The effect of ciprofloxacin, a quinolone antibiotic widely used to treat opportunistic bacterial infections in AIDS patients, was examined in the context of reactivation of latent HIV-1 in chronically infected promonocytic U1 cells. Ciprofloxacin inhibited, in a dose-dependent manner, HIV-1 expression in U1 cells activated with phorbol 12-myristate 13-acetate (PMA). The inhibitory effect of ciprofloxacin was associated with a reduction in the production of tumor necrosis factor alpha, inhibition of activation of transcriptional factor NF-kappaB, and HIV LTR-driven gene expression. Furthermore, ciprofloxacin inhibited TNF-alpha-induced HIV expression in U1 cells. The concentrations of ciprofloxacin that inhibited HIV production are readily achievable in vivo.