Ontogeny of Salivary Human Beta-Defensin-1 and Lactoferrin

Antimicrobial peptides (AMPs), an important component of innate immunity in the oral cavity, provide a first line of host defense against bacterial, viral and fungal challenges. These peptides may play a vital role in saliva, where the oral microbiota present constant and varied challenges to the oral mucosa. Objective: To explore the association of salivary AMPs, hBD-1 and lactoferrin, with age in a cross-sectional study.  Methods: A 4 ml whole, unstimulated saliva sample was obtained from each of 183 healthy subjects (age 8 to 78). Periodontal measures of the 6 Ramjford teeth were measured on adult subjects. Decayed/Missing/Filled Teeth (DMFT) and gingival indices were obtained on all subjects. Levels of hBD-1 were measured by Sandwhich ELISA following the method of Ghosh et al (2007) using antibody pairs from Peprotech (NJ, USA). Recombinant hBD-1 peptide (Peprotech, NJ) was used as standard.  Levels of lactoferrin were measured using an ELISA kit from Hycult Biotech (Plymouth Meeting, PA) following the vendor's instructions. Concentrations of the AMPs were normalized with total salivary protein. Spearman rank-order correlation coefficient was used to examine AMPs and between AMPs with age. Results: Median levels of hBD-1 and lactoferrin (ng/mg protein) standardized by total protein were 195.0 (32.0 – 965.6) and 2619.4 (88.8 – 14,641.4), respectively. We found no direct or inverse coreelation between hBD-1 and lactoferrin with age, and only modestly for hBD-1 with periodontal measure. Conclusions: Unlike other AMPs, such as SLPI ,calprotectin, hBD-2 and LL37 [Shugars et al, Gerontology, 2001 47(5):246-53;  VanYperen et al, Abstract# 860 and  Demko et al, Abstract# 863, IADR 2011, San Diego, CA ] that were found to correlate with age, hBD-1 and lactoferrin did not. Supported by  Supported by NIH/NIDCR RO1DE16334 (AW)