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Am. J. Respir. Cell Mol. Biol., Volume 25, Number 2, August, 2001 196-202

Antibacterial Activity of Apical Surface Fluid from the Human Airway Cell Line Calu-3
Pharmacologic Alteration by Corticosteroids and beta 2-Agonists

Yi Zhang, William W. Reenstra, and Aaron Chidekel

Departments of Medical and Clinical Research, and Pediatrics, Alfred I. duPont Hospital for Children, Wilmington, Delaware

Calu-3 cells, a human lung carcinoma cell line with properties like serous cells of the upper airway, were used to develop an in vitro model for airway antibacterial activity. Calu-3 cell monolayers were cultured on permeable supports at an air-liquid interface. Apical surface fluid (ASF) was collected by washing; antibacterial activity was assayed by incubating ASF washings with bacteria for 18 h and counting surviving colony-forming units. ASF washings killed Escherichia coli and Pseudomonas aeruginosa. Antibacterial activity was salt sensitive and dependent on protein concentration. After washing, approximately 30 h were required before antibacterial activity recovered to its initial level. After culturing with topical corticosteroids (budesonide, triamcinolone, or beclomethasone, 0.1 µg/ml for 48 h), ASF antibacterial activity was 4- to 10-fold greater than the ASF from control monolayers. The increase in antibacterial activity was dose-dependent. The beta 2-agonists salbutamol and terbutaline (100 µg/ml for 48 h) decreased ASF antibacterial activity by 5- to 8-fold. The nonsteroidal anti-inflammatory agents ibuprofen and cromolyn sodium had no effect. Our results are most consistent with agonist-dependent changes in the composition of ASF antibacterial proteins. We conclude that Calu-3 cells synthesize and secrete antibacterial proteins and that clinical agents can alter these functions.

Abbreviations: apical surface fluid, ASF; complementary DNA, cDNA; cystic fibrosis, CF; cystic fibrosis transmembrane conductance regulator, CFTR; colony-forming unit, CFU; Luria broth, LB; messenger RNA, mRNA; nuclear factor kappa B, NF-kappa B; reverse transcriptase/polymerase chain reaction, RT-PCR; sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE; standard error of the mean, SEM; transepithelial resistance, TER.




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