Whole blood monocyte proinflammatory cytokine production in HIVneg and HIV+ donors or SIVneg and SIV+ mangabeys following exposure to <i>M</i>. <i>bovis</i> BCG.

<p>(A) Representative flow cytometry plots and gating strategy of cytokine-producing monocytes, which were first defined as being live and CD3 negative (not shown), CD14+, and producing TNF-α, IL-12, or both following 6h exposure to BCG. (B-G) Percentage of TNF-α+ (B, E), IL-12+ (C, F), or double-positive (TNF-α+ and IL-12+) monocytes (D, G) in HIVneg (unfilled circles) and ART-naive HIV+ (filled circles) humans or SIVneg (unfilled squares) and SIV+ (filled squares) mangabeys following ex vivo BCG exposure. Lines represent the medians for each group (*p<0.05, Mann-Whitney).</p

Nonpathogenic SIV and Pathogenic HIV Infections Associate with Disparate Innate Cytokine Signatures in Response to <i>Mycobacterium bovis</i> BCG

<div><p>Infections with mycobacteria, including <i>Mycobacterium tuberculosis</i> (Mtb) and <i>Mycobacterium bovis (M</i>. <i>bovis)</i> BCG, are a leading cause of morbidity and mortality for HIV-infected persons. In contrast to HIV, nonpathogenic SIV infections of sooty mangabeys are characterized by a lack of clinical disease including an absence of opportunistic infections. The goal of this study was to identify innate immune responses to <i>M</i>. <i>bovis</i> BCG maintained during nonpathogenic lentiviral infections through a comparison of functional responses during pathogenic HIV or nonpathogenic SIV infections. Monocytes were evaluated for their ability to express key anti-mycobacterial cytokines TNF-α and IL-12 following a six-hour <i>ex vivo</i> BCG exposure. While HIV-infection was associated with a decreased percentage of IL-12-producing monocytes, nonpathogenic SIV-infection was associated with an increased percentage of monocytes producing both cytokines. Gene expression analysis of PBMC following <i>ex vivo</i> BCG exposure identified differential expression of NK cell-related genes and several cytokines, including IFN-γ and IL-23, between HIV-infected and control subjects. In contrast, SIV-infected and uninfected-control mangabeys exhibited no significant differences in gene expression after BCG exposure. Finally, differential gene expression patterns were identified between species, with mangabeys exhibiting lower IL-6 and higher IL-17 in response to BCG when compared to humans. Overall, this comparison of immune responses to <i>M</i>. <i>bovis</i> BCG identified unique immune signatures (involving cytokines IL-12, TNF-α, IL-23, IL-17, and IL-6) that are altered during HIV, but maintained or increased during nonpathogenic SIV infections. These unique cytokine and transcriptome signatures provide insight into the differential immune responses to Mycobacteria during pathogenic HIV-infection that may be associated with an increased incidence of mycobacterial co-infections.</p></div

Genes associated with HIV-status-specific effects following BCG exposure ex vivo.

<p>Genes associated with HIV-status-specific effects following BCG exposure ex vivo.</p

Whole PBMC cytokine gene regulation in HIVneg or HIV+ donors following BCG exposure. Plots demonstrate regulation of key proinflammatory.

<p>(A), Th1 (B), Th17 (C) and immunoregulatory genes (D) following 4h BCG exposure. Log2 copy number of RNA in an unstimulated control (U) or matched BCG-stimulated (B) samples in either humans (circles) or mangabeys (squares). The lower bound of the gray shaded portion of the graph represents the mean of the negative controls, and the upper bound represents two standard deviations above the mean, which ultimately determined the cutoff for expression. The most highly up- and down-regulated genes in humans or mangabeys are displayed via a heat map (E). FC: fold change; BD: below detection.</p

Model of differential cellular responses during the innate immune response to BCG during pathogenic HIV or non-pathogenic SIV infection.

