Flow cytometric analyses of the expression of human gp91^phox by high side scatter human CD45⁺ cells in the bone marrow of NOD/SCID mice that received transplants of human CD34⁺PBSCs.

Shown are representative dot plots of analyses of gp91^phoxexpression in human (CD45⁺) cells engrafted in chimeric bone marrow from NOD/SCID mice that underwent transplantation with 4-day cultured but nontransduced X-CGD CD34⁺PBSCs (A,C), lentivector–gp91^phox-transduced X-CGD CD34⁺PBSCs (B); MFGS–gp91^phox-transduced X-CGD CD34⁺PBSCs (D), and 4-day cultured but nontransduced normal CD34⁺PBSCs (E). Side scatter is plotted on the vertical axis, and labeling with antihuman gp91^phox is plotted on the horizontal axis. Analyses are gated to include only those cells that label positive for the CD45 antigen, as depicted in Figure 7. Boxed areas represent the events that fall into the gp91^phox-positive region. Labeling and analyses in the different panels were performed on different days and resulted in a variability of dot plots. Of note is that the PCR-measured copy number of vector insert per engrafted human cell in each mouse correlates proportionately with the percentage of human cells expressing transgene by flow cytometry in each mouse, though silencing would alter the apparent ratio.