 结晶、晶体生长和聚集的活性中起重要作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/359e/5462744/5917f5578955/41598_2017_3213_Fig1_HTML.jpg

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