Campylobacter jejuni cocultured with epithelial cells reduces surface capsular polysaccharide expression.
The host cell environment can alter bacterial pathogenicity. We employed a combination of cellular and molecular techniques to study the expression of Campylobacter jejuni polysaccharides cocultured with HCT-8 epithelial cells. After two passages, the amount of membrane-bound high-molecular-weight polysaccharide was considerably reduced. Microarray profiling confirmed significant downregulation of capsular polysaccharide (CPS) locus genes. Experiments using conditioned media showed that sugar depletion occurred only when the bacterial and epithelial cells were cocultured. CPS depletion occurred when C. jejuni organisms were exposed to conditioned media from a different C. jejuni strain but not when exposed to conditioned media from other bacterial species. Proteinase K or heat treatment of conditioned media under coculture conditions abrogated the effect on the sugars, as did formaldehyde fixation and cycloheximide treatment of host cells or chloramphenicol treatment of the bacteria. However, sugar depletion was not affected in flagellar export (fliQ) and quorum-sensing (luxS) gene mutants. Passaged C. jejuni showed reduced invasiveness and increased serum sensitivity in vitro. C. jejuni alters its surface polysaccharides when cocultured with epithelial cells, suggesting the existence of a cross talk mechanism that modulates CPS expression during infection.
Corcionivoschi
N
N
The Children's Research Centre, Our Lady's Children's Hospital, Crumlin, Dublin 12, Ireland.
Clyne
M
M
Lyons
A
A
Elmi
A
A
Gundogdu
O
O
Wren
B W
BW
Dorrell
N
N
Karlyshev
A V
AV
Bourke
B
B
D016123:Campylobacter jejuni / Q000502:physiology*; D002460:Cell Line; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D004847:Epithelial Cells / Q000382:microbiology*; D020869:Gene Expression Profiling; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis*
D017077:Culture Media, Conditioned; D011135:Polysaccharides, Bacterial