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. 2016 Mar 1;29(2):112–127. doi: 10.1089/vim.2015.0105

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Copyright 2016, Mary Ann Liebert, Inc.

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FIG. 3. — Mean frequencies of CD4 T cells (a), CD8 T cells (b), Treg cells (c), and NK cells (d) among lymphocytes (A) and mean numbers per tissue (B). MNCs were mock stimulated for 17 h (Brefeldin A was added for the last 5 h) and then subjected to intracellular staining and flow cytometry for collecting the frequency and number data of CD4 T cells (a), CD8 T cells (b), and NK cells (d) and served as background controls for the data of virus-specific IFN-γ-producing cell frequencies (Fig. 4). MNCs were stained freshly for Treg cells (c) without in vitro stimulation. The absolute number of CD4 and CD8 T cells, Treg cells, and NK cells per tissue was calculated based on their frequencies and the total number of MNCs isolated from each tissue. The left figures show the prechallenge data at PID 28 and those in the right show the postchallenge data at PCD 7. Data are presented as mean frequency/number ± standard error of the mean (n = 3 or 5). The asterisk “*” indicates significant difference in frequencies/numbers among groups for the same cell type and tissue; different number of “*” indicates significant differences among groups (Kruskal–Wallis test, p < 0.05). IFN-γ, interferon-gamma; NK, natural killer; Treg, regulatory T.

FIG. 3. — Mean frequencies of CD4 T cells (a), CD8 T cells (b), Treg cells (c), and NK cells (d) among lymphocytes (A) and mean numbers per tissue (B). MNCs were mock stimulated for 17 h (Brefeldin A was added for the last 5 h) and then subjected to intracellular staining and flow cytometry for collecting the frequency and number data of CD4 T cells (a), CD8 T cells (b), and NK cells (d) and served as background controls for the data of virus-specific IFN-γ-producing cell frequencies (Fig. 4). MNCs were stained freshly for Treg cells (c) without in vitro stimulation. The absolute number of CD4 and CD8 T cells, Treg cells, and NK cells per tissue was calculated based on their frequencies and the total number of MNCs isolated from each tissue. The left figures show the prechallenge data at PID 28 and those in the right show the postchallenge data at PCD 7. Data are presented as mean frequency/number ± standard error of the mean (n = 3 or 5). The asterisk “*” indicates significant difference in frequencies/numbers among groups for the same cell type and tissue; different number of “*” indicates significant differences among groups (Kruskal–Wallis test, p < 0.05). IFN-γ, interferon-gamma; NK, natural killer; Treg, regulatory T.