CD4 is expressed on a heterogeneous subset of hematopoietic progenitors, which persistently harbor CXCR4 and CCR5-tropic HIV proviral genomes in vivo
Fig 2
Near-full-length HIV provirus in HSPCs preparations is unlikely to be from T cell contamination.
A. Flow cytometric analysis of HSPCs from three donors that harbored near full-length genomes. Numbers indicate the frequency of events falling into each quadrant. CD133 or CD34 were assessed for Sort 1 or Sort 2 respectively. B. Summary table showing data for Fisher’s exact test, which compares calculated frequencies of provirus assuming all provirus originates from T cells. PCRs were performed at limiting dilution. Significantly discrepant frequencies in sorted and flow-through samples indicate T cells are an unlikely source of provirus in sorted samples. Conservative estimate compares the top of the 95% confidence interval for the calculated infection rate for flow-through CD3+ T cells with the bottom of the 95% confidence interval for the calculated infection rate for CD3+ T cells in the sorted HSPCs. C. Agarose gel analysis of rearranged T cell receptor gene PCR. 50, 100 or 200 cell equivalents of DNA from the first round PCR reaction that yielded each provirus were added to each reaction. For “T cells”, 200 cell equivalents of DNA from purified CD4+ T cells were added to the reaction. The expected size of the amplicons is 250–300 bp. Numbers to the left of each gel indicate the location of molecular weight markers.