Fig. 1
Viral load and immune cell subsets in chronically infected HIVSF162 humanized mice suppressed or not for osteopontin gene and protein expression. a Viral loads measured by Taqman qRT-PCR assay with a sensitivity of detection of 1 copy/ml, varied among individual female mice (symbols) (blue, HIV-OPN-, n = 8; red, HIV-OPN+, n = 3) in each group (simple linear regressions, no significance differences in slope, F = 1.471. DFn = 2, DFd = 44, P = 0.2408). b Similarly, no significant differences in viral load between the HIV OPN+ and HIV OPN- groups before or after aptamer treatment were found (F = 1.329. DFn = 2, DFd = 13, P = 0.2983). c–e Percentage of white blood cells in plasma of HIV OPN+ (red) and HIV OPN- (blue) female mice before HIV infection and 85 days after aptamer treatment. c Absolute percentage of human CD45+ cells (HIV OPN+, red, n = 5, start; n = 2 end; HIV OPN- (blue, n = 7, start; n = 7 end; F = 3.800. DFn = 1, DFd = 17, P = 0.0680). d percentage of hCD4 in the total hCD3+ T cell population (HIV OPN+, red, n = 5, start; n = 2 end; F = 0.3051. DFn = 1, DFd = 17, P = 0.5879); HIV OPN+ (blue, n = 8, start; n = 6 end). e Percentage of hCD8+ in the total hCD3+ T cell population (HIV OPN+, red, n = 5, start; n = 2 end; HIV OPN- (blue, n = 7, start; n = 7 end; F = 1.489. DFn = 1, DFd = 17, P = 0.2390). f RNAscope was used to detect HIV RNA in spleen and brain of HIV-infected mice. Panels on left (top to bottom): negative control for DapB, positive controls (green = PPIB DNA, red = POLR2A, RNA), HIV RNA in the spleen. Middle panel: HIV RNA in brain parenchyma with enlarged panels on the right showing localization to cytoplasm of cells. g Representative sections of gut (small intestines) stained with hematoxylin/eosin (top panels) or immunostained for OPN protein (lower panels). h Quantification of OPN protein immunoreactivity from female mice gut (each 10-μm section has the entire gut on the slide) subjected to neuroimaging (top graphic representation: n = 3 (labeled m43, etc.), 9–12 images quantified per mouse, each dot represents an image). Grouped analyses, ordinary two-way ANOVA, multiple comparisons, Tukey’s, 95% CI, P < .0001, ****. There was an interaction between HIV and OPN (two-way ANOVA, F = 6.56. DFn = 1, DFd = 104, P = 0.0118) and independently with HIV that accounted for 15.02% of the total variance (two-way ANOVA, F = 21.96. DFn = 1, DFd = 104, P < 0.0001) and with OPN that was responsible for 9.36% of the total variance (two-way ANOVA, F = 13.69. DFn = 1, DFd = 104, P = 0.0003). i Quantification of OPN mRNA levels in brain tissue of female and all (female and male mice) in each group. Ordinary one-way ANOVA, multiple comparisons, Tukey’s, 95% CI; [mean, std]: females: [n = 4, Buffer OPN+ 10.14, 12.19], [n = 3, Buffer OPN- 0.504, 0.512], [n = 5, HIV OPN+ 4.41, 5.71], [n = 6, HIV OPN- 2.86, 5.9]; all: [n = 7, Buffer OPN+ 12.68, 16.13], [n = 5, Buffer OPN- 0.363, 0.424], [n = 8, HIV OPN+ 6.14, 9.37], [n = 9, HIV OPN- 1.89, 4.89]