Dynamics of X4 WT and <i>vpr</i>-deficient HIV-1 infection in humanized mice.

<p>(A and B) CXCR4 expression on CD4⁺ T cell subsets in human and humanized mouse. Human CD4⁺ T cells isolated from the spleen of humanized mice (A, n = 8) and the PB of HIV seronegative humans (B, n = 6) were classified into Tn, Tm, and Treg as described in the legend of <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003812#ppat-1003812-g001" target="_blank">Figure 1</a>. Representative dot plots and histograms are shown on the left, and the percentages of CXCR4⁺ cells in each subset are shown on the right. In the left panels, the numbers in each histogram indicate the positivity. (C) Viral load in infected humanized mice. The amounts of viral RNA in the plasma of X4 WT HIV-1-infected mice (n = 13) and X4 <i>vpr</i>-deficient HIV-1-infected mice (n = 11) were routinely quantified. The horizontal broken line indicates the detection limit of the assay (1,600 copies/ml). (D) Longitudinal analyses of the dynamics of human CD4⁺ T cell subsets in the PB of infected humanized mice. The numbers of total CD4⁺ T cells, Tns, Tms, and Tregs in the PB of WT HIV-1-infected mice (n = 9), <i>vpr</i>-deficient HIV-1-infected mice (n = 9), and mock-infected mice (n = 8) were routinely quantified by flow cytometry and hematocytometry. (E and F) Cytopathic effect of WT and <i>vpr</i>-deficient HIV-1 in the spleen of humanized mice. The percentages of total CD4⁺ T cells, Tns, Tms, and Tregs in the splenic MNCs of WT HIV-1-infected mice (n = 8), <i>vpr</i>-deficient HIV-1-infected mice (n = 8), and mock-infected mice (n = 8) at 21 dpi were routinely quantified by flow cytometry. Representative dot plots (E) and summarized results (F) are shown, respectively. In panel E, the numbers on the right of the dot plots indicate the percentage of the cells in each quadrant. (G) The level of immune activation in infected humanized mice. The MFI of CD38 in memory CD8⁺ T cells in the spleen of WT HIV-1-infected mice (n = 5), <i>vpr</i>-deficient HIV-1-infected mice (n = 5), and mock-infected mice (n = 5) at 21 dpi was analyzed by flow cytometry. Representative histograms are shown on the right panel, and the numbers in the histogram indicate the MFI values. NS, no statistical significance. Data represent mean ± SEM.