title: Putative Inv is essential for basolateral invasion of Caco-2 cells and acts synergistically with OmpA to affect in vitro and in vivo virulence of Cronobacter sakazakii ATCC 29544.
abstract: Cronobacter sakazakii is an opportunistic pathogen that causes neonatal meningitis and necrotizing enterocolitis. Its interaction with intestinal epithelium is important in the pathogenesis of enteric infections. In this study, we investigated the involvement of the inv gene in the virulence of C. sakazakii ATCC 29544 in vitro and in vivo. Sequence analysis of C. sakazakii ATCC 29544 inv revealed that it is different from other C. sakazakii isolates. In various cell culture models, an Δinv deletion mutant showed significantly lowered invasion efficiency, which was restored upon genetic complementation. Studying invasion potentials using tight-junction-disrupted Caco-2 cells suggested that the inv gene product mediates basolateral invasion of C. sakazakii ATCC 29544. In addition, comparison of invasion potentials of double mutant (ΔompA Δinv) and single mutants (ΔompA and Δinv) provided evidence for an additive effect of the two putative outer membrane proteins. Finally, the importance of inv and the additive effect of putative Inv and OmpA were also proven in an in vivo rat pup model. This report is the first to demonstrate two proteins working synergistically in vitro, as well as in vivo in C. sakazakii pathogenesis.
authors.affiliation: Department of Food Science and Technology, College of Agriculture and Life Sciences, Chonbuk National University, Jeonju, South Korea.
authors.forename: Dilini
authors.initials: D
authors.lastname: Chandrapala
authors.forename: Kyumson
authors.initials: K
authors.lastname: Kim
authors.forename: Younho
authors.initials: Y
authors.lastname: Choi
authors.forename: Amal
authors.initials: A
authors.lastname: Senevirathne
authors.forename: Dong-Hyun
authors.initials: DH
authors.lastname: Kang
authors.forename: Sangryeol
authors.initials: S
authors.lastname: Ryu
authors.forename: Kwang-Pyo
authors.initials: KP
authors.lastname: Kim
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002460:Cell Line; D044083:Cronobacter sakazakii / Q000145:classification* / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D006801:Humans; D051381:Rats; D017207:Rats, Sprague-Dawley; D013047:Specific Pathogen-Free Organisms; D014774:Virulence
chemical_list: D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; C078167:OMPA outer membrane proteins