Urinary exosome as a potential biomarker for urinary tract infection
Urinary exosome as a potential biomarker for urinary tract infection
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DOI：
10.1111/cmi.13020
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发表时间：
2019-07-01
影响因子：
3.4
通讯作者：
Ito, Masafumi
中科院分区：
文献类型：
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作者：
Mizutani, Kosuke;Kawakami, Kyojiro;Ito, Masafumi
与尿路感染(UTI)不同，无症状性细菌尿(ABU)不应接受治疗，但孕妇和将接受创伤性泌尿外科干预的患者除外。然而，目前还没有临床可用的标记物来鉴别诊断。胞外体或小的胞外小泡携带细胞中含有的蛋白质，因此有可能成为几种疾病的生物标志物。基于尿外切体在尿路感染和ABU患者中的分子特征可能不同的假设，我们检验了尿外切体是否可以作为鉴别诊断的标志。用超速离心法或亲和层析法从人尿液和单核细胞培养上清液中分离出外切体。用Western印迹分析、ELISA法和CLEIA法检测蛋白质的表达。结果表明，在THP-1和SV-Huc-1细胞与大肠杆菌共培养和/或内毒素处理后，细胞内信号分子Akt和ERK以及转录因子NF-kappa B的水平增加。在尿路感染患者的尿外切体中，Akt显著降低，外切体标志物CD9经抗菌药物治疗后有下降趋势。更重要的是，尿路感染患者尿Exosome中Akt和CD9水平高于ABU患者，经尿肌酐校正后也可观察到这一点。总之，这些结果提示尿Exosome中Akt和CD9可作为鉴别诊断UTI和ABU的有用标记物。
Unlike urinary tract infection (UTI), asymptomatic bacteriuria (ABU) should not be treated, with some exceptions such as pregnant women and patients who will undergo traumatic urologic interventions. However, there has been no clinically available marker for their differential diagnosis. Exosomes or small extracellular vesicles carry proteins contained in cells from which they are derived, thus having the potential as a biomarker of several diseases. On the basis of the hypothesis that the molecular signature of exosomes in urine may differ between UTI and ABU patients, we examined if urinary exosomes could serve as a marker for their differential diagnosis. Exosomes were isolated by ultracentrifugation or affinity-based method from cell culture medium of monocytic THP-1 and uroepithelial SV-HUC-1 cells and human urine. Protein expression was examined by Western blot analysis, ELISA, and CLEIA. The results showed that the levels of intracellular signalling molecules Akt and ERK and transcription factor NF-kappa B increased in exosomes isolated from THP-1 and SV-HUC-1 cells cocultured with Escherichia coli and/or treated with lipopolysaccharide. In urinary exosomes of UTI patients, Akt significantly diminished, and an exosomal marker CD9 showed a trend to decrease after treatment with antimicrobial agents. More importantly, Akt and CD9 levels in urinary exosomes were higher in UTI patients than in ABU patients, which was also observed after correction by urine creatinine. Collectively, these results suggest that Akt and CD9 in urinary exosomes could be useful markers for differential diagnosis of UTI and ABU.