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Post-treatment effect of deferiprone in infected primary cells: Protein, RNA, and DNA of HIV-1.

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posted on 2016-05-18, 08:40 authored by Deepti Saxena, Michael Spino, Fernando Tricta, John Connelly, Bernadette M. Cracchiolo, Axel-Rainer Hanauske, Darlene D’Alliessi Gandolfi, Michael B. Mathews, Jonathan Karn, Bart Holland, Myung Hee Park, Tsafi Pe’ery, Paul E. Palumbo, Hartmut M. Hanauske-Abel

Cultures were infected with clinical isolate of HIV-1 on Day 0 as described [43]. Once self-sustaining infection was established by Day 7, cultures were treated with 200 μM deferiprone for the indicated duration. Controls were identically maintained without drug. Observation of virological parameters was extended to 10 days after cessation of medication. Viral load in untreated controls, under culture conditions consistently stabilizing within a narrow range at 106 copies/ml throughout month-long experiments (see Figs 1 and 5), are shown for the transit from on-drug to off-drug. Each p24 value is expressed relative to the respective p24 control on the day of each measurement. Results of the HIV-1 DNA determination are expressed according to the A450-based gradation of the assay, which defines A450 readings of <0.350 as ‘0 copies’, emphasized by the green arrow at the HIV-1 DNA axis, and increases stepwise to ‘20+ copies’ at A450 readings above 3.000 (Roche Amplicor HIV-1 DNA Test; see Materials and Methods). Triangles, viral p24; circles, viral RNA; squares, viral DNA; closed symbols, treatment period; open symbols, pre- and post-treatment periods; black asterisk, cessation of medication; red line segment, HIV-1 RNA decline off-drug; arrowheads, half of each culture replenished with fresh medium, drug, and primary cells; blue, control parameters.