High VF frequencies correlate with the efficiency of immature DC viral transfer.
2012-06-07T02:32:26Z (GMT) by
<p>(<b>A</b>) Schematic of the DC-CD4 T cell transfer assay. DC are infected with either HIV<sup>WT</sup> (high VF frequency) or HIV<sup>-NEF-ve</sup> (low VF frequency) pseudotyped with the VSVg glycoprotein, to ensure equal infection frequencies. After 4 days, DC infections are normalized to 5% with uninfected DC. Normalized populations are serially diluted below 1 infected DC per co-culture. 4 days post co-culture, CD4 T cell infections are resolved by staining cells for of HIV capsid and resolution by flow cytometry. (<b>B</b>) Flow cytometry detection of HIV p24 within CD4 T cell recipients when input infected DC are limiting (upper panel) versus (<b>C</b>) when input infected CD4 T cells are limiting (lower panel). Approximate infected cell number input into co-cultures is indicated at “Approx. Input*” on the X-axis. CD4 T cell infection frequencies are detected by the accumulation of a high HIV p24 population as indicated by the square gate in each dot-plot. Statistical difference is presented in upper HIV WT panels, and is calculated from data acquired from the same assay performed in triplicate. CD4 T cell infection frequency versus infected donor input is further summarized in right panels for two representative donors. Standard deviations represent co-cultures in triplicate. Data is representative of <i>n</i> = 8 independent donors.</p>