Human beta-defensin-2, and -3 (hBD-2, -3) are small inducible antimicrobial peptides involved in host defense. Actinobacillus actinomycetemcomitans, a gram-negative facultative anaerobe, is frequently associated with oral disease in humans. A. actinomycetemcomitans, strain JP2, was examined for its ability to modulate hBD-2 and -3 gene expression in normal human oral epithelial cells (NHOECs) and in OKF6/Tert cells, an immortalized cell line derived from human oral epithelial cells. Stimulation of both cell types by live bacteria, at a minimal bacteria/cell ratio of 500 : 1, resulted in increased hBD-3 gene expression. This was not evinced for hBD-2 in either cell type with live bacteria, even at bacteria/cell ratios exceeding 500 : 1. The increased hBD-3 gene expression was dependent upon viable bacteria, and not their lipopolysaccharides (LPS), since heat-killed A. actinomycetemcomitans did not induce hBD-3 transcript expression. The overall similarity between results obtained in OKF6/Tert cells and NHOECs suggest that the OKF6/Tert cell line may be a useful tool in the study of beta-defensin expression in oral epithelium.