Measuring antimicrobial peptide activity on epithelial surfaces in cell culture.
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.
Diamond
Gill
G
Department of Oral Biology, UMDNJ-New Jersey Dental School, Newark, NJ, USA.
Yim
Sunghan
S
Rigo
Isaura
I
McMahon
Laura
L
D016976:Aggregatibacter actinomycetemcomitans / Q000187:drug effects; D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D016950:Bordetella bronchiseptica / Q000187:drug effects; D001980:Bronchi / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D005881:Gingiva / Q000166:cytology; D006801:Humans; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D014807:Vitamin D / Q000378:metabolism; D054804:Cathelicidins
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D014807:Vitamin D; D054804:Cathelicidins