Bacteriophage EK99P-1 alleviates enterotoxigenic Escherichia coli K99-induced barrier dysfunction and inflammation

Abstract

Bacteriophages, simply phages, have long been used as a potential alternative to antibiotics for livestock due to their ability to specifically kill enterotoxigenic Escherichia coli (ETEC), which is a major cause of diarrhea in piglets. However, the control of ETEC infection by phages within intestinal epithelial cells, and their relationship with host immune responses, remain poorly understood. In this study, we evaluated the effect of phage EK99P-1 against ETEC K99-infected porcine intestinal epithelial cell line (IPEC-J2). Phage EK99P-1 prevented ETEC K99-induced barrier disruption by attenuating the increased permeability mediated by the loss of tight junction proteins such as zonula occludens-1 (ZO-1), occludin, and claudin-3. ETEC K99-induced inflammatory responses, such as interleukin (IL)-8 secretion, were decreased by treatment with phage EK99P-1. We used a IPEC-J2/peripheral blood mononuclear cell (PBMC) transwell co-culture system to investigate whether the modulation of barrier disruption and chemokine secretion by phage EK99P-1 in ETEC K99-infected IPEC-J2 would influence immune cells at the site of basolateral. The results showed that phage EK99P-1 reduced the mRNA expression of ETEC K99-induced pro-inflammatory cytokines, IL-1β and IL-8, from PBMC collected on the basolateral side. Together, these results suggest that phage EK99P-1 prevented ETEC K99-induced barrier dysfunction in IPEC-J2 and alleviated inflammation caused by ETEC K99 infection. Reinforcement of the intestinal barrier, such as regulation of permeability and cytokines, by phage EK99P-1 also modulates the immune cell inflammatory response.

Whole Genome Sequencing and CRISPR/Cas9 Gene Editing of Enterotoxigenic Escherichia coli BE311 for Fluorescence Labeling and Enterotoxin Analyses

“…Enterotoxigenic Escherichia coli (ETEC) is one of the main causes of diarrhea in farm animals, including piglets, calves, and lambs [ 1 ], and infections by ETEC in animal husbandry and aquaculture result in severe economic losses due to high animal morbidity and mortality [ 2 ]. Generally, ETEC in an exogenous environment enters animals orally and is subsequently recognized by specific receptors in the small intestine.…”

Effect of phage vB_EcoM_FJ1 on the reduction of ETEC O9:H9 infection in a neonatal pig cell line

“…Under the same pH as that of the piglet intestine, successful indicators were obtained: the microencapsulated FJ1 decreased EC43 load in 3.9 Log CFU cm −2 (> 99.9%), also 6 h after phage administration (Figure 3 ). Phage effectiveness in decreasing bacterial contamination in intestinal cell lines was previously observed by Mirzaei et al [ 36 ] in HT-29 and Caco-2, with reductions of 1.0 to 4.0 Log CFU mL −1 of ESBL E. coli strains for 8 h. More recently, Kim et al [ 37 ], reported that the phage EK99P-1 was able to alleviate ETEC K99-induced barrier dysfunction and inflammatory response of IPEC-J2 cultured cells. In this study, the protective effect, that FJ1 seems to have on EC43-infected cells (at least 16 h after phage administration), was observed microscopically, showing an apparent delay in cell damage.…”

Potential Probiotic Lacticaseibacillus paracasei MJM60396 Prevents Hyperuricemia in a Multiple Way by Absorbing Purine, Suppressing Xanthine Oxidase and Regulating Urate Excretion in Mice

“…Furthermore, MJM60396 showed strong antibacterial activity against many enteric pathogens ( Table S2 ). These enteric pathogens induce intestinal toxins and cause diarrhea, and destroy the intestinal barrier by causing loss of tight junction proteins such as ZO-1 and occludin [ 47 ]. MJM60396 showed an antibacterial effect on enteric pathogens, thus protecting the intestinal barrier.…”