Microglia in Close Vicinity of Glioma Cells: Correlation Between Phenotype and Metabolic Alterations

Microglia are immune cells within the central nervous system. In brain-developing tumors, gliomas are able to silence the defense and immune functions of microglia, a phenomenon which strongly contributes to tumor progression and treatment resistance. Being activated and highly motile, microglia infiltrate tumors and secrete macrophagic chemoattractant factors. Thereafter, the tumor cells shut down their immune properties and stimulate the microglia to release tumor growth-promoting factors. The result of such modulation is that a kind of symbiosis occurs between microglia and tumor cells, in favor of tumor growth. However, little is known about microglial phenotype and metabolic modifications in a tumoral environment. Co-cultures were performed using CHME5 microglia cells grown on collagen beads or on coverslips and placed on monolayer of C6 cells, limiting cell/cell contacts. Phagocytic behavior and expression of macrophagic and cytoskeleton markers were monitored. Respiratory properties and energetic metabolism were also studied with regard to the activated phenotype of microglia. In co-cultures, transitory modifications of microglial morphology and metabolism were observed linked to a concomitant transitory increase of phagocytic properties. Therefore, after 1 h of co-culture, microglia were activated but when longer in contact with tumor cells, phagocytic properties appear silenced. Like the behavior of the phenotype, microglial respiration showed a transitory readjustment although the mitochondria maintained their perinuclear relocation. Nevertheless, the energetic metabolism of the microglia was altered, suggesting a new energetic steady state. The results clearly indicate that like the depressed immune properties, the macrophagic and metabolic status of the microglia is quickly driven by the glioma environment, despite short initial phagocytic activation. Such findings question the possible contribution of diffusible tumor factors to the microglial metabolism.

[1]  M. Lesniak.  Faculty of 1000 evaluation for Malignant glioma biology: role for TGF-beta in growth, motility, angiogenesis, and immune escape. , 2012 .

[2]  A. Wesołowska,et al.  Microglia-derived TGF-β as an important regulator of glioblastoma invasion—an inhibition of TGF-β-dependent effects by shRNA against human TGF-β type II receptor , 2008, Oncogene.

[3]  P. Magistretti,et al.  Induction of brain aquaporin 9 (AQP9) in catecholaminergic neurons in diabetic rats , 2008, Brain Research.

[4]  K. Moore,et al.  Designer macrophages: oxidative metabolism fuels inflammation repair. , 2006, Cell metabolism.

[5]  D. Greaves,et al.  Oxidative metabolism and PGC-1beta attenuate macrophage-mediated inflammation. , 2006, Cell metabolism.

[6]  S. Kaneko,et al.  Monocyte chemoattractant protein-1 gene delivery enhances antitumor effects of herpes simplex virus thymidine kinase/ganciclovir system in a model of colon cancer , 2006, Cancer Gene Therapy.

[7]  E. González-Rey,et al.  Role of vasoactive intestinal peptide in inflammation and autoimmunity. , 2005, Current opinion in investigational drugs.

[8]  A. Heimberger,et al.  Immunotherapy for human glioma: innovative approaches and recent results , 2005, Expert review of anticancer therapy.

[9]  B. Badie,et al.  Impaired capacity for upregulation of MHC class II in tumor‐associated microglia , 2005, Glia.

[10]  Behnam Badie,et al.  Microglia function in brain tumors , 2005, Journal of neuroscience research.

[11]  M. Rigoulet,et al.  Contribution of the Phosphorylable Complex I in the Growth Phase-Dependent Respiration of C6 Glioma Cells in Vitro , 2003, Journal of bioenergetics and biomembranes.

[12]  R. Jaenisch,et al.  HIF-1α Is Essential for Myeloid Cell-Mediated Inflammation , 2003, Cell.

[13]  B. Scheithauer,et al.  Microglia in brain tumors , 2002, Glia.

[14]  U. Hanisch,et al.  Microglia as a source and target of cytokines , 2002, Glia.

[15]  Walter Heindel,et al.  Phosphorus‐31 MR spectroscopy of normal adult human brain and brain tumours , 2002, NMR in biomedicine.

