Assessing the Impact of BCG Vaccine and Fertile Hydatid Cyst Fluid in Classical and Alternative Macrophage Polarization

Abstract

Classical macrophages (M1) and alternative macrophages (M2) are responsible for various functions in order to maintain homeostasis. The BCG vaccine and hydatid cyst fluid can be examples of stimulants which can cause M1 and 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 is to verify the polarization of macrophages which was induced by BCG vaccine and hydatid cyst fluid by studying the gene expression of aforementioned markers. Mononuclear normal human peripheral blood cells were separated by using ficoll 1077 and gradient concentration. Monocyte populations were separated by adhesion technique from cultured cells in complete tissue culture medium. Monocyte derived macrophages were treated by BCG intravesical vaccine and fertile hydatid cyst fluid for 8 h. The RT-PCR was performed to confirm the polarization. The expression of amplified genes were compared with control groups. Amplification of TNFα and IFNγ genes in M1 and TGFβ and IL4 in M2 group confirmed the polarization procedure. Also, lack of PCR product in negative control samples indicates either 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.