ABSTRACT
BACKGROUND: Macrophages are a functionally heterogeneous cell population and depending on microenvironments they polarize in two main groups: M1 and M2. Glutamic acid and glutamate receptors may participate in the regulation of macrophage plasticity. To investigate the role of glutamatergic systems in macrophages physiology, we performed the transfection of mGluR5 cDNAs into RAW-264.7 cells. RESULTS: Comparative analysis of modified (RAW-mGluR5 macrophages) and non-modified macrophages (RAW-macrophages) has shown that the RAW-mGluR5 macrophages absorbed more glutamate than control cells and the amount of intracellular glutamate correlated with the expression of excitatory amino acid transporters -2 (EAAT-2). Besides, our results have shown that RAW-mGluR5 macrophages expressed a higher level of peroxisome proliferator-activated receptor γ (PPAR-γ) and secreted more IL-10, high mobility group box 1 proteins (HMGB1) and Galectin-3 than control RAW-macrophages. CONCLUSIONS: We propose that elevation of intracellular glutamate and expression of mGluR5 may initiate the metabolic rearrangement in macrophages that could contribute to the formation of an immunosuppressive phenotype.
Subject(s)
Cell Plasticity/physiology , Macrophages/physiology , Receptor, Metabotropic Glutamate 5/physiology , Animals , Blotting, Western , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Galectin 3/analysis , Galectin 3/metabolism , Glutamic Acid/analysis , Glutamic Acid/metabolism , HMGB1 Protein/analysis , HMGB1 Protein/metabolism , Interleukin-10/analysis , Interleukin-10/metabolism , Lipopolysaccharides , Mice , Nitric Oxide/metabolism , PPAR alpha/analysis , PPAR alpha/metabolism , Phenotype , RAW 264.7 Cells , Transfection/methodsABSTRACT
Abstract Background Macrophages are a functionally heterogeneous cell population and depending on microenvironments they polarize in two main groups: M1 and M2. Glutamic acid and glutamate receptors may participate in the regulation of macrophage plasticity. To investigate the role of glutamatergic systems in macrophages physiology, we performed the transfection of mGluR5 cDNAs into RAW-264.7 cells. Results Comparative analysis of modified (RAW-mGluR5 macrophages) and non-modified macrophages (RAW-macrophages) has shown that the RAW-mGluR5 macrophages absorbed more glutamate than control cells and the amount of intracellular glutamate correlated with the expression of excitatory amino acid transporters -2 (EAAT-2). Besides, our results have shown that RAW-mGluR5 macrophages expressed a higher level of peroxisome proliferator-activated receptor γ (PPAR-γ) and secreted more IL-10, high mobility group box 1 proteins (HMGB1) and Galectin-3 than control RAW-macrophages. Conclusions We propose that elevation of intracellular glutamate and expression of mGluR5 may initiate the metabolic rearrangement in macrophages that could contribute to the formation of an immunosuppressive phenotype.
Subject(s)
Animals , Mice , Receptor, Metabotropic Glutamate 5/physiology , Cell Plasticity/physiology , Macrophages/physiology , Phenotype , Enzyme-Linked Immunosorbent Assay , Transfection/methods , Cells, Cultured , Lipopolysaccharides , Blotting, Western , Interleukin-10/analysis , Interleukin-10/metabolism , Glutamic Acid/analysis , Glutamic Acid/metabolism , HMGB1 Protein/analysis , HMGB1 Protein/metabolism , Galectin 3/analysis , Galectin 3/metabolism , PPAR alpha/analysis , PPAR alpha/metabolism , RAW 264.7 Cells , Nitric Oxide/metabolismABSTRACT
Human T lymphocytes express both ionotropic and metabotropic glutamate receptors that control immune responses, cell activation, maturation and death. In this study, we examined the effect of N-methyl-D-aspartate (NMDA) and sigma1-receptor ligands on the secretion of the proinflammatory chemokine interleukin 8 (IL-8) and the anti-inflammatory cytokine interleukin 10 (IL-10) in human leukemia Jurkat cells and peripheral blood lymphocytes (PBLs). We have shown that NMDA increased IL-8 and decreased IL-10 secretion and that sigma-ligands modulated the action of NMDA. Moreover, the effects of NMDA and sigma-ligands were interrelated with the nitric oxide (NO) content, suggesting that the intracellular concentration of NO could play a major role in the synthesis of cytokines. Western blots against the NR2A and NR2B subunits of the NMDA glutamate receptor revealed that long-term (48 h) treatment of PBLs with glutamate at concentrations within normal plasma levels (1 x 10(-5)M), in contrast to low concentrations (0.3 x 10(-6)M), downregulates the NR2A subunit, probably by internalization. Furthermore, we found that PBLs with noninternalized NR2A secreted less IL-10 than lymphocytes with downregulated NR2A; under these conditions, the transcriptional activity of NF-kappaB was increased whereas the transcriptional activity of c-Fos was decreased. These findings implicate that the activities of NF-kappaB and c-Fos control the expression of the IL8 and IL10 genes, depending on the subunit composition of the NMDA receptor. In conclusion, we suggest that lymphocytes express an active NMDA receptor only in a low-glutamate milieu.
