CMV Infection Is Directly Related to the Inflammatory Status in Chronic Heart Failure Patients

Figures

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  Figure 1. Cytokine levels in the two studied groups. Cytokine concentrations were measured using Luminex multiplex technology. Levels of cytokines in CHF (n = 40) and controls (n = 40) are illustrated in the box plots. Differences between groups in the levels of cytokines were compared using the Mann-Whitney Test, p-values are depicted in the panels. Outlier values are represented by circles and extreme values by stars, calculated by adding 1.5 and 3 times the IR to the 75th percentile, respectively.

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  Figure 2. Levels of cytokines analyzed in CHF with respect to the different degrees of HF classified according NYHA (New York Heart Association) in CMV- and CMV+ patients. Outlier values are represented by circles and extreme values by stars, calculated by adding 1.5 and 3 times the IR to the 75th percentile, respectively. The ANOVA test was used to examine differences between the groups; p-values are depicted in the boxed text; the interaction is between NYHA and CMV serostatus. If significant interactions were observed, comparisons with a Bonferroni correlated post hoc test were performed and p-values are represented in the panels.

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  Figure 3. Relationship between anti-CMV antibody titer with cytokine levels in CHF patients. The correlation of anti-CMV antibody titers and cytokine levels in CHF patients is represented in the dot plots. Spearman’s test was applied to calculate the correlations; the p-value and coefficient of correlation are listed on the panels.

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  Figure 4. Levels of cytokines correlated to CD28null T-lymphocyte populations in CHF patients. Levels of cytokines (pg/mL) in CD4+ (A) and CD8+ T-lymphocytes (B). Spearman’s test was applied to calculate the correlations; the p-value and coefficient of correlation are listed on the panels.

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  Figure 5. Cytokine levels produced in response to different culture conditions in CHF patients according to their CMV-serostatus. PBMCs from CHF patients (CMV-, n = 7; CMV+, n = 9) were cultured for 5 days in medium alone, in medium containing anti-CD3 (1 µg/ml), or in medium containing LPS (1 µg/ml). In the supernatant of the different cultures, the concentrations of the different cytokines were measured using Luminex multiplex technology and analyzed according their CMV-serostatus. Bar graphs summarize the concentrations of the cytokines in the supernatants of the cultures with the different conditions studied. (means ± SEM) from the studied subjects. Paired t-test was used to compare paired means, and p-values are depicted in the panels. *indicates a significant difference (p <0.05) compared to the other groups.

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  Figure 6. Changes in the gene expression profile in CHF patients divided according to their seropositivity to CMV measured by TaqMan™ Array Human Immune Response. Heat map showing differential expression in patients with CHF CMV+ compared to CMV- patients. Green colors indicate under-expression in CMV+ patients and red tones overexpression in these patients. The blue color is indicating the genes used as endogenous control genes in 5 CHF CMV- and 5 CHF CMV+ patients. We used the comparative ddCT method for calculating relative quantitation of gene expression after outlier removal and data normalization based on the endogenous control genes expression using DataAssist software (Thermo Fisher Scientific).