Penetration of clinical isolates of Pseudomonas aeruginosa through MDCK epithelial cell monolayers.
Pseudomonas aeruginosa causes both invasive (bacteremic) and chronic noninvasive infections. A simple in vitro system to screen P. aeruginosa clinical isolates for their capacity to penetrate MDCK cell monolayers has been developed. By means of this system, P. aeruginosa clinical isolates, including 32 blood and 45 respiratory isolates, were examined. When monolayers were infected with 3.5x107 cfu of bacteria, significantly more blood (93.7%) than respiratory (54.4%) isolates (P<.001) were detected in the basolateral medium after 3 h. Penetration ability was usually independent of cytotoxicity. Only 8 (4 blood and 4 respiratory) isolates were cytotoxic, possessed exoU, and passed through the monolayer after epithelial cell death, associated with a marked drop in transepithelial electrical resistance. Conversely, noncytotoxic isolates with high penetration ability but without severe epithelial damage were invasive. This system is well suited for screening clinical isolates and their mutants for specific genes conferring the invasiveness phenotype.
Hirakata
Y
Y
Division of Infectious and Immunological Diseases, Department of Pediatrics, University of British Columbia, Vancouver, Canada V5Z 4H4. hirakata@net.nagasaki-u.ac.jp
Finlay
B B
BB
Simpson
D A
DA
Kohno
S
S
Kamihira
S
S
Speert
D P
DP
D000818:Animals; D001431:Bacteriological Techniques; D002460:Cell Line; D002470:Cell Survival; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology*; D004926:Escherichia coli / Q000502:physiology; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000472:pathogenicity* / Q000502:physiology
D007770:L-Lactate Dehydrogenase