title: Pathological and therapeutic significance of cellular invasion by Proteus mirabilis in an enterocystoplasty infection stone model.
abstract: Proteus mirabilis infection often leads to stone formation. We evaluated how bacterium-mucin adhesion, invasion, and intracellular crystal formation are related to antibiotic sensitivity and may cause frequent stone formation in enterocystoplasties. Five intestinal (Caco-2, HT29, HT29-18N2, HT29-FU, and HT29-MTX) and one ureter cell line (SV-HUC-1) were incubated in artificial urine with five Proteus mirabilis strains. Fluorescence-activated cell sorting (FACS), laser scanning microscopy, and electron microscopy evaluated cellular adhesion and/or invasion, pathologic changes to mitochondria, and P. mirabilis-mucin colocalization (MUC2 and MUC5AC). An MTT (thiazolyl blue tetrazolium bromide) assay and FACS analysis of caspase-3 evaluated the cellular response. Infected cells were incubated with antibiotics at dosages representing the expected urinary concentrations in a 10-year-old, 30-kg child to evaluate bacterial invasion and survival. All cell lines showed colocalization of P. mirabilis with human colonic mucin (i.e., MUC2) and human gastric mucin (i.e., MUC5AC). The correlation between membrane mucin expression and invasion was significant and opposite for SV-HUC-1 and HT29-MTX. Microscopically, invasion by P. mirabilis with intracellular crystal formation and mitochondrial damage was found. Double membranes surrounded bacteria in intestinal cells. Relative resistance to cotrimoxazole and augmentin was found in the presence of epithelial cells. Ciprofloxacin and gentamicin remained effective. Membrane mucin expression was correlated with relative antibiotic resistance. Cell invasion by P. mirabilis and mucin- and cell type-related distribution and response differences indicate bacterial tropism that affects crystal formation and mucosal presence. Bacterial invasion seems to have cell type-dependent mechanisms and prolong bacterial survival in antibiotic therapy, giving a new target for therapeutic optimalization of antibiotic treatment.
authors.affiliation: Subdivision of Pediatric Urology, Department of Urology, Sophia Children's Hospital, Erasmus Medical Center, 3000 DR Rotterdam, The Netherlands. rbmathoera@hotmail.com
authors.forename: Rejiv B
authors.initials: RB
authors.lastname: Mathoera
authors.forename: Dik J
authors.initials: DJ
authors.lastname: Kok
authors.forename: Cees M
authors.initials: CM
authors.lastname: Verduin
authors.forename: Rien J M
authors.initials: RJ
authors.lastname: Nijman
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001422:Bacterial Adhesion; D002460:Cell Line; D002648:Child; D003470:Culture Media; D024881:Drug Resistance, Bacterial; D006801:Humans; D007422:Intestines / Q000166:cytology / Q000382:microbiology*; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D009077:Mucins / Q000378:metabolism; D011512:Proteus Infections / Q000382:microbiology; D011513:Proteus mirabilis / Q000187:drug effects* / Q000472:pathogenicity*; D014513:Ureter / Q000166:cytology / Q000382:microbiology*; D014545:Urinary Calculi; D014556:Urine / Q000382:microbiology
chemical_list: D000900:Anti-Bacterial Agents; D003470:Culture Media; D009077:Mucins