[16]  F. Aloisi.  Immune function of microglia , 2001, Glia.

[17]  M. Hoehn,et al.  1H‐ and 31P‐MR spectroscopy of primary and recurrent human brain tumors in vitro: malignancy‐characteristic profiles of water soluble and lipophilic spectral components , 2001, NMR in biomedicine.

[18]  Behnam Badie,et al.  Role of microglia in glioma biology , 2001, Microscopy research and technique.

[19]  M. Platten,et al.  Malignant glioma biology: Role for TGF‐β in growth, motility, angiogenesis, and immune escape , 2001 .

[20]  T. Bates,et al.  Activation of murine microglial cell lines by lipopolysaccharide and interferon‐γ causes NO‐mediated decreases in mitochondrial and cellular function , 2001, The European journal of neuroscience.

[21]  Becky,et al.  Dexamethasone-induced abolition of the inflammatory response in an experimental glioma model: a flow cytometry study. , 2000, Journal of neurosurgery.

[22]  U. Gimsa,et al.  Phagocytosis of neuronal or glial debris by microglial cells: Upregulation of MHC class II expression and multinuclear giant cell formation in vitro , 2000, Glia.

[23]  N. Restifo.  Not so Fas: Re-evaluating the mechanisms of immune privilege and tumor escape , 2000, Nature Medicine.

[24]  Daniela Galimberti,et al.  The human astrocytoma cell line U373MG produces monocyte chemotactic protein (MCP)-1 upon stimulation with β-amyloid protein , 2000, Neuroscience Letters.

[25]  M. Ward,et al.  Mitochondrial membrane potential and neuronal glutamate excitotoxicity: mortality and millivolts , 2000, Trends in Neurosciences.

[26]  Philippe Diolez,et al.  Mitochondrial impairment and recovery after heat shock treatment in a human microglial cell line , 2000, Neurochemistry International.

[27]  G. Nagashima,et al.  Fluorescence automatic cell sorter and immunohistochemical investigation of CD68-positive cells in meningioma , 1999, Clinical Neurology and Neurosurgery.

[28]  P. Newsholme,et al.  Metabolic response of macrophages to injury promoted by the activated complement system , 1999, Cell biochemistry and function.

[29]  M. Graeber,et al.  Microglia only weakly present glioma antigen to cytotoxic T cells , 1999, International Journal of Developmental Neuroscience.

[30]  V. Nguyen,et al.  Class II transactivator and class II MHC gene expression in microglia: modulation by the cytokines TGF‐β, IL‐4, IL‐13 and IL‐10 , 1999, European journal of immunology.

[31]  M. Rigoulet,et al.  Energetic and Morphological Plasticity of C6 Glioma Cells Grown on 3-D Support; Effect of Transient Glutamine Deprivation , 1998, Journal of bioenergetics and biomembranes.

[32]  P. Canioni,et al.  Compartmentation of Lactate and Glucose Metabolism in C6 Glioma Cells , 1998, The Journal of Biological Chemistry.

[33]  P. Canioni,et al.  Glucose and Lactate Metabolism in C6 Glioma Cells: Evidence for the Preferential Utilization of Lactate for Cell Oxidative Metabolism , 1998, Developmental Neuroscience.

[34]  H. Rottenberg,et al.  Quantitative assay by flow cytometry of the mitochondrial membrane potential in intact cells. , 1998, Biochimica et biophysica acta.

[35]  M. Bachelet,et al.  Macrophages and heat shock proteins. , 1998, Research in immunology.

[36]  Paul Canioni,et al.  Metabolic and cellular characterization of immortalized human microglial cells under heat stress , 1998, Neurochemistry International.

[37]  R. Myers,et al.  Early metabolic alterations in edematous perihematomal brain regions following experimental intracerebral hemorrhage. , 1998, Journal of neurosurgery.

[38]  R. Sobel,et al.  Effects of Phorbol Myristate Acetate (PMA) on Functions of Macrophages and Microglia In Vitro , 1998, Neurochemical Research.

