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Involvement of Quorum Sensing and Heat-Stable Enterotoxin a in Cell Damage Caused by a Porcine Enterotoxigenic Escherichia coli Strain

Zhu, Jing, Yin, Xianhua, Yu, Hai, Zhao, Liping, Sabour, Parviz, Gong, Joshua
Infection and immunity 2011 v.79 no.4 pp. 1688-1695
antibiotics, cell death, death, diarrhea, enterotoxigenic Escherichia coli, fimbriae, gene overexpression, genes, heat stability, neonates, pathogenesis, pathogens, piglets, quorum sensing
Enterotoxigenic Escherichia coli (ETEC) strains with K88 fimbriae are often associated with the outbreaks of diarrhea in newborn and weaned piglets worldwide. In the present study, we observed that 10⁸ CFU/ml of K88⁺ ETEC strain JG280 caused more death of pig intestinal IPEC-J2 cells than did 10⁹ CFU/ml, suggesting that ETEC-induced cell death was cell density dependent and that quorum sensing (QS) may play a role in pathogenesis. Subsequent investigations demonstrated a positive correlation between autoinducer 2 (AI-2) activity of JG280 and death of IPEC-J2 cells during the infection for up to 3 h. However, there was a negative correlation between AI-2 activity and expression of the JG280 enterotoxin genes estA and estB when IPEC-J2 cells were exposed to the pathogen at 10⁸ CFU/ml. We therefore cloned the luxS gene (responsible for AI-2 production) from JG280 and overexpressed it in E. coli DH5α, because deletion of the luxS gene was retarded by the lack of suitable antibiotic selection markers and the resistance of this pathogen to a wide range of antibiotics. The addition of culture fluid from E. coli DH5α with the overexpressed luxS reduced cell death of IPEC-J2 cells by 10⁸ CFU/ml JG280. The addition also reduced the estA expression by JG280. Nonpathogenic K88⁺ strain JFF4, which lacks the enterotoxin genes, caused no death of IPEC-J2 cells, although it produced AI-2 activity comparable to that produced by JG280. These results suggest the involvement of AI-2-mediated quorum sensing in K88⁺ ETEC pathogenesis, possibly through a negative regulation of STa production.