首页> 美国卫生研究院文献>Journal of Bacteriology >The Oxidoreductase DsbA Plays a Key Role in the Ability of the Crohns Disease-Associated Adherent-Invasive Escherichia coli Strain LF82 To Resist Macrophage Killing
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The Oxidoreductase DsbA Plays a Key Role in the Ability of the Crohns Disease-Associated Adherent-Invasive Escherichia coli Strain LF82 To Resist Macrophage Killing

机译:氧化还原酶DsbA在克罗恩氏病相关的黏附侵袭性大肠杆菌菌株LF82抵抗巨噬细胞杀伤能力中发挥关键作用

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摘要

Adherent-invasive Escherichia coli (AIEC) isolated from Crohn's disease patients is able to adhere to and invade intestinal epithelial cells and to replicate in mature phagolysosomes within macrophages. Here, we show that the dsbA gene, encoding a periplasmic oxidoreductase, was required for AIEC strain LF82 to adhere to intestinal epithelial cells and to survive within macrophages. The LF82-ΔdsbA mutant did not express flagella and, probably as a consequence of this, did not express type 1 pili. The role of DsbA in adhesion is restricted to the loss of flagella and type 1 pili, as forced contact between bacteria and cells and induced expression of type 1 pili restored the wild-type phenotype. In contrast, the dsbA gene is essential for AIEC LF82 bacteria to survive within macrophages, irrespective of the loss of flagella and type 1 pilus expression, and the survival ability of LF82-ΔdsbA was as low as that of the nonpathogenic E. coli K-12, which was efficiently killed by macrophages. We also provide evidence that the dsbA gene is needed for LF82 bacteria to grow and survive in an acidic and nutrient-poor medium that partly mimics the harsh environment of the phagocytic vacuole. In addition, under such stress conditions dsbA transcription is highly up-regulated. Finally, the CpxRA signaling pathway does not play a role in regulation of dsbA expression in AIEC LF82 bacteria under conditions similar to those of mature phagolysosomes.
机译:从克罗恩氏病患者中分离出的粘附侵袭性大肠杆菌(AIEC)能够粘附并侵入肠道上皮细胞,并在巨噬细胞内的成熟吞噬体中复制。在这里,我们显示dsbA基因,编码周质氧化还原酶,是AIEC菌株LF82粘附于肠上皮细胞并在巨噬细胞中生存所必需的。 LF82-ΔdsbA突变体不表达鞭毛,并且可能因此而不表达1型菌毛。 DsbA在粘附中的作用仅限于鞭毛和1型菌毛的丧失,因为细菌和细胞之间的强制接触以及1型菌毛的诱导表达恢复了野生型表型。相反,dsbA基因对于AIEC LF82细菌在巨噬细胞中生存至关重要,而不论鞭毛损失和1型菌毛表达如何,LF82-ΔdsbA的生存能力都与非致病性大肠杆菌K-一样低。 12,它被巨噬细胞有效杀死。我们还提供了证据,表明dsbA基因是LF82细菌在酸性和营养贫乏的培养基中生长和生存所需的,该培养基部分模拟了吞噬液泡的恶劣环境。此外,在这种压力条件下,dsbA转录高度上调。最后,在类似于成熟吞噬体的条件下,CpxRA信号通路在AIEC LF82细菌中的dsbA表达调节中不起作用。

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