[39]  D. Peterson,et al.  Mechanism of Cellular 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐Diphenyltetrazolium Bromide (MTT) Reduction , 1997, Journal of neurochemistry.

[40]  P. Fernandes,et al.  Depolymerization of macrophage microfilaments prevents induction and inhibits activity of nitric oxide synthase. , 1996, European journal of cell biology.

[41]  G. Yehia,et al.  Differential expression of heat shock 70 proteins in primary cultures from rat cerebellum , 1996, Brain Research.

[42]  O. Kempski,et al.  Glioblastoma-associated circulating monocytes and the release of epidermal growth factor. , 1996, Journal of neurosurgery.

[43]  H. Hattori,et al.  Increased neurons containing neuronal nitric oxide synthase in the brain of a hypoxic-ischemic neonatal rat model , 1996, Brain and Development.

[44]  D. Aunis,et al.  Chromogranin a triggers a phenotypic transformation and the generation of nitric oxide in brain microglial cells , 1996, Neuroscience.

[45]  A. Cossarizza,et al.  Control of apoptosis by the cellular ATP level , 1996, FEBS letters.

[46]  T. Möller,et al.  Phagocytozing ameboid microglial cells studied in a mouse corpus callosum slice preparation , 1996, Glia.

[47]  J. Saunders,et al.  One-Dimensional Phosphorus-31 Chemical Shift Imaging of Human Brain Tumors , 1995, Investigative radiology.

[48]  P. Magistretti,et al.  Glutamate uptake into astrocytes stimulates aerobic glycolysis: a mechanism coupling neuronal activity to glucose utilization. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[49]  J. Damoiseaux,et al.  Rat macrophage lysosomal membrane antigen recognized by monoclonal antibody ED1. , 1994, Immunology.

[50]  D. Giulian,et al.  Phagocytic microglia release cytokines and cytotoxins that regulate the survival of astrocytes and neurons in culture , 1994, Neurochemistry International.

[51]  F. Wouterlood,et al.  Phagocytic activity of macrophages and microglial cells during the course of acute and chronic relapsing experimental autoimmune encephalomyelitis , 1994, Journal of neuroscience research.

[52]  A. le Pape,et al.  31P and 13C nuclear magnetic resonance studies of macrophages. , 1994, ImmunoMethods.

[53]  W. Hickey,et al.  Characterization of microglia and macrophages in the central nervous system of rats: Definition of the differential expression of molecules using standard and novel monoclonal antibodies in normal CNS and in four models of parenchymal reaction , 1993, Glia.

[54]  S. Walenta,et al.  Proliferation‐associated oxygen consumption and morphology of tumor cells in monolayer and spheroid culture , 1992, Journal of cellular physiology.

[55]  P. Canioni,et al.  Comparative 31P and 1H NMR studies on rat astrocytes and C6 glioma cells in culture. , 1992, Biochimie.

[56]  J. Ruíz-Cabello,et al.  Phospholipid metabolites as indicators of cancer cell function , 1992, NMR in biomedicine.

[57]  L. Montagnier,et al.  Human microglial cells: Characterization in cerebral tissue and in primary culture, and study of their susceptibility to HIV‐1 infection , 1991, Annals of neurology.

[58]  Erwin G. Van Meir,et al.  Human glioblastoma cells release interleukin 6 in vivo and in vitro. , 1990, Cancer research.

[59]  G. Kreutzberg,et al.  Cultured microglial cells have a distinct pattern of membrane channels different from peritoneal macrophages , 1990, Journal of neuroscience research.

[60]  D. Tweardy,et al.  Monokine modulation of human astroglial cell production of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor. I. Effects of IL-1 alpha and IL-beta. , 1990, Journal of immunology.

[61]  Y. Chardonnet,et al.  Evaluation of mitochondrial content and activity with nonyl-acridine orange and rhodamine 123: flow cytometric analysis and comparison with quantitative morphometry , 1990, Cytotechnology.

