Tehran University of Medical Sciences Iranian Journal of Allergy, Asthma and Immunology 1735-1502 14 3 2015 10 18 261 272 EN Mozhdeh Bagheri Department of Pathology, Tohoku University Graduate School of Medicine, Sendai, Japan AND Department of Stem Cell Biology and Histology, Tohoku University Graduate School of Medicine, Sendai, Japan Yupeng Dong Department of Pathology, Tohoku University Graduate School of Medicine, Sendai, Japan AND Department of Disability Sciences, Advanced Interdisciplinary Biomedical Functional Tohoku University Graduate School of Medicine, Sendai, Japan Masao Ono Department of Pathology, Tohoku University Graduate School of Medicine, Sendai, Japan 2015 10 18 2015 10 18 Activated macrophages have been classified into classical (M1) and alternative (M2) macrophages. We aimed to establish a method to yield enough number of macrophages to analyze their molecular, biological and immunological functions. We used drugs; adjuvant albumin from chicken egg whites—Imject Alum (OVA- Alum) and OVA Complete Freund Adjuvant (OVA-CFA), to induce macrophages to M2 and M1 respectively.We analyzed the phenotype of purified macrophages induced under these immune conditions, using flow cytometry (FACS) to detect cell-surface molecules and the enzyme-linked immunosorbent assay (ELISA) was used to detect cytokines. The cDNA microarray was employed to measure changes in expression level of cell surface protein between M1 and M2 macrophages.Phenotype   analysis   of   purified   macrophages,   induced   under   these   immune   conditions,   showed macrophages induced by OVA-Alum was almost M2 while the proportion of M1 macrophages induced by OVA-CFA was significantly higher. The results also showed higher expression level of macrophage galactose N- acetyl-galactosamine specific lectin-2 protein (MGL1/2-PE), a known M2 macrophage marker, on the surface of Alum-induced  macrophages.  On  the  basis  of  these  preliminary  data,  ELISA  results  revealed  that  after macrophage stimulation with lipopolysaccharides (LPS), the level of interleukin (IL)-10 produced by Alum- induced macrophages was higher than the level of IL-10 produced by CFA-induced macrophages. In contrast, the level of tumor necrosis factor-alpha (TNF-α) produced by CFA-induced macrophages was higher than Alum-induced macrophages.The cDNA microarray confirmed previous results and suggest immunoglobulin-like type 2 receptor alpha (Pilra) as a new marker for M1, macrophage galactose N-acetylgalactosamine-specific lectin 2 (Mgl2) as M2 macrophages marker. https://ijaai.tums.ac.ir/index.php/ijaai/article/view/566 https://ijaai.tums.ac.ir/index.php/ijaai/article/download/566/451