Francisella tularensis invasion of lung epithelial cells.
Francisella tularensis, a gram-negative facultative intracellular bacterial pathogen, causes disseminating infections in humans and other mammalian hosts. Macrophages and other monocytes have long been considered the primary site of F. tularensis replication in infected animals. However, recently it was reported that F. tularensis also invades and replicates within alveolar epithelial cells following inhalation in a mouse model of tularemia. TC-1 cells, a mouse lung epithelial cell line, were used to study the process of F. tularensis invasion and intracellular trafficking within nonphagocytic cells. Live and paraformaldehyde-fixed F. tularensis live vaccine strain organisms associated with, and were internalized by, TC-1 cells at similar frequencies and with indistinguishable differences in kinetics. Inhibitors of microfilament and microtubule activity resulted in significantly decreased F. tularensis invasion, as did inhibitors of phosphatidylinositol 3-kinase and tyrosine kinase activity. Collectively, these results suggest that F. tularensis epithelial cell invasion is mediated by a preformed ligand on the bacterial surface and driven entirely by host cell processes. Once internalized, F. tularensis-containing endosomes associated with early endosome antigen 1 (EEA1) followed by lysosome-associated membrane protein 1 (LAMP-1), with peak coassociation frequencies occurring at 30 and 120 min postinoculation, respectively. By 2 h postinoculation, 70.0% (+/- 5.5%) of intracellular bacteria were accessible to antibody delivered to the cytoplasm, indicating vacuolar breakdown and escape into the cytoplasm.
Craven
Robin R
RR
Department of Microbiology and Immunology, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7290, USA.
Hall
Joshua D
JD
Fuller
James R
JR
Taft-Benz
Sharon
S
Kawula
Thomas H
TH
D000199:Actins / Q000378:metabolism; D000818:Animals; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D004705:Endocytosis; D011992:Endosomes / Q000382:microbiology; D004847:Epithelial Cells / Q000382:microbiology* / Q000648:ultrastructure; D005604:Francisella tularensis / Q000472:pathogenicity*; D006801:Humans; D008168:Lung / Q000166:cytology / Q000382:microbiology* / Q000648:ultrastructure; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000378:metabolism; D015398:Signal Transduction; D033921:Vesicular Transport Proteins / Q000378:metabolism; D052118:Lysosomal-Associated Membrane Protein 1
D000199:Actins; D051907:Lysosomal Membrane Proteins; D033921:Vesicular Transport Proteins; C052255:Lamp1 protein, mouse; D052118:Lysosomal-Associated Membrane Protein 1; C093846:early endosome antigen 1