Chronic respiratory infections by the Burkholderia cepacia complex (Bcc) are of great concern to patients with cystic fibrosis. Bcc isolates may survive intracellularly within amoebae, respiratory epithelial cells and macrophages. The molecular mechanisms facilitating colonization and pathogenesis remain unclear. Given the importance of bacterial adhesion to host surfaces in microbial pathogenesis, we investigated the role of the O antigen LPS in the interaction of Burkholderia cenocepacia, a member of the Bcc, with macrophages and epithelial cells. Our results demonstrated that the O antigen modulates phagocytosis but does not affect intracellular survival of B. cenocepacia. Internalization of strains that lack O antigen was significantly increased compared to that of their isogenic smooth counterparts. However, no differences between rough and smooth strains were found in their ability to delay phagosomal maturation. We also found that the O antigen interfered with the ability of B. cenocepacia to adhere to bronchial epithelial cells, suggesting that this polysaccharide may mask one or more bacterial surface adhesins.
ArjcharoenS.,
WikraiphatC.,
PudlaM.,
LimposuwanK.,
WoodsD. E.,
SirisinhaS.,
UtaisincharoenP.2007; Fate of a Burkholderia pseudomallei lipopolysaccharide mutant in the mouse macrophage cell line RAW 264.7: possible role for the O-antigenic polysaccharide moiety of lipopolysaccharide in internalization and intracellular survival. Infect Immun 75:4298–4304[CrossRef]
ClayC. D.,
SoniS.,
GunnJ. S.,
SchlesingerL. S.2008; Evasion of complement-mediated lysis and complement C3 deposition are regulated by Francisella tularensis lipopolysaccharide O antigen. J Immunol 181:5568–5578[CrossRef]
CoenyeT.,
VandammeP.2003; Diversity and significance of Burkholderia species occupying diverse ecological niches. Environ Microbiol 5:719–729[CrossRef]
CraigF. F.,
CooteJ. G.,
PartonR.,
FreerJ. H.,
GilmourN. J.1989; A plasmid which can be transferred between Escherichia coli and Pasteurella haemolytica by electroporation and conjugation. J Gen Microbiol 135:2885–2890
DesaiM.,
BuhlerT.,
WellerP. H.,
BrownM. R.1998; Increasing resistance to planktonic and biofilm cultures of Burkholderia cepacia to ciprofloxacin and ceftazidime during exponential growth. J Antimicrob Chemother 42:153–160[CrossRef]
FlannaganR. S.,
AubertD.,
KooiC.,
SokolP. A.,
ValvanoM. A.2007; Burkholderia cenocepacia requires a periplasmic HtrA protease for growth under thermal and osmotic stress and for survival in vivo. Infect Immun 75:1679–1689[CrossRef]
GovanJ. R. W.,
DereticV.1996; Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia
. Microbiol Rev 60:539–574
GovanJ. R.,
BrownP. H.,
MaddisonJ.,
DohertyC. J.,
NelsonJ. W.,
DoddM.,
GreeningA. P.,
WebbA. K.1993; Evidence for transmission of Pseudomonas cepacia by social contact in cystic fibrosis. Lancet 342:15–19[CrossRef]
JohnsonW. M.,
TylerS. D.,
RozeeK. R.1994; Linkage analysis of geographic and clinical clusters in Pseudomonas cepacia infections by multilocus enzyme electrophoresis and ribotyping. J Clin Microbiol 32:924–930
LamotheJ.,
HuynhK. K.,
GrinsteinS.,
ValvanoM. A.2007; Intracellular survival of Burkholderia cenocepacia in macrophages is associated with a delay in the maturation of bacteria-containing vacuoles. Cell Microbiol 9:40–53[CrossRef]
LesseA. J.,
CampagnariA. A.,
BittnerW. E.,
ApicellaM. A.1990; Increased resolution of lipopolysaccharides and lipooligosaccharides utilizing tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. J Immunol Methods 126:109–117[CrossRef]
LiPumaJ. J.,
SpilkerT.,
GillL. H.,
CampbellP. W.,
LiuL.,
MahenthiralingamE.2001; Disproportionate distribution of Burkholderia cepacia complex species and transmissibility markers in cystic fibrosis. Am J Respir Crit Care Med 164:92–96[CrossRef]
LoutetS. A.,
FlannaganR. S.,
KooiC.,
SokolP. A.,
ValvanoM. A.2006; A complete lipopolysaccharide inner core oligosaccharide is required for resistance of Burkholderia cenocepacia to polymyxin B and bacterial survival in vivo. J Bacteriol 188:2073–2080[CrossRef]
MaierT. M.,
CaseyM. S.,
BeckerR. H.,
DorseyC. W.,
GlassE. M.,
MaltsevN.,
ZahrtT. C.,
FrankD. W.2007; Identification of Francisella tularensis Himar1-based transposon mutants defective for replication in macrophages. Infect Immun 75:5376–5389[CrossRef]
MaroldaC. L.,
WelshJ.,
DafoeL.,
ValvanoM. A.1990; Genetic analysis of the O7-polysaccharide biosynthesis region from the Escherichia coli O7 : K1 strain VW187. J Bacteriol 172:3590–3599
MaroldaC. L.,
HauröderB.,
JohnM. A.,
MichelR.,
ValvanoM. A.1999; Intracellular survival and saprophytic growth of isolates from the Burkholderia cepacia complex in free-living amoebae. Microbiology 145:1509–1517[CrossRef]
MurrayG. L.,
AttridgeS. R.,
MoronaR.2006; Altering the length of the lipopolysaccharide O antigen has an impact on the interaction of Salmonella enterica serovar Typhimurium with macrophages and complement. J Bacteriol 188:2735–2739[CrossRef]
OrtegaX.,
HuntT. A.,
LoutetS.,
Vinion-DubielA. D.,
DattaA.,
ChoudhuryB.,
GoldbergJ. B.,
CarlsonR.,
ValvanoM. A.2005; Reconstitution of O-specific lipopolysaccharide expression in the Burkholderia cenocepacia strain J2315 that is associated with transmissible infections in patients with cystic fibrosis. J Bacteriol 187:1324–1333[CrossRef]
PorteF.,
NaroeniA.,
Ouahrani-BettacheS.,
LiautardJ. P.2003; Role of the Brucella suis lipopolysaccharide O antigen in phagosomal genesis and in inhibition of phagosome-lysosome fusion in murine macrophages. Infect Immun 71:1481–1490[CrossRef]
RajashekaraG.,
CovertJ.,
PetersenE.,
EskraL.,
SplitterG.2008; Genomic island 2 of Brucella melitensis is a major virulence determinant: functional analyses of genomic islands. J Bacteriol 190:6243–6252[CrossRef]
SainiL. S.,
GalsworthyS. B.,
JohnM. A.,
ValvanoM. A.1999; Intracellular survival of Burkholderia cepacia complex isolates in the presence of macrophage cell activation. Microbiology 145:3465–3475
SajjanS. U.,
ForstnerJ. F.1992; Identification of the mucin-binding adhesin of Pseudomonas cepacia isolated from patients with cystic fibrosis. Infect Immun 60:1434–1440
SajjanU. S.,
ForstnerJ. F.1993; Role of a 22-kilodalton pilin protein in binding of Pseudomonas cepacia to buccal epithelial cells. Infect Immun 61:3157–3163
SchäggerH.,
von JagowG.1987; Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166:368–379[CrossRef]
ViaL. E.,
FrattiR. A.,
McFaloneM.,
Pagan-RamosE.,
DereticD.,
DereticV.1998; Effects of cytokines on mycobacterial phagosome maturation. J Cell Sci 111:897–905