title: A type IV secretion system contributes to intracellular survival and replication of Burkholderia cenocepacia.
abstract: Burkholderia cenocepacia is an important respiratory pathogen in persons with cystic fibrosis (CF). Recent studies indicate that B. cenocepacia survives within macrophages and airway epithelial cells in vitro by evading endosome-lysosome fusion. We investigated the role of a plasmid-encoded type IV secretion system in the intracellular survival, replication, and processing of B. cenocepacia. Both a wild-type strain (K56-2) and its type IV secretion system mutant (designated LC101) entered and replicated in CF airway epithelial cells and monocyte-derived macrophages. However, significantly more intracellular K56-2 than LC101 bacteria were found in both cell types at 24 h postinfection. Colocalization of bacteria with markers of the classical endocytic pathway indicated that although both K56-2 and LC101 reside transiently in early endosomes, a greater proportion of the mutant bacteria are targeted to lysosomal degradation. In contrast, wild-type bacteria escape from the classical endocytic pathway and traffic to the endoplasmic reticulum, where they replicate. Our results show that the intracellular processing of B. cenocepacia is similar in both professional and nonprofessional phagocytes and that a functional plasmid-encoded type IV secretion system contributes to the survival and replication of B. cenocepacia in eukaryotic cells.
authors.affiliation: Department of Pediatrics and Communicable Diseases, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
authors.forename: S Umadevi
authors.initials: SU
authors.lastname: Sajjan
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Carmody
authors.forename: Carlos F
authors.initials: CF
authors.lastname: Gonzalez
authors.forename: John J
authors.initials: JJ
authors.lastname: LiPuma
mesh_terms: D015139:Blotting, Southern; D019121:Burkholderia Infections / Q000235:genetics / Q000378:metabolism*; D042602:Burkholderia cepacia complex / Q000472:pathogenicity* / Q000502:physiology*; D002460:Cell Line; D003550:Cystic Fibrosis / Q000382:microbiology; D004721:Endoplasmic Reticulum; D011992:Endosomes / Q000382:microbiology; D004847:Epithelial Cells / Q000382:microbiology; D005455:Fluorescent Antibody Technique; D006801:Humans; D008247:Lysosomes / Q000382:microbiology; D008264:Macrophages / Q000382:microbiology; D018613:Microscopy, Confocal; D016254:Mutagenesis, Insertional; D020133:Reverse Transcriptase Polymerase Chain Reaction