ABCB1 (P-glycoprotein) reduces bacterial attachment to human gastrointestinal LS174T epithelial cells.
The aim of this project was to show elevated P-glycoprotein (P-gp) expression decreasing bacterial association with LS174T human gastrointestinal cells, and that this effect could be reversed upon blocking functional P-gp efflux. Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Lactobacillus acidophilus and numerous strains of Escherichia coli, from commensal to enteropathogenic and enterohaemorrhagic strains (O157:H7) were fluorescently labelled and incubated on LS174T cultures either with or without P-gp amplification using rifampicin. PSC-833 was used as a potent functional P-gp blocking agent. Staphylococcus and Pseudomonas displayed the greatest association with the LS174T cells. Surprisingly, lactobacilli retained more fluorescence than enteropathogenic-E. coli in this system. Irrespective of attachment differences between the bacterial species, the increase in P-gp protein expression decreased bacterial fluorescence by 25-30%. This included the GFP-labelled E. coli, and enterohaemorrhagic E. coli (O157:H7). Blocking P-gp function through the co-administration of PSC-833 increased the amount of bacteria associated with P-gp expressing LS174T cells back to control levels. As most bacteria were affected to the same degree, irrespective of pathogenicity, it is unlikely that P-gp has a direct influence on adhesion of bacteria, and instead P-gp may be playing an indirect role by secreting a bank of endogenous factors or changing the local environment to one less suited to bacterial growth in general.
School of Pharmacy, Curtin University and Curtin Health Innovation Research Institute (CHIRI), Perth, Western Australia, Australia. a.p.crowe@curtin.edu.au
Andrew
A
Crowe
Mary
M
Bebawy
D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D001287:Attachment Sites, Microbiological / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D004847:Epithelial Cells / Q000378:metabolism* / Q000382:microbiology* / Q000473:pathology; D019453:Escherichia coli O157 / Q000378:metabolism; D041981:Gastrointestinal Tract / Q000378:metabolism* / Q000382:microbiology* / Q000473:pathology; D006801:Humans; D007711:Klebsiella pneumoniae / Q000378:metabolism; D007779:Lactobacillus acidophilus / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000378:metabolism; D013211:Staphylococcus aureus / Q000378:metabolism
C513055:ABCB1 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1