Assay of nascent viral DNA synthesis in monocytes with or without SLPI treatment. Adherent monocytes were infected with HIV-1Ba-L for 2.5 hours, washed extensively, trypsinized to remove loosely bound virus particles, and then incubated for 18 hours. Cellular lysates were prepared and assayed for the presence of viral DNA using a nested-PCR–based assay. A 730-bp HIV-1–specific PCR product was detected in virus-infected cells (lane 2), but not in uninfected cells (lane 1). In the presence of 5 μg/mL SLPI, no virus-specific PCR product was generated (lane 3), indicating SLPI-mediated inhibition of viral DNA formation. Equivalent amounts of cellular DNA per sample were analyzed as indicated by PCR amplification using β-globin–specific primers (data not shown).
