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. 2016 Aug;186(8):2068–2087. doi: 10.1016/j.ajpath.2016.03.019

Figure 4.

Figure 4

Changes in spleen monocyte cells during SIV infection. A: Single-cell suspensions were generated from spleens isolated from uninfected and SIV-infected macaques during the acute (4, 7, and 10 dpi), asymptomatic (14 and 21 dpi), and chronic (56 and 84 dpi) phases of infection. Cell suspensions were analyzed by flow cytometry to determine the frequency of monocytes (gray bars) and T lymphocytes (white bars) in the spleen leukocyte population. B: Spleen cell suspensions were analyzed by flow cytometry to determine the frequency of monocytes that expressed cell surface CD16 in uninfected animals and at 7, 21, 56, and 84 dpi. C: RNA was isolated from the spleen from uninfected animals and at 4, 7, 14, 21, and 56 dpi and CD14 gene expression analyses performed by NanoString barcoding technology. The relative count of CD14 was determined on normalizing to four housekeeping genes, and the fold-change in the SIV-infected animals relative to the uninfected controls, which was set to 1, determined. D: Whole blood was collected and immunostained with fluorochrome-coupled anti-CD14 to determine the frequency of peripheral blood CD14 monocytes by flow cytometry in uninfected animals and at 4, 7, 10, 14, 21, 56, and 84 dpi. The acute (light shading), asymptomatic, and chronic (dark shading) phases of SIV infection are depicted. Solid lines connecting data points denote cumulative and longitudinal blood sampling of the same animals at every time point after inoculation. Dashed lines connecting data points indicate cross-sectional study of independent groups of animals after euthanasia at each time point. A, B, and D: Statistical significance was determined by Mann-Whitney test. Statistical significance was determined by Kruskal-Wallis analysis of variance test followed by Dunn's multiple comparison analysis. C: Vertical lines indicate a significant difference between the indicated dpi and uninfected animals, as determined by the Dunn's multiple comparison analysis. Data are expressed as means ± SD. ∗P ≤ 0.05, T cells versus the preceding day after inoculation; †P ≤ 0.05 for monocytes versus the preceding day after inoculation; ‡P ≤ 0.05 relative to the preceding day after inoculation; §P ≤ 0.05 for infected animals at all time points relative to uninfected macaques.