Responses of well-differentiated airway epithelial cell cultures from healthy donors and patients with cystic fibrosis to Burkholderia cenocepacia infection.
Well-differentiated cultures established from airway epithelia of patients with cystic fibrosis (CF cultures) exhibited goblet cell hyperplasia, increased secretion of mucus, and higher basal levels of interleukin-8 than similarly cultured cells from healthy donors. Upon apical infection with low doses (10(4) to 10(5) CFU) of Burkholderia cenocepacia isolate BC7, the two cultures gave different responses. While normal cultures trapped the added bacteria in the mucus layer, killed and/or inhibited bacterial replication, and prevented bacterial invasion of the cells, CF cultures failed to kill and/or supported the growth of bacteria, leading to invasion of underlying epithelial cells, compromised transepithelial permeability, and cell damage. Depletion of the surface mucus layer prior to bacterial infection rendered the normal cultures susceptible to bacterial invasion, but the invading bacteria were mainly confined to vacuoles within the cells and appeared to be nonviable. In contrast, bacteria that invaded cells in CF cultures were found free in the cytoplasm surrounded by intermediate filaments and also between cells. Cultured CF airway epithelium was therefore more susceptible to infection than normal epithelium. This mimics CF tissue in vivo and illustrates differences in the way epithelia in CF patients and normal subjects handle bacterial infection. In addition, we found that the CF and normal cell cultures responded differently not only to isolate BC7 but also to isolates of other B. cepacia complex species. We therefore conclude that this cell culture model is suitable for investigation of B. cepacia complex pathogenesis in CF patients.
Sajjan
Umadevi
U
Program in Lung Biology, Hospital For Sick Children, Toronto, Ontario, Canada. usajjan@umich.edu
Keshavjee
Shaf
S
Forstner
Janet
J
D019117:Burkholderia / Q000472:pathogenicity*; D019121:Burkholderia Infections / Q000378:metabolism*; D003550:Cystic Fibrosis / Q000382:microbiology*; D016207:Cytokines / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology; D005455:Fluorescent Antibody Technique; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism; D008854:Microscopy, Electron; D009077:Mucins / Q000378:metabolism; D009093:Mucus / Q000378:metabolism; D010539:Permeability; D012137:Respiratory System / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology
D016207:Cytokines; D016209:Interleukin-8; D009077:Mucins