[62]  M. Ratinaud,et al.  10-N nonyl-acridine orange: a fluorescent probe which stains mitochondria independently of their energetic state. , 1989, Biochemical and biophysical research communications.

[63]  W. Steinbrich,et al.  Combined 1H-MR imaging and localized 31P-spectroscopy of intracranial tumors in 43 patients. , 1988, Journal of computer assisted tomography.

[64]  M. Ratinaud,et al.  In situ flow cytometric analysis of nonyl acridine orange-stained mitochondria from splenocytes. , 1988, Cytometry.

[65]  N. Minshew,et al.  31P Nuclear Magnetic Resonance Studies of Phosphoglyceride Metabolism in Developing and Degenerating Brain: Preliminary Observations , 1987, Journal of neuropathology and experimental neurology.

[66]  J. Rothman,et al.  Uncoating ATPase is a member of the 70 kilodalton family of stress proteins , 1986, Cell.

[67]  M. Delivoria-Papadopoulos,et al.  Unique aspects of human newborn cerebral metabolism evaluated with phosphorus nuclear magnetic resonance spectroscopy , 1984, Annals of neurology.

[68]  K. Gollahon,et al.  Macrophages in experimental and human brain tumors. Part 2: studies of the macrophage content of human brain tumors. , 1979, Journal of neurosurgery.

[69]  K. Gollahon,et al.  Macrophages in experimental and human brain tumors. Part 1: Studies of the macrophage content of experimental rat brain tumors of varying immunogenicity. , 1979, Journal of neurosurgery.

[70]  G. Shambaugh,et al.  Fetal fuels II: contributions of selected carbon fuels to oxidative metabolism in rat conceptus. , 1977, The American journal of physiology.

[71]  A. Wesołowska,et al.  Microglia-derived TGF-beta as an important regulator of glioblastoma invasion--an inhibition of TGF-beta-dependent effects by shRNA against human TGF-beta type II receptor. , 2008, Oncogene.

[72]  W. Paulus,et al.  Ki-M1P as a marker for microglia and brain macrophages in routinely processed human tissues , 2004, Acta Neuropathologica.

[73]  R. Jaenisch,et al.  HIF-1alpha is essential for myeloid cell-mediated inflammation. , 2003, Cell.

[74]  S. Gordon.  Alternative activation of macrophages , 2003, Nature Reviews Immunology.

[75]  V. Tugnoli,et al.  Characterization of lipids from human brain tissues by multinuclear magnetic resonance spectroscopy. , 2001, Biopolymers.

[76]  T. Magnus,et al.  Phagocytosis of apoptotic inflammatory cells by microglia and modulation by different cytokines: Mechanism for removal of apoptotic cells in the inflamed nervous system , 2001, Glia.

[77]  J. Portais,et al.  Glucose and glutamine metabolism in C6 glioma cells studied by carbon 13 NMR. , 1996, Biochimie.

[78]  D Giulian,et al.  The impact of microglia-derived cytokines upon gliosis in the CNS. , 1994, Developmental neuroscience.

[79]  S. Pierce,et al.  A case for chaperones in antigen processing. , 1992, Immunology today.

[80]  P. Canioni,et al.  Phosphorus-31 nuclear magnetic resonance of C6 glioma cells and rat astrocytes. Evidence for a modification of the longitudinal relaxation time of ATP and Pi during glucose starvation. , 1991, European journal of biochemistry.

[81]  K. H. Ng,et al.  HIV receptors within the brain: a study of CD4 and MHC-II on human neurons, astrocytes and microglial cells. , 1991, Research in virology.

[82]  L. Kunz,et al.  Interrelationship among morphology, metabolism, and proliferation of tumor cells in monolayer and spheroid culture. , 1989, Advances in experimental medicine and biology.

[83]  J. Frelinger,et al.  Response of glioma cells to interferon‐gamma: increase in class II RNA, protein and mixed lymphocyte reaction‐stimulating ability , 1985, European journal of immunology.

[84]  C. Hackenbrock,et al.  Continuous measurement of adenosine triphosphate with firefly luciferase luminescence. , 1979, Methods in enzymology.