TY - JOUR
T1 - Secretion of Alpha-Hemolysin by Escherichia coli Disrupts Tight Junctions in Ulcerative Colitis Patients
AU - Mirsepasi-lauridsen, Hengameh Chloé
AU - Du, Zhengyu
AU - Struve, Carsten
AU - Charbon, Godefroid
AU - Karczewski, Jurgen
AU - Krogfelt, Karen Angeliki
AU - Petersen, Andreas Munk
AU - Wells, Jerry M.
PY - 2016/3/1
Y1 - 2016/3/1
N2 - OBJECTIVES:
The potential of Escherichia coli (E. coli) isolated from inflammatory bowel disease (IBD) patients to damage the integrity of the intestinal epithelium was investigated.
METHODS:
E. coli strains isolated from patients with ulcerative colitis (UC) and healthy controls were tested for virulence capacity by molecular techniques and cytotoxic assays and transepithelial electric resistance (TER). E. coliisolate p19A was selected, and deletion mutants were created for alpha-hemolysin (α-hemolysin) (hly) clusters and cytotoxic necrotizing factor type 1 (cnf1). ProbioticE. coliNissle and pathogenicE. coliLF82 were used as controls.
RESULTS:
E. colistrains from patients with active UC completely disrupted epithelial cell tight junctions shortly after inoculation. These strains belong to phylogenetic group B2 and are all α-hemolysin positive. In contrast, probioticE. coliNissle, pathogenicE. coliLF82, fourE. colifrom patients with inactive UC and threeE. colistrains from healthy controls did not disrupt tight junctions.E. colip19A WT as well ascnf1, and single loci ofhlymutants from cluster I and II were all able to damage Caco-2 (Heterogeneous human epithelial colorectal adenocarcinoma) cell tight junctions. However, this phenotype was lost in a mutant with knockout (Δ) of bothhlyloci (P<0.001).
CONCLUSIONS:
UC-associated E. coliproducing α-hemolysin can cause rapid loss of tight junction integrity in differentiated Caco-2 cell monolayers. This effect was abolished in a mutant unable to express α-hemolysin. These results suggest that high Hly expression may be a mechanism by which specific strains of E. colipathobionts can contribute to epithelial barrier dysfunction and pathophysiology of disease in IBD.
AB - OBJECTIVES:
The potential of Escherichia coli (E. coli) isolated from inflammatory bowel disease (IBD) patients to damage the integrity of the intestinal epithelium was investigated.
METHODS:
E. coli strains isolated from patients with ulcerative colitis (UC) and healthy controls were tested for virulence capacity by molecular techniques and cytotoxic assays and transepithelial electric resistance (TER). E. coliisolate p19A was selected, and deletion mutants were created for alpha-hemolysin (α-hemolysin) (hly) clusters and cytotoxic necrotizing factor type 1 (cnf1). ProbioticE. coliNissle and pathogenicE. coliLF82 were used as controls.
RESULTS:
E. colistrains from patients with active UC completely disrupted epithelial cell tight junctions shortly after inoculation. These strains belong to phylogenetic group B2 and are all α-hemolysin positive. In contrast, probioticE. coliNissle, pathogenicE. coliLF82, fourE. colifrom patients with inactive UC and threeE. colistrains from healthy controls did not disrupt tight junctions.E. colip19A WT as well ascnf1, and single loci ofhlymutants from cluster I and II were all able to damage Caco-2 (Heterogeneous human epithelial colorectal adenocarcinoma) cell tight junctions. However, this phenotype was lost in a mutant with knockout (Δ) of bothhlyloci (P<0.001).
CONCLUSIONS:
UC-associated E. coliproducing α-hemolysin can cause rapid loss of tight junction integrity in differentiated Caco-2 cell monolayers. This effect was abolished in a mutant unable to express α-hemolysin. These results suggest that high Hly expression may be a mechanism by which specific strains of E. colipathobionts can contribute to epithelial barrier dysfunction and pathophysiology of disease in IBD.
U2 - 10.1038/ctg.2016.3
DO - 10.1038/ctg.2016.3
M3 - Article
SN - 2155-384X
VL - 7
SP - e149
JO - Clinical and Translational Gastroenterology
JF - Clinical and Translational Gastroenterology
IS - 3
ER -
