Associations of full-length <i>KIR2DS4</i> gene with longitudinal viral load (VL) and CD4⁺ T-cell (CD4) count in 207 youth with chronic (seroprevalent) HIV-1 infection.

<p><b>Notes</b>: The overall <i>r</i>² = 0.039 and 0.033 in the multivariable models for VL and CD4 count, respectively. There is no clear interaction between full-length <i>KIR2DS4</i> (gene) and sex (<i>p</i> = 0.274).</p

Linear correlation between HIV-1-related outcomes and KIR2DS4 expression in the absence of antiretroviral therapy.

<p>Results are based on data from 23 subjects with full-length <i>KIR2DS4</i> gene (samples drawn from a single visit). <b>A</b>. Correlation of KIR2DS4 expression with plasma HIV-1 viral load (VL) at the time of sampling. <b>B</b>. Correlation with CD4⁺ T-cell (CD4) count (cells/µL) at the time of sampling. In each panel, solid and dotted lines correspond to the projected trend line and its 95% confidence intervals, respectively (by linear regression).</p

<i>KIR2DS4</i> gene expression and natural killer (NK) cell function in subjects with chronic HIV-1 infection.

<p><b>A</b>. Flow cytometry gating strategy for NK cells. <b>B</b>. Staining of membrane-bound KIR2DS4 on cells derived from subjects with full-length <i>KIR2DS4</i> gene (g+), to facilitate the sorting of two populations positive (p+) or negative (p−) for the gene product. <b>C</b>. Staining of membrane-bound KIR2DS4 on cells derived from subjects with truncated KIR2DS4 gene only, i.e., negative for full-length <i>KIR2DS4</i> gene and gene product (g−/p−). <b>D</b>. Percentage of NK cells expressing KIR2DS4 before and after stimulation with HLA-deficient target cells (K562 and 221). <b>E</b>. Distribution of NK cell subsets among 43 subjects with and without the full-length <i>KIR2DS4</i> genotype. <b>F–H</b>. Representative results for staining cell surface CD107a (lysosomal-associated membrane protein 1) and intracellular IFN-γ or MIP-1β before (red) and after (blue) stimulation with K562 cells. In <b>D</b> and <b>E</b>, the horizontal bars connected by a vertical line correspond to the median and interquartile range.</p

Characteristics of two study populations with longitudinal and cross-sectional data, respectively.

<p><b>Notes</b>: HIV-1 viral load (VL) in plasma (RNA copies/mL) and CD4⁺ T-cell (CD4) count in peripheral blood (cells/µL) are the two outcomes (see text). The two subgroups with existing PBMC samples share similar features at study entry (<i>p</i>>0.45 in all comparisons).</p

Assessment of polyfunctional profile of natural killer (NK) cells during chronic HIV-1 infection.

<p>Results are obtained after stimulation with K562 cells and subtraction of background staining. <b>A</b>. Functional properties of NK cells grouped by presence (+) and absence (−) of full-length <i>KIR2DS4</i> (g = gene) and membrane-bound KIR2DS4 receptor (p = product), as gauged by production of CD107a, IFN-γ and MIP-1β. <b>B</b>. Summary data for mono- and poly-functional NK cells. <b>C</b>. Distribution of polyfunctional NK cells. <b>D</b>. Distribution of NK cells NK cells producing MIP-1β alone. The g+/p+ and g+/p− cells are derived from individuals carrying full-length <i>KIR2DS4</i> gene, while the g−/p− NK cells are derived from individuals with truncated <i>KIR2DS4</i> gene only. In <b>C</b> and <b>D</b>, the horizontal bars connected by a vertical line correspond to the median and interquartile range.</p

Functional coordination within HIV-infected subject groups.

<p><b>A.</b> The extent of functional coordination within groups was assessed by calculating Spearman’s rank correlation coefficients across each pair of independently assessed functional assays. Differences between subject groups were evaluated using a Friedman ANOVA corrected for multiple comparisons using Dunn’s Test in Graphpad Prism. <b>B</b>. Prevalence of functional correlations by strength for each subject group. <b>C</b>. Correlation matrix for each pairwise combination of functions tested, in which strong positive correlations appear blue while inverse correlations appear red, for each subject group. Correlative relationships and significance were calculated and visualized using R, with unadjusted p values indicated to facilitate relative comparisons.</p

Antibody functionality can be predicted by subclass composition.

<p><b>A</b>. Correlative relationships between levels of total gp120-specific IgG or gp120-specific antibodies of each subclass to antibody function were assessed by determination of Spearman’s rank correlation coefficients between activity and antibody MFI within each subject group. <b>B</b>. Correlative relationships between <i>relative</i> levels of gp120-specific antibodies of each subclass (i.e., MFI of subclass/MFI of total IgG) were assessed for each functional activity, over all subjects. Positive associations are noted in blue and inverse associations in red, with the magnitude of correlation depicted by intensity. Correlative relationships and significance were calculated and visualized using R, with unadjusted p values indicated to facilitate relative comparisons. <b>C</b>. The magnitude and direction of the contribution of SF162-specific antibody subclass assessments to cross-validated predictive models of polyfunctional activity.</p

Levels and IgG subclass composition differentiate HIV+ subject groups.

<p><b>A.</b> Titer (Mean Fluorescent Intensity, MFI) of gp120-specific IgG present in each subject group. <b>B</b>. The percent of subjects in each group positive for gp120-specific responses of each IgG subclass. <b>C.</b> Spearman correlation matrix between subclass responses across subjects. Strength and significance, as calculated in Graphpad Prism, are represented as color intensity and size, respectively. <b>D.</b> The levels of gp120-specific responses observed across cohort groups for each IgG subclass. Differences between groups were assessed by Kruskal-Wallis ANOVA and corrected for multiple comparisons using Dunn’s test in Graphpad Prism.</p

Clinical characteristics of HCV-infected individuals.

<p>*p.i. =  post infection; Risk factor for HCV acquisition; GT =  HCV genotype; ND = not determined. # normal range of ALT = 7–56 IU/ml, AST = 5–40 IU/ml.</p

Immunophenotypic profiles of blood and liver-resident NK cell subsets.

<p>Immunophenotyping of liver and blood NK cell subsets in groups of HCV-infected and –uninfected individuals was performed by flow cytometry, and both tissue-specific differences (A) and disease-specific differences (B) were analyzed. Data is presented as a heatmap, with values displayed as relative within each row. Statistical significance was accepted at p<0.05 and is indicated by * (p<0.05), ** (p<0.01), *** (p<0.001) and **** (p<0.0001).</p