BACKGROUND The objectives of this study were to determine 1) oral hBD2 expression in HIV-infected subjects compared to non-HIV controls, 2) the expression of oral hBD2 in HIV-infected subjects with ART compared with those without ART, and 3) factors associated with the expression of oral hBD2. METHODS Oral examination and punched biopsy on buccal mucosa were performed in HIV-infected subjects with and without ART, and non-HIV individuals. The expression of hBD2 mRNA was determined by quantitative real-time PCR. Saliva samples of both un-stimulated and stimulated saliva were collected and analyzed for hBD2 levels using ELISA. Student’s t-test and nonparametric multi-way ANOVA test were used for comparison of measurements between or among groups. RESULTS One hundred and fifty-seven HIV-infected subjects were enrolled; 99 on ART (age range 23–57 yr, mean 39 yr), 58 not on ART (age range 20–59 yr, mean 34 yr), and 50 non-HIV controls (age range 19–59 yr, mean 36 yr). The most common ART regimen was 2 NRTIs+1 NNRTI. Salivary levels of hBD2 were significantly increased in HIV infection (p< 0.001). The levels of hBD2 in stimulated saliva were also found to be significantly different between HIV-infected subjects who were and were not on ART (p< 0.001). No significant difference was observed with the expression of hBD2 mRNA. CONCLUSION Oral innate immunity is affected by HIV infection and use of ART. Salivary hBD2 levels may be the useful biomarkers to monitor those on long-term ART who are at risk of developing oral infections and malignant transformation.