Alf Åge Pettersen

Circulating levels of IL-18 are significantly influenced by the IL-18 +183 A/G polymorphism in coronary artery disease patients with diabetes type 2 and the metabolic syndrome: an Observational Study

BackgroundIncreased IL-18 serum levels have been associated with diabetes type 2, metabolic syndrome and the severity of atherosclerosis. The present study investigated the presence and influence of IL-18 genetic variants on gene- and protein expression in stable coronary artery disease (CAD) patients.MethodsThe +183 A/G (rs 5744292), -137 G/C (rs 187238) and -607 C/A (rs 1946518) polymorphisms were determined in 1001 patients with angiographically verified stable CAD, and in 204 healthy controls. IL-18 gene-expression was measured in circulating leukocytes in 240 randomly selected patients. Circulating IL-18 and IL-18 binding protein levels were measured immunologically in all patients.ResultsThe +183 G-allele associated significantly with lower serum levels of IL-18 (p = 0.002, adjusted for age, glucose, body mass index and gender) and a 1.13- fold higher IL-18 gene-expression (p = 0.010). No influence was observed for the -137 G/C and -607 C/A polymorphisms. The IL-18 binding protein levels were not influenced by IL-18 genotypes. IL-18 levels were significantly higher in men as compared to women, and in patients with diabetes type 2 and metabolic syndrome compared to those without (p ≤ 0.001, all). The reduction in IL-18 levels according to the +183 G-allele was 3-4 fold more pronounced in diabetes and metabolic syndrome as compared to unaffected patients.Finally, the +183 AA genotype was more frequent in patients with hypertension (p = 0.042, adjusted for age, body mass index and gender).ConclusionThe reduction in serum IL-18 levels across increasing numbers of +183G-alleles was especially apparent in patient with diabetes type 2 and metabolic syndrome, suggesting a beneficial GG genotype in relation to cardiovascular outcome in these patients.Clinical Trial Registration NumberClinicalTrials.gov: NCT00222261

The Co-Existence of the IL-18+183 A/G and MMP-9 -1562 C/T Polymorphisms Is Associated with Clinical Events in Coronary Artery Disease Patients

Objective Interleukin (IL)-18 has been associated with severity of atherosclerosis and discussed to predict cardiovascular (CV) events. We have previously shown that the IL-18+183 G-allele significantly reduces IL-18 levels. This study was aimed to investigate the prognostic significance of the IL-18+183 A/G polymorphism (rs5744292), single and in coexistence with the matrix metalloproteinase (MMP)-9 -1562 C/T (rs3918242) polymorphism, in patients with stable coronary artery disease (CAD). Serum levels of IL-18, MMP-9 and tissue inhibitor of matrix metalloproteinase (TIMP)-1 were additionally assessed. Methods 1001 patients with angiographically verified CAD were genotyped and the biomarkers were measured accordingly. After two years follow-up, 10.6% experienced new clinical events; acute myocardial infarction (AMI), stroke, unstable angina pectoris and death. Results The IL-18+183 G-allele associated with 35% risk reduction in composite endpoints after adjusting for potential covariates (p = 0.044). The IL-18+183 AA/MMP-9 -1562 CT/TT combined genotypes associated with a significant increase in risk of composite endpoints (OR = 1.87; 95% CI = 1.13–3.11, p = 0.015, adjusted). Patients with clinical events presented with significantly higher IL-18 levels as compared to patients without (p = 0.011, adjusted). The upper tertile of IL-18 levels associated with an increase in risk of AMI as compared to lower tertiles (OR = 2.36; 95% CI = 1.20–4.64, p = 0.013, adjusted). Conclusion The IL-18+183 A/G polymorphism, single and in combination with MMP-9 genotypes, may influence the risk of clinical events in stable CAD patients.

The MMP-9 -1562 C/T Polymorphism in the Presence of Metabolic Syndrome Increases the Risk of Clinical Events in Patients with Coronary Artery Disease

Background and Objectives Elevated levels of matrix metalloproteinase (MMP)-9 have been associated with the metabolic syndrome (MetS) and cardiovascular events. The MMP-9 −1562 C/T polymorphism has furthermore been shown as a risk factor for coronary artery disease (CAD). The non-favourable cardiometabolic state in MetS may increase the risk. We aimed to investigate the influence of MMP-9 −1562 C/T polymorphism in subjects with CAD and MetS. Methods Patients (n = 1000) with verified CAD stratified in Mets +/− (n = 244/756), were analyzed for the MMP-9 −1562 C/T polymorphism and related to clinical events after 2 years follow-up. Serum levels of total MMP-9 and tissue inhibitor of matrix metalloproteinases (TIMP)-1were analyzed in all, whereas MMP-9 activity, extracellular matrix metalloproteinase inducer (EMMPRIN), and expression of the two genes were analyzed in a subset of 240 randomly selected patients. Results Totally, 106 clinical endpoints were recorded. In MetS; the T-allele associated with 5.5 fold increase in event rate (p<0.0001), increased with number of MetS components, a 117% increase in total MMP-9 levels (TT homozygous, p = 0.05), significantly higher total- and endogenous active MMP-9 and TIMP-1 levels (p<0.01 all), and EMMPRIN was inversely correlated with pro- and endogenous active MMP-9 (p<0.05, both). In non-MetS; the T-allele was not associated with new events, nor higher MMP-9 levels. EMMPRIN was significantly correlated with total MMP-9 and TIMP-1 (p<0.01, both) and the two genes were inter-correlated (p<0.001). Conclusion In CAD patients with MetS, the MMP-9 T-allele increased the risk of clinical events, probably mediated through elevated MMP-9 levels and altered MMP-9 regulation.

