title: Recruitment of dendritic cells is responsible for intestinal epithelial damage in the pathogenesis of necrotizing enterocolitis by Cronobacter sakazakii.
abstract: Cronobacter sakazakii is a Gram-negative pathogen associated with the cases of necrotizing enterocolitis (NEC) that result from formula contamination. In a mouse model of NEC, we demonstrate that C. sakazakii infection results in epithelial damage by recruiting greater numbers of dendritic cells (DCs) than macrophages and neutrophils in the gut and suppresses DC maturation, which requires outer membrane protein A (OmpA) expression in C. sakazakii. Pretreatment of intestinal epithelial cell monolayers with supernatant from OmpA(+) C. sakazakii/DC culture markedly enhanced membrane permeability and enterocyte apoptosis, whereas OmpA(-) C. sakazakii/DC culture supernatant had no effect. Analysis of OmpA(+) C. sakazakii/DC coculture supernatant revealed significantly greater TGF-β production compared with the levels produced by OmpA(-) C. sakazakii infection. TGF-β levels were elevated in the intestinal tissue of mice infected with OmpA(+) C. sakazakii. Cocultures of CaCo-2 cells and DCs in a "double-layer" model followed by infection with OmpA(+) C. sakazakii significantly enhanced monolayer leakage by increasing TGF-β production. Elevated levels of inducible NO synthase (iNOS) were also observed in the double-layer infection model, and abrogation of iNOS expression prevented the C. sakazakii-induced CaCo-2 cell monolayer permeability despite the presence of DCs or OmpA(+) C. sakazakii/DC supernatant. Blocking TGF-β activity using a neutralizing Ab suppressed iNOS production and prevented apoptosis and monolayer leakage. Depletion of DCs in newborn mice protected against C. sakazakii-induced NEC, whereas adoptive transfer of DCs rendered the animals susceptible to infection. Therefore, C. sakazakii interaction with DCs in intestine enhances the destruction of the intestinal epithelium and the onset of NEC due to increased TGF-β production.
authors.affiliation: Department of Surgery, The Saban Research Institute, Children's Hospital Los Angeles, Los Angeles, CA 90027, USA.
authors.forename: Claudia N
authors.initials: CN
authors.lastname: Emami
authors.forename: Rahul
authors.initials: R
authors.lastname: Mittal
authors.forename: Larry
authors.initials: L
authors.lastname: Wang
authors.forename: Henri R
authors.initials: HR
authors.lastname: Ford
authors.forename: Nemani V
authors.initials: NV
authors.lastname: Prasadarao
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000502:physiology; D018938:Caco-2 Cells; D018920:Coculture Techniques; D044083:Cronobacter sakazakii / Q000472:pathogenicity*; D003713:Dendritic Cells / Q000382:microbiology / Q000473:pathology*; D020345:Enterocolitis, Necrotizing / Q000209:etiology* / Q000382:microbiology / Q000473:pathology; D006801:Humans; D007413:Intestinal Mucosa / Q000382:microbiology / Q000473:pathology*; D051379:Mice; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D016212:Transforming Growth Factor beta; C078167:OMPA outer membrane proteins; D052247:Nitric Oxide Synthase Type II