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Am. J. Respir. Cell Mol. Biol., Volume 25, Number 6, December, 2001 699-706

Effects of Surfactant Protein A and NaCl Concentration on the Uptake of Pseudomonas aeruginosa by THP-1 Cells

Kavita R. Khubchandani, Rebecca E. Oberley, and Jeanne M. Snyder

Department of Anatomy and Cell Biology, University of Iowa College of Medicine, Iowa City, Iowa

In the present study, we characterized a model system in which we examined the effects of human surfactant protein A (SP-A) on the uptake of a common human pulmonary pathogen, Pseudomonas aeruginosa, by a human monocytic/macrophage cell line, THP-1 cells. We found that SP-A significantly increases uptake of the bacteria in a dose-dependent manner. Bacterial uptake was temperature-dependent, because an effect of SP-A on bacterial uptake was observed at 37°C and not at 4°C. The continued presence of SP-A during the period when the bacteria and THP-1 cells were co-incubated was necessary for enhanced uptake. Pre-incubation of the bacteria or THP-1 cells with SP-A, followed by washing, abolished the effect of SP-A on bacterial uptake. The effect of SP-A could be inhibited by high concentrations of mannose, but was not affected by the removal or addition of lipopolysaccharide (LPS). Finally, we observed that the SP-A-mediated increase in uptake of P. aeruginosa by THP-1 cells was optimal in a narrow (100 mM and 150 mM) range of NaCl concentrations. We conclude that SP-A enhances the THP-1 cell-mediated uptake of P. aeruginosa in a manner dependent on temperature, the concentration of SP-A, and the concentration of NaCl.

Abbreviations: analysis of variance, ANOVA; airway surface liquid, ASL; bronchoalveolar lavage, BAL; bovine serum albumin, BSA; cystic fibrosis, CF; cystic fibrosis transmembrane conductance regulator, CFTR; endotoxin unit, EU; fluorescence-activated cell sorter, FACS; fetal bovine serum, FBS; fluorescein isothiocyanate, FITC; forward scatter detector, FSC; lipopolysaccharide, LPS; mannose-binding protein, MBP; phosphate-buffered saline, PBS; phorbol 12-myristate 13-acetate, PMA; standard error, SE; secretory leukoproteinase 1, SLP1; surfactant protein A, SP-A; side scatter detector, SSC.




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