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

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: CD14^highCD16⁻, classical monocytes; CD14^highCD16⁺, intermediate monocytes; and CD14^lowCD16⁺, nonclassical monocytes. In panels (c)–(e), data are expressed as . Significant differences were estimated by means of performing Student’s t-test. Differences were considered significant when . Diagnosis of metabolic syndrome was performed according to the ATP III criteria, when three of five of the following factors were present: central obesity denoted by a waist circumference greater than 80 cm in women and 90 cm in men, hypertriglyceridemia (circulating triglyceride ), decreased serum values of HDL-cholesterol (serum in men and 50 mg/dL in women), blood pressure higher than 120/80 mmHg, and hyperglycemia (fasting blood ).

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