title: Cronobacter sakazakii clinical isolates overcome host barriers and evade the immune response.
abstract: Cronobacter sakazakii is the most frequently clinically isolated species of the Cronobacter genus. However the virulence factors of C. sakazakii including their ability to overcome host barriers remains poorly studied. In this study, ten clinical isolates of C. sakazakii were assessed for their ability to invade and translocate through human colonic carcinoma epithelial cells (Caco-2) and human brain microvascular endothelial cells (HBMEC). Their ability to avoid phagocytosis in human macrophages U937 and human brain microglial cells was investigated. Additionally, they were tested for serum sensitivity and the presence of the Cronobacter plasminogen activation gene (cpa) gene, which is reported to confer serum resistance. Our data showed that the clinical C. sakazakii strains invaded and translocated through Caco-2 and HBMEC cell lines and some strains showed significantly higher levels of invasion and translocation. Moreover, C. sakazakii was able to persist and even multiply in phagocytic macrophage and microglial cells. All strains, except one, were able to withstand human serum exposure, the single serum sensitive strain was also the only one which did not encode for the cpa gene. These results demonstrate that C. sakazakii clinical isolates are able to overcome host barriers and evade the host immune response indicating their capacity to cause diseases such as necrotizing enterocolitis (NEC) and meningitis. Our data showed for the first time the ability of C. sakazakii clinical isolates to survive and multiply within human microglial cells. Additionally, it was shown that C. sakazakii clinical strains have the capacity to translocate through the Caco-2 and HBMEC cell lines paracellularly.
authors.affiliation: Pathogen Research Group, School of Science and Technology, Nottingham Trent University, Clifton Lane, Nottingham NG 11 8NS, UK; College of Applied Medical Sciences, King Saud bin Abdulaziz University for Health Sciences, Riyadh 11426, Saudi Arabia.
authors.forename: Faisal S
authors.initials: FS
authors.lastname: Almajed
authors.affiliation: Pathogen Research Group, School of Science and Technology, Nottingham Trent University, Clifton Lane, Nottingham NG 11 8NS, UK. Electronic address: Stephen.forsythe@ntu.ac.uk.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Forsythe
mesh_terms: D018938:Caco-2 Cells; D018929:Cell Culture Techniques; D002460:Cell Line; D044083:Cronobacter sakazakii / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification / Q000472:pathogenicity*; D042783:Endothelial Cells / Q000382:microbiology; D004756:Enterobacteriaceae Infections / Q000235:genetics / Q000276:immunology* / Q000382:microbiology*; D004847:Epithelial Cells / Q000382:microbiology; D006801:Humans; D008264:Macrophages / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D008581:Meningitis / Q000382:microbiology; D008826:Microbial Sensitivity Tests; D017628:Microglia / Q000382:microbiology / Q000473:pathology; D037521:Virulence Factors / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D037521:Virulence Factors
keywords: Cronobacter sakazakii; Meningitis; Microglia; Necrotising enterocolitis