Decreased apoptosis in the ileum and ileal Peyer's patches: a feature after infection with rabbit enteropathogenic Escherichia coli O103.
Significant changes occur in intestinal epithelial cells after infection with enteropathogenic Escherichia coli (EPEC). However, it is unclear whether this pathogen alters rates of apoptosis. By using a naturally occurring weaned rabbit infection model, we determined physiological levels of apoptosis in rabbit ileum and ileal Peyer's patches (PP) and compared them to those found after infection with adherent rabbit EPEC (REPEC O103). Various REPEC O103 strains were first tested in vitro for characteristic virulence features. Rabbits were then inoculated with the REPEC O103 strains that infected cultured cells the most efficiently. After experimental infection, intestinal samples were examined by light and electron microscopy. Simultaneously, ileal apoptosis was assessed by using terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) and caspase 3 assays and by apoptotic cell counts based on morphology (hematoxylin-and-eosin staining). The highest physiological apoptotic indices were measured in PP germinal centers (median = 14.7%), followed by PP domed villi (8.1%), tips of absorptive villi (3.8%), and ileal crypt regions (0.5%). Severe infection with REPEC O103 resulted in a significant decrease in apoptosis in PP germinal centers (determined by TUNEL assay; P = 0.01), in the tips of ileal absorptive villi (determined by H&E staining; P = 0.04), and in whole ileal cell lysates (determined by caspase 3 assay; P = 0.001). We concluded that REPEC O103 does not promote apoptosis. Furthermore, we cannot rule out the possibility that REPEC O103, in fact, decreases apoptotic levels in the rabbit ileum.
Heczko
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Biotechnology Laboratory and Departments of Microbiology and Immunology, Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
Carthy
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O'Brien
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Finlay
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D000818:Animals; D017209:Apoptosis; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D004269:DNA, Bacterial; D004755:Enterobacteriaceae / Q000502:physiology; D004926:Escherichia coli / Q000378:metabolism / Q000472:pathogenicity; D004927:Escherichia coli Infections / Q000378:metabolism / Q000473:pathology*; D029968:Escherichia coli Proteins; D006367:HeLa Cells; D006801:Humans; D007082:Ileum / Q000473:pathology* / Q000648:ultrastructure; D020287:In Situ Nick-End Labeling; D008565:Membrane Proteins / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D010581:Peyer's Patches / Q000473:pathology* / Q000648:ultrastructure; D010957:Plasmids; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000378:metabolism; D022621:Shiga Toxin / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods; D014774:Virulence
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C087863:EaeB protein, E coli; D029968:Escherichia coli Proteins; C102051:EspA protein, E coli; C474481:EspD protein, E coli; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; C109287:Tir protein, E coli; D022621:Shiga Toxin; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases