RNA genome of HIV-1 reverse transcribed in cells of Schistosoma mansoni and lentiviral cDNA integrated into the schistosome genome.

Panel A. Quantitation of negative-strand, strong-stop HIV-1 cDNA in genomic DNA of schistosomula 24 hours after exposure to intact or heat-inactivated virions. Panel B. Quantitation of the late, positive-strand HIV-1 cDNA in schistosomula at 24 and 48 hours after incubation with virions. Panel C. Schematic representation of the nested two-step, quantitative retrotransposon-anchored PCR (qRAP) for relative quantitation of HIV-1 provirus integrated into the schistosome genome. Products of the first reaction using retrotransposon-specific primers were subjected to secondary PCR using provirus-specific primers. Panel D. Detection of HIV-1 provirus integrated into the schistosome genome using the primer set no. 1 specific for retrotransposons SR1 and SR2. Panel E. Detection of HIV-1 provirus detected with primer set no. 2, specific for fugitive, SMα, and Boudicca. Statistical analysis: Student’s t-test; *, **—P ≤ 0.05, P ≤ 0.01 (active vs. heat-inactivated virions). The experiments were repeated ≥ three times.