The Macrophage Activator GcMAF-RF Enhances the Antitumor Effect of Karanahan Technology through Induction of M2–M1 Macrophage Reprogramming

Figure 2

Characterization of the phenotype and synthesis of specific pro- and anti-inflammatory factor mRNAs in the peritoneal macrophages (PMs) isolated from mice, intact and activated with lipopolysaccharide (LPS) and GcMAF-RF. (a) The content of CD11b+Ly-6C+ and CD11b+Ly-6G+ cells (markers characteristic of myeloid suppressor cells) in populations of peritoneal macrophages isolated from intact mice and exposed to medium supplemented with either LPS or GcMAF-RF, relative to that in untreated cells (indicated by a red line). Floating bars (min to max); line at median. The evaluation experiment was performed with two mice in each group. (b) Relative levels of mRNA expression of some pro- and anti-inflammatory factor genes in peritoneal macrophages of intact animals activated with GcMAF-RF and LPS compared with peritoneal macrophages of intact animals not treated with activators (their expression level = 1; the Y-axis is logarithmic). The black asterisk denotes the significant difference from intact cells; the hash indicates the significant difference between cells activated with lipopolysaccharide (LPS) and GcMAF-RF inducers, ; Mann–Whitney U test. Floating bars (min to max); line at median; n = 5, pooled sample. (c) Graphical representation of the relative levels of mRNA expression of some pro- and anti-inflammatory factor genes in the population of peritoneal macrophages, isolated from an intact mouse, after their incubation in medium, medium containing lipopolysaccharide (LPS), or medium containing group-specific macrophage activator (GcMAF-RF). Evaluation of peritoneal macrophage phenotype changes after treatments. The figure schematically shows macrophages. An increase in the number of schematic cells indicates an increase in the number of CD11b+Ly-6C+ monocytic cells among peritoneal macrophages. gray color denotes the M1/M2 phenotype of cells releasing both pro- and anti-inflammatory cytokines; white color denotes the normal M0 phenotype. A factor (cytokine) name without an arrow means its presence in a sample. A crossed-out cytokine name means its absence in a sample. The presence/absence in a sample was assessed by polymerase chain reaction with specific primers. Relative levels of mRNA expression of pro- and anti-inflammatory cytokine genes in LPS/GcMAF-RF-treated peritoneal macrophages compared with those in intact peritoneal macrophages were assessed by Real-time PCR. A factor (cytokine) name with an up (down) arrow means an increase (decrease) in a relative level of cytokine mRNA expression compared with that in intact peritoneal macrophages.

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