Disciplina de Microbiologia, Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, Brazil.
Department of Microbiology, UT Southwestern Medical Center, Dallas, Texas, USA.
Microbiol Spectr. 2024 Oct 3;12(10):e0030424. doi: 10.1128/spectrum.00304-24. Epub 2024 Aug 27.
Atypical enteropathogenic (aEPEC) is a significant cause of diarrhea in low- and middle-income countries. Certain aEPEC strains, including the Brazilian representative strain of serotype O51:H40 called aEPEC 1711-4, can use flagella to attach to, invade, and persist in T84 and Caco-2 intestinal cells. It can also translocate from the gut to extraintestinal sites in a rat model. Although various aspects of the virulence of this strain were studied and the requirement of a type III secretion system for the efficiency of the invasion process was demonstrated, the expression of the locus of enterocyte effacement (LEE) genes during the invasion and intracellular persistence remains unclear. To address this question, the expression of flagella and the different LEE operons was evaluated during kinetic experiments of the interaction of aEPEC 1711-4 with enterocytes . The genome of the strain was also sequenced. The results showed that flagella expression remained unchanged, but the expression of and increased during the early interaction and invasion of aEPEC 1711-4 into Caco-2 cells, and there was no change 24 h post-infection during the persistence period. The number of actin accumulation foci formed on HeLa cells also increased during the 6-h analysis. No known gene related to the invasion process was identified in the genome of aEPEC 1711-4, which was shown to belong to the global EPEC lineage 10. These findings suggest that the LEE components and the intimate adherence promoted by intimin are necessary for the invasion and persistence of aEPEC 1711-4, but the detailed mechanism needs further study.IMPORTANCEAtypical enteropathogenic (aEPEC) is a major cause of diarrhea, especially in low- and middle-income countries, like Brazil. However, due to the genome heterogeneity of each clonal group, it is difficult to comprehend the pathogenicity of this strain fully. Among aEPEC strains, 1711-4 can invade eukaryotic cells , cross the gut barrier, and reach extraintestinal sites in animal models. By studying how different known aEPEC virulence factors are expressed during the invasion process, we can gain insight into the commonalities of this phenotype among other aEPEC strains. This will help in developing preventive measures to control infections caused by invasive strains. No known virulence-encoding genes linked to the invasion process were found. Nevertheless, additional studies are still necessary to evaluate the role of other factors in this phenotype.
非典型肠致病性大肠杆菌(aEPEC)是中低收入国家腹泻的重要病因。某些 aEPEC 菌株,包括巴西 O51:H40 血清型代表菌株 aEPEC 1711-4,可利用鞭毛附着、入侵并在 T84 和 Caco-2 肠细胞中持续存在。它还可以在大鼠模型中从肠道转移到肠外部位。尽管对该菌株的各种毒力方面进行了研究,并证明了 III 型分泌系统对入侵过程效率的要求,但在入侵和细胞内持续存在期间,肠上皮细胞脱落(LEE)基因座的表达仍不清楚。为了解决这个问题,在 aEPEC 1711-4 与肠细胞相互作用的动力学实验中,评估了鞭毛和不同 LEE 操纵子的表达。还对该菌株的基因组进行了测序。结果表明,在 aEPEC 1711-4 早期与 Caco-2 细胞相互作用和入侵过程中,鞭毛表达保持不变,但 和 的表达增加,在感染后 24 小时的持续期内没有变化。在 6 小时的分析中,在 HeLa 细胞上形成的肌动蛋白聚集焦点的数量也增加了。在 aEPEC 1711-4 的基因组中未发现与入侵过程相关的已知基因,该基因被证明属于全球 EPEC 谱系 10。这些发现表明,LEE 成分和紧密附着促进的紧密素对于 aEPEC 1711-4 的入侵和持续存在是必要的,但详细的机制需要进一步研究。
重要性 非典型肠致病性大肠杆菌(aEPEC)是腹泻的主要病因,特别是在巴西等中低收入国家。然而,由于每个克隆群的基因组异质性,很难全面理解该菌株的致病性。在 aEPEC 菌株中,1711-4 可入侵真核细胞 ,穿过肠道屏障,并在动物模型中到达肠外部位。通过研究不同已知的 aEPEC 毒力因子在入侵过程中的表达情况,可以深入了解其他 aEPEC 菌株中这种表型的共性。这将有助于制定预防措施来控制由侵袭性菌株引起的感染。在入侵过程中没有发现与毒力相关的编码基因。然而,仍需要进一步研究来评估其他因素在这种表型中的作用。