<p>Cytokines that are differentially expressed between HIVneg and HIV+ humans (A) or SIVneg and SIV+ mangabeys (B) are presented without boxes and are in bold type font. Arrow(s) represent the up or down modulation of these immune modulators in response to BCG, and fold change (FC) is presented as numeric values. Cytokines expressed differently between human subjects and mangabeys are boxed (B).</p

Selected genes that demonstrate HIV status-specific effects of BCG exposure.

<p>Fold change in the expression of 199 immunologic genes in response to 4h BCG exposure of PBMC from HIVneg (circles) or ART-naïve HIV+ (squares) was assessed via Nanostring gene expression analysis. Twelve genes remained significant following correction for multiple comparisons at an FDR threshold <10% (listed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0158149#pone.0158149.t001" target="_blank">Table 1</a>). Following the identification of these genes, further validation of the differences in response between HIVneg and HIV+ subjects was identified via a paired t-test: PRF1 (HIVneg p = 0.0045, HIV+ p = n.s), and NKp46 (HIVneg p = 0.0008, HIV+ p = n.s) perform cellular effector functions, and CCR5 (HIVneg p = 0.0029, HIV+ p = n.s) is an HIV coreceptor.</p

HIV infection is associated with a deficiency in IFNg mRNA and protein upregulation in response to M. bovis BCG.

<p>mRNA expression of IFN-γ in response to 4h of BCG exposure from HIVneg or ART-naïve HIV+ PBMC was measured via Nanostring analysis (left panel; HIVneg p<0.0001, HIV+ p = 0.02). IFN-γ protein was measured in supernatants from matched PBMC stimulations at an 18h time point (right panel; HIVneg-unstim vs HIVneg BCG p = 0.0001; HIV+ unstim vs HIV+ BCG p = 0.004; HIVneg-BCG vs HIV+ BCG p<0.0001; HIVneg unstim vs. HIV+ BCG n.s.). Statistical evaluation was performed via a two-tailed t-test on log-transformed data.</p

Gene expression analysis of SIVneg and SIV+ mangabey PBMC following 4 hour BCG exposure.

<p>Semi-supervised hierarchical clustering via Principal Component Analysis (PCA) of 199 immunological gene targets (chosen <i>a priori</i>) was performed on RNA from whole PBMC following 4h BCG exposure and controls (without BCG), in SIV+ or SIVneg mangabeys via PCA (A). PC-1 (x-axis) represents the component responsible for the largest variation in the data set and explains 30% of the variation, while PC-2 (y-axis) represents the component contributing the second highest degree of variability in the dataset of 23%. Fold change in expression of these genes in SIVneg (x-axis) or SIV+ (y-axis) subjects after BCG exposure is depicted via scatterplot (B). No genes were found to diverge significantly from the trendline. A black outline denotes the genes that displayed statistically significant differences between HIVneg and HIV+ donors, for comparison.</p

Genetic architecture of human plasma lipidome and its link to cardiovascular disease

Abstract Understanding genetic architecture of plasma lipidome could provide better insights into lipid metabolism and its link to cardiovascular diseases (CVDs). Here, we perform genome-wide association analyses of 141 lipid species (n = 2,181 individuals), followed by phenome-wide scans with 25 CVD related phenotypes (n = 511,700 individuals). We identify 35 lipid-species-associated loci (P &lt;5 ×10−8), 10 of which associate with CVD risk including five new loci-COL5A1, GLTPD2, SPTLC3, MBOAT7 and GALNT16 (false discovery rate&lt;0.05). We identify loci for lipid species that are shown to predict CVD e.g., SPTLC3 for CER(d18:1/24:1). We show that lipoprotein lipase (LPL) may more efficiently hydrolyze medium length triacylglycerides (TAGs) than others. Polyunsaturated lipids have highest heritability and genetic correlations, suggesting considerable genetic regulation at fatty acids levels. We find low genetic correlations between traditional lipids and lipid species. Our results show that lipidomic profiles capture information beyond traditional lipids and identify genetic variants modifying lipid levels and risk of CVD