Influence of FkpA variants on survival and replication of Cronobacter spp. in human macrophages.
Members of the genus Cronobacter are responsible for cases of meningitis and bacteremia with high fatality rates in neonates. Macrophage uptake of invading microbes is an innate process, and it has been proposed that macrophage infectivity potentiator (Mip) like proteins enhance the ability of pathogens to survive within macrophages. Cronobacter harbor the mip-like gene fkpA, but its role in intracellular survival of these bacteria in human macrophages has not yet been studied. Application of gentamicin exclusion assays and human THP-1 macrophage cells revealed significant differences in the capablility of Cronobacter species to survive and replicate within macrophages. Analysis to the amino acid level revealed both length and sequence variations in FkpA proteins among species. In this study, we addressed the possible influence of FkpA variants in intracellular survival of Cronobacter spp. in human macrophages, by knocking out the fkpA genes in two different Cronobacter strains and subsequent complementation with variants of the fkpA genes. Our results provide strong evidence that, in Cronobacter spp., FkpA must be considered a virulence factor, but its influence on macrophage survival and replication varies among strains and/or species due to the presence of amino acid variations.
Institute for Food Safety and Hygiene, Vetsuisse Faculty, University of Zurich, Winterthurerstr. 272, CH-8057 Zurich, Switzerland. Electronic address: athmanya.eshwar@uzh.ch.
Athmanya Konegadde
AK
Eshwar
Institute for Food Safety and Hygiene, Vetsuisse Faculty, University of Zurich, Winterthurerstr. 272, CH-8057 Zurich, Switzerland. Electronic address: tasarat@fsafety.uzh.ch.
Taurai
T
Tasara
Institute for Food Safety and Hygiene, Vetsuisse Faculty, University of Zurich, Winterthurerstr. 272, CH-8057 Zurich, Switzerland. Electronic address: stephanr@fsafety.uzh.ch.
Roger
R
Stephan
Institute for Food Safety and Hygiene, Vetsuisse Faculty, University of Zurich, Winterthurerstr. 272, CH-8057 Zurich, Switzerland. Electronic address: lehnera@fsafety.uzh.ch.
Angelika
A
Lehner
D002461:Cell Line, Transformed; D059126:Cronobacter / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006801:Humans; D008264:Macrophages / Q000382:microbiology* / Q000648:ultrastructure; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D037521:Virulence Factors / Q000235:genetics
D037521:Virulence Factors
CLSM; Cronobacter spp.; FkpA; Macrophage survival; THP-1; Virulence factor