Measuring Antimicrobial Peptide Activity on Epithelial Surfaces in Cell Culture

• DOI: 10.1007/978-1-60761-594-1_23
• Publication Date: 2010-01-21T11:16:14Z
• Authors: Gill Diamond; Sunghan Yim; Isaura Rigo; Laura McMahon
• Keywords: 0301 basic medicine; Cell Culture Techniques; Gingiva; Gene Expression; Bronchi; Epithelial Cells; Fibroblasts; Bordetella bronchiseptica; Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents; Cell Line; 03 medical and health sciences; Cathelicidins; Pseudomonas aeruginosa; Animals; Humans; Vitamin D; Cells, Cultured; Antimicrobial Cationic Peptides

To more accurately assess the activity and role of epithelial cell-derived antimicrobial peptides in their native settings, it is essential to perform assays at the surfaces under relevant conditions. In order to carry this out, we utilize three-dimensional cultures of airway and gingival epithelium, which are grown at an air-liquid interface. Under these conditions, the cultures can be subjected to challenge with a variety of factors known to cause an increase in antimicrobial peptide gene expression. The functional relevance of this induction can then be assessed by quantifying antibacterial activity either directly on the surface of the cells or using the fluid secreted onto the apical surface of the cultures. The relative contribution of the peptides can also be measured by pre-incubation of the secreted fluid with specific inhibitory antibodies. Thus, a relatively inexpensive in vitro model can be used to evaluate the role of antimicrobial peptides in mucosal epithelium.