Use of Primary Macrophages for Searching Novel Immunocorrectors

Abstract

In this mini-review, the role of macrophage phenotypes in atherogenesis is considered. Recent studies on distribution of M1 and M2 macrophages in different types of atherosclerotic lesions indicate that macrophages exhibit a high degree of plasticity of phenotype in response to various conditions in microenvironment. The effect of the accumulation of cholesterol, a key event in atherogenesis, on the macrophage phenotype is also discussed. The article presents the results of transcriptome analysis of cholesterol-loaded macrophages revealing genes involved in immune response whose expression rate has changed the most. It turned out that the interaction of macrophages with modified LDL leads to higher expression levels of pro-inflammatory marker TNF-α and antiinflammatory marker CCL18. Phenotypic profile of macrophage activation could be a good target for testing of novel anti-atherogenic immunocorrectors. A number of anti-atherogenic drugs were tested as potential immunocorrectors using primary macrophage-based model.

Keywords: Atherosclerosis; activation; immunocorrectors; macrophage-based test; macrophages; modified LDL; phenotype; transcriptome.

Fig. (1).

Effect of LDL on cytokine gene expression. Monocytes were isolated from whole blood of healthy donors by density gradient followed by selection of CD14+ cells by magnetic separation. Cells were cultured for 7 days. Native or atherogenic LDL was added at a concentration of 100 µg / ml and the cells were incubated for 24 hours. RNA was isolated and gene expression was measured by RT-PCR technique. The figures show the relative expression of the genes TNF-α (n=14) and CCL18 (n=7). As 1, the control gene expression (without LDL) was taken. * Significant differences from native LDL (T-Test).