Assessing the gene expression of IL4, TNFα, TGFβ and IFNγ in studying M1 and M2 macrophages derived from human monocyte

Classical macrophages (M1) and alternative macrophages (M2) are responsible for various functions in order to maintain homeostasis. BCG vaccine and hydatid cyst fluid can be examples of stimulants which can cause M1and M2 macrophage polarization. Evaluating the expression of markers such as IFNγ and TNFα for M1 phenotype and TGFβ and IL4 for M2 phenotype is one of the confirmatory ways of polarization. The purpose of this study was to verify the polarization of macrophages which were induced by BCG vaccine and hydatid cyst fluid by studying the gene expression of aforementioned markers. Diluted heparin fresh venous blood was layered by ficol 1077 and then peripheral blood mononuclears were separated by gradient concentration. Monocyte populations were separated by adhesion technique from floating cells in RPMI medium. Derived macrophages were treated by BCG vaccine and hydatid cyst fluid for 8 hours. RT-PCR was performed for confirming the polarization and the expression of amplified genes were compared with control groups. Amplification of TNFα and IFNγ genes in M1 group and TGFβ and IL4 in M2 group confirmed the polarization procedure. Also, lack of PCR product in negative control samples indicates the absence or a very low expression of this gene in control group. The ability of these stimulants in macrophage polarization can be used in experimental studies. It is also possible to take advantage of this model to achieve useful goals in cell therapy and develop more efficient therapeutic methods in inflammatory diseases and cancer immunotherapy.