A549 Lung Epithelial Cells Grown as Three-Dimensional Aggregates: Alternative Tissue Culture Model forPseudomonas aeruginosaPathogenesis
| Autor: | Duane L. Pierson, Mark S. F. Clarke, Kent L. Buchanan, Alexander J. Carterson, C. M. Ott, K. Höner zu Bentrup, Michael J. Schurr, Charles R. Vanderburg, Cheryl A. Nickerson |
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| Rok vydání: | 2005 |
| Předmět: |
Collagen Type IV
Immunology Cell Culture Techniques Mucin 5AC Biology Models Biological Microbiology Tissue culture Bioreactors Antigen Antigens Neoplasm medicine Humans Pseudomonas Infections Antigens Lung MUC1 Glycoproteins A549 cell Tight junction Tumor Necrosis Factor-alpha Interleukins Mucin-1 Mucins Microcarrier Epithelial Cells Bacterial Infections respiratory system Molecular biology Epithelium respiratory tract diseases Infectious Diseases medicine.anatomical_structure Cell culture Pseudomonas aeruginosa Parasitology Laminin Biomarkers |
| Zdroj: | Infection and Immunity. 73:1129-1140 |
| ISSN: | 1098-5522 0019-9567 |
| DOI: | 10.1128/iai.73.2.1129-1140.2005 |
| Popis: | A three-dimensional (3-D) lung aggregate model was developed from A549 human lung epithelial cells by using a rotating-wall vessel bioreactor to study the interactions betweenPseudomonas aeruginosaand lung epithelial cells. The suitability of the 3-D aggregates as an infection model was examined by immunohistochemistry, adherence and invasion assays, scanning electron microscopy, and cytokine and mucoglycoprotein production. Immunohistochemical characterization of the 3-D A549 aggregates showed increased expression of epithelial cell-specific markers and decreased expression of cancer-specific markers compared to their monolayer counterparts. Immunohistochemistry of junctional markers on A549 3-D cells revealed that these cells formed tight junctions and polarity, in contrast to the cells grown as monolayers. Additionally, the 3-D aggregates stained positively for the production of mucoglycoprotein while the monolayers showed no indication of staining. Moreover, mucin-specific antibodies to MUC1 and MUC5A bound with greater affinity to 3-D aggregates than to the monolayers.P. aeruginosaattached to and penetrated A549 monolayers significantly more than the same cells grown as 3-D aggregates. Scanning electron microscopy of A549 cells grown as monolayers and 3-D aggregates infected withP. aeruginosashowed that monolayers detached from the surface of the culture plate postinfection, in contrast to the 3-D aggregates, which remained attached to the microcarrier beads. In response to infection, proinflammatory cytokine levels were elevated for the 3-D A549 aggregates compared to monolayer controls. These findings suggest that A549 lung cells grown as 3-D aggregates may represent a more physiologically relevant model to examine the interactions betweenP. aeruginosaand the lung epithelium during infection. |
| Databáze: | OpenAIRE |
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