High-Density Lipoprotein Reduction Differentially Modulates to Classical and Nonclassical Monocyte Subpopulations in Metabolic Syndrome Patients and in LPS-Stimulated Primary Human Monocytes In Vitro

Figures

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  Figure 1. Percentage of classical, intermediate, and nonclassical monocytes in metabolic syndrome patients and control subjects. Representative flow cytometry dot plots showing the percentage of classical (CM), intermediate (IM), and nonclassical monocytes (NCM) in control subjects (a) and patients with metabolic syndrome (b). The CM percentage is significantly decreased in metabolic syndrome patients as compared to controls (c). The IM percentage showed no significant differences between metabolic syndrome patients and controls (d). The NCM percentage is significantly increased in metabolic syndrome patients as compared to controls (e). For gating strategy, white blood cells were firstly gated for singlets on a FSC-H/FSC-A density plot. Then, lymphocyte, granulocyte, and monocyte populations were gated on a FSC-A/SSC-A plot. On the monocyte gate, living cells were further gated using the Live/Dead Aqua stain. Living monocytes were then gated to determine CD14- and CD16-positive expression and identify monocyte subpopulations as follows: CD14highCD16−, classical monocytes; CD14highCD16+, intermediate monocytes; and CD14lowCD16+, nonclassical monocytes. In panels (c)–(e), data are expressed…

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  Figure 2. Effect of HDL on LPS-stimulated primary human monocytes in vitro. Representative polyacrylamide gel showing a metabolic syndrome patient's serum sample in which HDL was totally removed (−HDL) and then reconstituted with 0.77 mmol/L (30 mg/dL) or 1.55 mmol/L (60 mg/dL) HDL (a). As compared to untreated cells, LPS stimulation induced reduction in the CM percentage in low HDL levels (zero and 30 mg/dL) (b, left and middle panels, resp.). In contrast, the effect of LPS on the CM percentage was abolished in 60 mg/dL HDL that resembled a high HDL concentration (b, right panel). LPS stimulation did not significantly modify the IM percentage neither in low nor in high HDL concentrations (c, left, middle and right panels, resp.). As compared to untreated cells, LPS stimulation increased the NCM percentage in zero and 30 mg/dL HDL (d, left and middle panels, resp.). On the contrary, the effect of LPS on the NCM percentage was abolished in high HDL concentrations (d, right panel). As compared to untreated cells, LPS stimulation increased IL-1β production in primary human monocytes cultured in low HDL concentrations (e, left and middle panels, resp.). In contrast, the effect of LPS on IL-1β…

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  Figure 3. Serum levels of IL-1β in the study patients according to different metabolic syndrome risk factors. (a) Metabolic syndrome patients displaying central obesity (n = 39) showed similar IL-1β serum levels than did metabolic syndrome patients that had a normal waist circumference (n = 5). (b) The serum levels of IL-1β did not show significant differences in metabolic syndrome patients with elevated blood pressure (n = 6) as compared to metabolic syndrome patients with normal blood pressure (n = 38). (c) Metabolic syndrome patients showing fasting hyperglycemia (n = 8) exhibited similar IL-1β circulating levels than did metabolic syndrome patients with normal glycemic values (n = 36). (d) The serum levels of IL-1β tended to increase in metabolic syndrome patients with hypertriglyceridemia (n = 23) but did not show significant differences with respect to metabolic syndrome patients showing normal triglyceride values (n = 21). (e) In contrast, IL-1β was significantly increased in metabolic syndrome patients with HDL low levels (n = 34) as compared to metabolic syndrome patients exhibiting normal HDL values (n = 10). Data are expressed as mean ± standard deviation. Significant…