Dynamics of human CD4⁺ T cell subsets in humanized mice infected with R5 WT and <i>vpr</i>-deficient HIV-1.

<p>(A and B) 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 (CD45⁺ CD3⁺ CD4⁺ cells), Tns (CD45⁺ CD3⁺ CD4⁺ CD45RA⁺ FOXP3⁻ cells), Tms (CD45⁺ CD3⁺ CD4⁺ CD45RA⁻ FOXP3⁻ cells), and Tregs (CD45⁺ CD3⁺ CD4⁺ CD45RA⁻ FOXP3⁺ cells) in the PB of R5 WT HIV-1-infected mice (n = 8), R5 <i>vpr</i>-deficient HIV-1-infected mice (n = 8), and mock-infected mice (n = 12) were routinely quantified by flow cytometry and hematocytometry. Summarized results (A) and representative dot plots at 21 dpi (B) are shown, respectively. In panel B, the numbers in the histogram indicate the percentage of CD4⁺ cells in CD45⁺ cells, and the numbers under the dot plots indicate the percentage of the cells in each quadrant. (C and D) 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 (7 dpi, n = 19; 21 dpi, n = 8), <i>vpr</i>-deficient HIV-1-infected mice (7 dpi, n = 10; 21 dpi, n = 7), and mock-infected mice (n = 12) were routinely quantified by flow cytometry. Representative dot plots (C) and summarized results (D) are shown, respectively. In panel C, the numbers under the dot plots indicate the percentage of the cells in each quadrant. (E) The level of CCR5-expressing CD4⁺ T cells in infected humanized mice. The percentage of CCR5⁺ cells in the splenic CD4⁺ T cells of WT HIV-1-infected mice (7 dpi, n = 8; 21 dpi, n = 6), <i>vpr</i>-deficient HIV-1-infected mice (7 dpi, n = 8; 21 dpi, n = 6), and mock-infected mice (n = 8) was analyzed by flow cytometry. (F) The level of immune activation in infected humanized mice. The MFI of CD38 in memory CD8⁺ T cells (CD45⁺ CD3⁺ CD8⁺ CD45RA⁻ 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. Statistical difference was determined by Welch's <i>t</i> test, and statistically significant differences (<i>P</i><0.05) are shown as follows: mock versus WT HIV-1, black asterisk; mock versus HIV-1<i>Δvpr</i>, blue asterisk; and WT HIV-1 versus HIV-1<i>Δvpr</i>, red asterisk. NS, no statistical significance. Data represent mean ± SEM.