Molecular Diversity of Macrophages in Allergic Reaction: Comparison between the Allergenic Modes; Th1- and -Th2-Derived Immune Conditions
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 Alu...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Tehran University of Medical Sciences
2015-10-01
|
| Series: | Iranian Journal of Allergy, Asthma and Immunology |
| Subjects: | |
| Online Access: | https://ijaai.tums.ac.ir/index.php/ijaai/article/view/566 |
| _version_ | 1811258382145814528 |
|---|---|
| author | Mozhdeh Bagheri Yupeng Dong Masao Ono |
| author_facet | Mozhdeh Bagheri Yupeng Dong Masao Ono |
| author_sort | Mozhdeh Bagheri |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T18:13:28Z |
| format | Article |
| id | doaj.art-deb3ac80603442f1bf7958c2ff0913a6 |
| institution | Directory Open Access Journal |
| issn | 1735-1502 1735-5249 |
| language | English |
| last_indexed | 2024-04-12T18:13:28Z |
| publishDate | 2015-10-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Allergy, Asthma and Immunology |
| spelling | doaj.art-deb3ac80603442f1bf7958c2ff0913a62022-12-22T03:21:45ZengTehran University of Medical SciencesIranian Journal of Allergy, Asthma and Immunology1735-15021735-52492015-10-01143530Molecular Diversity of Macrophages in Allergic Reaction: Comparison between the Allergenic Modes; Th1- and -Th2-Derived Immune ConditionsMozhdeh Bagheri0Yupeng Dong1Masao Ono2Department 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, JapanDepartment 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, JapanDepartment of Pathology, Tohoku University Graduate School of Medicine, Sendai, JapanActivated 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/566Cell MarkersComplete Freund AdjuvantCytokinesHypersensitivityInflammatoryMacrophages |
| spellingShingle | Mozhdeh Bagheri Yupeng Dong Masao Ono Molecular Diversity of Macrophages in Allergic Reaction: Comparison between the Allergenic Modes; Th1- and -Th2-Derived Immune Conditions Iranian Journal of Allergy, Asthma and Immunology Cell Markers Complete Freund Adjuvant Cytokines Hypersensitivity Inflammatory Macrophages |
| title | Molecular Diversity of Macrophages in Allergic Reaction: Comparison between the Allergenic Modes; Th1- and -Th2-Derived Immune Conditions |
| title_full | Molecular Diversity of Macrophages in Allergic Reaction: Comparison between the Allergenic Modes; Th1- and -Th2-Derived Immune Conditions |
| title_fullStr | Molecular Diversity of Macrophages in Allergic Reaction: Comparison between the Allergenic Modes; Th1- and -Th2-Derived Immune Conditions |
| title_full_unstemmed | Molecular Diversity of Macrophages in Allergic Reaction: Comparison between the Allergenic Modes; Th1- and -Th2-Derived Immune Conditions |
| title_short | Molecular Diversity of Macrophages in Allergic Reaction: Comparison between the Allergenic Modes; Th1- and -Th2-Derived Immune Conditions |
| title_sort | molecular diversity of macrophages in allergic reaction comparison between the allergenic modes th1 and th2 derived immune conditions |
| topic | Cell Markers Complete Freund Adjuvant Cytokines Hypersensitivity Inflammatory Macrophages |
| url | https://ijaai.tums.ac.ir/index.php/ijaai/article/view/566 |
| work_keys_str_mv | AT mozhdehbagheri moleculardiversityofmacrophagesinallergicreactioncomparisonbetweentheallergenicmodesth1andth2derivedimmuneconditions AT yupengdong moleculardiversityofmacrophagesinallergicreactioncomparisonbetweentheallergenicmodesth1andth2derivedimmuneconditions AT masaoono moleculardiversityofmacrophagesinallergicreactioncomparisonbetweentheallergenicmodesth1andth2derivedimmuneconditions |