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通过基因表达和新生小鼠死亡率评估水性和气单胞菌临床分离株的毒力,随后基于转录反应评估细胞培养预测毒力的能力。

Evaluating virulence of waterborne and clinical Aeromonas isolates using gene expression and mortality in neonatal mice followed by assessing cell culture's ability to predict virulence based on transcriptional response.

作者信息

Hayes S L, Rodgers M R, Lye D J, Stelma G N, McKinstry C A, Malard J M, Vesper S J

机构信息

USEPA, National Risk Management Research Laboratory, Water Supply/Water Resources Division, Cincinnati, OH 45268, USA.

出版信息

J Appl Microbiol. 2007 Oct;103(4):811-20. doi: 10.1111/j.1365-2672.2007.03318.x.

DOI:10.1111/j.1365-2672.2007.03318.x
PMID:17897183
Abstract

AIMS

To assess the virulence of Aeromonas spp. using two models, a neonatal mouse assay and a mouse intestinal cell culture.

METHODS AND RESULTS

After artificial infection with a variety of Aeromonas spp., mRNA extracts from the two models were processed and hydridized to murine microarrays to determine host gene response. Definition of virulence was determined based on host mRNA production in murine neonatal intestinal tissue and mortality of infected animals. Infections of mouse intestinal cell cultures were then performed to determine whether this simpler model system's mRNA responses correlated to neonatal results and therefore be predictive of virulence of Aeromonas spp. Virulent aeromonads up-regulated transcripts in both models including multiple host defense gene products (chemokines, regulation of transcription and apoptosis and cell signalling). Avirulent species exhibited little or no host response in neonates. Mortality results correlated well with both bacterial dose and average fold change of up-regulated transcripts in the neonatal mice.

CONCLUSIONS

Cell culture results were less discriminating but showed promise as potentially being able to be predictive of virulence. Jun oncogene up-regulation in murine cell culture is potentially predictive of Aeromonas virulence.

SIGNIFICANCE AND IMPACT OF THE STUDY

Having the ability to determine virulence of waterborne pathogens quickly would potentially assist public health officials to rapidly assess exposure risks.

摘要

目的

使用两种模型,即新生小鼠试验和小鼠肠道细胞培养,评估气单胞菌属的毒力。

方法与结果

用多种气单胞菌属进行人工感染后,对两种模型的mRNA提取物进行处理,并与小鼠微阵列杂交以确定宿主基因反应。根据小鼠新生肠道组织中的宿主mRNA产生情况和受感染动物的死亡率来确定毒力的定义。然后进行小鼠肠道细胞培养感染,以确定这个更简单的模型系统的mRNA反应是否与新生小鼠的结果相关,从而能否预测气单胞菌属的毒力。在两种模型中,有毒力的气单胞菌上调了转录本,包括多种宿主防御基因产物(趋化因子、转录调控、细胞凋亡和细胞信号传导)。无毒力的菌株在新生小鼠中几乎没有或没有宿主反应。死亡率结果与细菌剂量和新生小鼠中转录本上调的平均倍数变化都有很好的相关性。

结论

细胞培养结果的区分度较低,但有望能够预测毒力。小鼠细胞培养中Jun癌基因的上调可能预示气单胞菌的毒力。

研究的意义和影响

能够快速确定水生病原体的毒力可能会帮助公共卫生官员迅速评估接触风险。

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