Entry of Burkholderia organisms into respiratory epithelium: CFTR, microfilament and microtubule dependence.
BACKGROUND
The pathogenesis of infection with Burkholderia cepacia complex (Bcc) organisms may be linked to its capacity to invade respiratory epithelium.


METHODS
An antibiotic exclusion assay was used to study B. dolosa AU4459 and B. cenocepacia J2315 invasion into wild-type (WT) and CFTR-deficient respiratory epithelial cells. Inhibitors were used to evaluate Bcc invasion dependency on host microtubule (mt) and microfilament (mf) systems.


RESULTS
B. dolosa entered WT-CFTR cells with 5-fold greater efficiency than CFTR deficient cells (25% vs 5%, respectively). Invasion dropped to <0.5% after either mf or mt inhibition. B. cenocepacia entered WT (0.05%) and CFTR-deficient cells (0.07%) with similarly low efficiencies, which significantly decreased with either mf or mt inhibition (0.008% and 0.002%, respectively).


CONCLUSIONS
B. dolosa and B. cenocepacia enter respiratory epithelial cells in a mf and mt dependent fashion. Mutated CFTR leads to less internalization of B. dolosa, but not B. cenocepacia.
Taylor
Jane B
JB
Division of Allergy, Immunology and Pulmonary Medicine, Department of Pediatrics, St. Louis Children's Hospital, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Hogue
Lisa A
LA
LiPuma
John J
JJ
Walter
Michael J
MJ
Brody
Steven L
SL
Cannon
Carolyn L
CL
D008841:Actin Cytoskeleton / Q000502:physiology; D000583:Amikacin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D019121:Burkholderia Infections / Q000188:drug therapy / Q000382:microbiology* / Q000473:pathology; D042725:Burkholderia gladioli / Q000254:growth & development* / Q000472:pathogenicity* / Q000648:ultrastructure; D002442:Ceftazidime / Q000494:pharmacology; D002461:Cell Line, Transformed; D003550:Cystic Fibrosis / Q000382:microbiology* / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004359:Drug Therapy, Combination; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology* / Q000502:physiology; D006801:Humans; D000077731:Meropenem; D008826:Microbial Sensitivity Tests; D046529:Microscopy, Electron, Transmission; D008870:Microtubules / Q000502:physiology; D020545:Respiratory Mucosa / Q000166:cytology / Q000382:microbiology*; D013845:Thienamycins / Q000494:pharmacology; D014774:Virulence
D000900:Anti-Bacterial Agents; C505032:CFTR protein, human; D013845:Thienamycins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000583:Amikacin; D002442:Ceftazidime; D000077731:Meropenem