Fractalkine and its receptor (CX3CR1) in patients with stable coronary artery disease and diabetes mellitus.

BACKGROUND Fractalkine and its receptor CX3CR1 are associated with atherosclerosis. In vitro studies have shown increased expression of fractalkine in endothelial and vascular smooth muscle cells when stimulated with a high concentration of glucose. Increased serum levels of fractalkine have been shown in patients with type 2 diabetes mellitus (T2DM) and also in unstable coronary artery disease (CAD) patients. We investigated whether CAD patients with T2DM or metabolic syndrome have increased circulating and gene expression levels of fractalkine compared to CAD patients without these conditions. METHODS Serum levels of fractalkine were analyzed by the enzyme-linked immunosorbent assay (ELISA) method in 1001 patients with angiographically verified CAD, of which 200 had T2DM and 244 had metabolic syndrome. All patients were taking aspirin as an antithrombotic treatment. Gene expression of fractalkine and CX3CR1 in circulating leukocytes was explored in a subset of patients (n=168). RESULTS We found no significant difference in circulating levels of fractalkine in patients with T2DM [653 (556, 775) pg/mL] compared to patients without T2DM [646 (553, 761) pg/mL], p=0.50. There was also no difference between patients with and without metabolic syndrome (p=0.60). Fractalkine was not expressed in circulating leukocytes, and CX3CR1 was not expressed differently between any of the groups (p=0.13 and p=0.32, respectively). Smokers had lower fractalkine levels (p<0.001), and patients on angiotensin II receptor blockers had higher levels (p=0.047) compared to nonaffected patients. CONCLUSIONS In the present CAD population, no differences in circulating levels of fractalkine or expression levels of CX3CR1 were observed between patients with and without T2DM, or with and without metabolic syndrome, which may be related to their underlying disease.

Combined Elevated Levels of the Proinflammatory Cytokines IL-18 and IL-12 Are Associated with Clinical Events in Patients with Coronary Artery Disease: An Observational Study

BACKGROUND Interleukin (IL)-18 in synergy with IL-12 is critical in the initiation and progression of Th-1-type responses. IL-18 and IL12 elevation has been associated with atherosclerosis, and their interaction is hypothesized to partly be driven by glucose. We aimed to explore if simultaneous elevation of IL-18 and IL-12, as related to glucose levels, would influence the prognosis in coronary artery disease (CAD). METHOD Patients (n = 1001) with angiographically verified stable CAD were investigated (78% men, mean age 62 years, 20% current smokers). IL-18 and IL-12 were measured by conventional ELISA methods. High fasting glucose (FG) was defined as the 75 percentile, that is, >6.2 mmol/L. RESULTS After 2-year follow-up, 100 cardiovascular endpoints (fatal and nonfatal acute myocardial infarction, unstable angina, and stroke) were recorded. Subjects with simultaneous (not separate) levels in upper tertiles of both markers were at higher risk of cardiovascular events, compared to subjects in lowest tertile of both (odds ratio = 1.70, 95% confidence interval 1.11-2.61, adjusted P = 0.016), with no influence of high FG. Hyperglycemia associated with higher IL-18 levels (adjusted P = 0.009) and IL-12 levels was considerably lower in current smokers (adjusted P < 0.001). CONCLUSION Simultaneous elevated circulating levels of IL-18 and IL-12 increased the event rate after 2 years in CAD patients, independent of hyperglycemia.

Markers of neutrophil extracellular traps are associated with adverse clinical outcome in stable coronary artery disease

Background Neutrophil extracellular traps, comprising chromatin and granule proteins, have been implicated in atherothrombosis. Design and methods We investigated whether the circulating neutrophil extracellular traps markers, double-stranded DNA and myeloperoxidase-DNA were associated with clinical outcome and hypercoagulability in patients with stable coronary artery disease. Patients with angiographically verified stable coronary artery disease (n = 1001) were included. Follow-up was 2 years, recording 106 clinical endpoints (unstable angina, non-haemorrhagic stroke, myocardial infarction or death). Serum collected at baseline was used to determine double-stranded DNA and myeloperoxidase-DNA levels. Results The neutrophil extracellular traps markers were weakly intercorrelated (r = 0.103, P = 0.001). Patients with the highest quartile of double-stranded DNA had weakly but significantly elevated hypercoagulability markers (prothrombin fragment 1+2, D-dimer, free and total tissue factor pathway inhibitor (P < 0.001 for all)). Men, smokers, patients with metabolic syndrome and patients with a previous myocardial infarction had significantly elevated double-stranded DNA levels (P ≤ 0.002 for all). Significantly higher double-stranded DNA levels were observed in the group experiencing a clinical endpoint compared to the group without (P = 0.019). When categorising double-stranded DNA into quartiles, a distinct cut-off between the lowest and upper three quartiles was observed. Adjusting for relevant covariates, patients in the upper three quartiles had an odds ratio of 2.01 (95% confidence interval 1.12, 3.58, P = 0.019) for experiencing a clinical endpoint. Myeloperoxidase-DNA was not significantly associated with clinical outcome or hypercoagulability. Conclusions Double-stranded DNA levels were significantly related to adverse clinical outcome after 2 years, but only weakly associated with hypercoagulability. These observations suggest that the detrimental effects of neutrophil extracellular traps in coronary artery disease might extend beyond those related to hypercoagulability.