TGFβ signalling plays an important role in IL4-induced alternative activation of microglia

Journal of Neuroinflammation

Kerstin Krieglstein¹  Björn Spittau²  Xiaolai Zhou³ 

[1] Freiburg Institute for Advanced Studies (FRIAS), Albert-Ludwigs-University Freiburg, Albertstraße 19, Freiburg 79104, Germany;Institute for Anatomy and Cell Biology, Department of Molecular Embryology, Albert-Ludwigs-University Freiburg, Albertstraße 17, Freiburg 79104, Germany;Faculty of Biology, Albert-Ludwigs-University Freiburg, Hauptstraße 1, 79104 Freiburg, Germany

关键词: Ym1;    Arg1;    Alternative activation;    IL4;    TGFβ;    Microglia;   

Others  :  1160268
DOI  :  10.1186/1742-2094-9-210

 received in 2012-02-23, accepted in 2012-08-17,  发布年份 2012

【 摘 要 】

Background

Microglia are the resident immune cells of the central nervous system and are accepted to be involved in a variety of neurodegenerative diseases. Several studies have demonstrated that microglia, like peripheral macrophages, exhibit two entirely different functional activation states, referred to as classical (M1) and alternative (M2) activation. TGFβ is one of the most important anti-inflammatory cytokines and its effect on inhibiting microglia or macrophage classical activation has been extensively studied. However, the role of TGFβ during alternative activation of microglia has not been described yet.

Methods

To investigate the role of TGFβ in IL4-induced microglia alternative activation, both, BV2 as well as primary microglia from new born C57BL/6 mice were used. Quantitative RT-PCR and western blots were performed to detect mRNA and protein levels of the alternative activation markers Arginase1 (Arg1) and Chitinase 3-like 3 (Ym1) after treatment with IL4, TGFβ or both. Endogenous TGFβ release after IL4 treatment was evaluated using the mink lung epithelial cell (MLEC) assay and a direct TGFβ2 ELISA. TGFβ receptor type I inhibitor and MAPK inhibitor were applied to address the involvement of TGFβ signalling and MAPK signalling in IL4-induced alternative activation of microglia.

Results

TGFβ enhances IL4-induced microglia alternative activation by strongly increasing the expression of Arg1 and Ym1. This synergistic effect on Arg1 induction is almost completely blocked by the application of the MAPK inhibitor, PD98059. Further, treatment of primary microglia with IL4 increased the expression and secretion of TGFβ2, suggesting an involvement of endogenous TGFβ in IL4-mediated microglia activation process. Moreover, IL4-mediated induction of Arg1 and Ym1 is impaired after blocking the TGFβ receptor I indicating that IL4-induced microglia alternative activation is dependent on active TGFβ signalling. Interestingly, treatment of primary microglia with TGFβ alone results in up regulation of the IL4 receptor alpha, indicating that TGFβ increases the sensitivity of microglia for IL4 signals.

Conclusions

Taken together, our data reveal a new role for TGFβ during IL4-induced alternative activation of microglia and consolidate the essential functions of TGFβ as an anti-inflammatory molecule and immunoregulatory factor for microglia.

【 授权许可】

   
2012 Zhou et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Vaughan DW, Peters A: Neuroglial cells in the cerebral cortex of rats from young adulthood to old age: an electron microscope study. J Neurocytol 1974, 3:405-429.
• [2]Colton CA, Mott RT, Sharpe H, Xu Q, Van Nostrand WE, Vitek MP: Expression profiles for macrophage alternative activation genes in AD and in mouse models of AD. J Neuroinflammation 2006, 3:27. BioMed Central Full Text
• [3]Colton CA, Wilcock DM: Assessing activation states in microglia. CNS Neurol Disord Drug Targets 2010, 9:174-191.
• [4]Shimizu E, Kawahara K, Kajizono M, Sawada M, Nakayama H: IL-4-induced selective clearance of oligomeric beta-amyloid peptide(1–42) by rat primary type 2 microglia. J Immunol 2008, 181:6503-6513.
• [5]Ponomarev ED, Maresz K, Tan Y, Dittel BN: CNS-derived interleukin-4 is essential for the regulation of autoimmune inflammation and induces a state of alternative activation in microglial cells. J Neurosci 2007, 27:10714-10721.
• [6]Ponomarev ED, Veremeyko T, Barteneva N, Krichevsky AM, Weiner HL: MicroRNA-124 promotes microglia quiescence and suppresses EAE by deactivating macrophages via the C/EBP-alpha-PU.1 pathway. Nat Med 2011, 17:64-70.
• [7]Rojo AI, Innamorato NG, Martin-Moreno AM, De Ceballos ML, Yamamoto M, Cuadrado A: Nrf2 regulates microglial dynamics and neuroinflammation in experimental Parkinson’s disease. Glia 2010, 58:588-598.
• [8]Smith PF: Inflammation in Parkinson’s disease: an update. Curr Opin Investig Drugs 2008, 9:478-484.
• [9]Rubartelli A, Lotze MT: Inside, outside, upside down: damage-associated molecular-pattern molecules (DAMPs) and redox. Trends Immunol 2007, 28:429-436.
• [10]Mosser DM: The many faces of macrophage activation. J Leukoc Biol 2003, 73:209-212.
• [11]Gordon S: Alternative activation of macrophages. Nat Rev Immunol 2003, 3:23-35.
• [12]Gao HM, Hong JS: Why neurodegenerative diseases are progressive: uncontrolled inflammation drives disease progression. Trends Immunol 2008, 29:357-365.
• [13]Giulian D: Ameboid microglia as effectors of inflammation in the central nervous system. J Neurosci Res 1987, 18:155-171. 132–153
• [14]Block ML, Zecca L, Hong JS: Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci 2007, 8:57-69.
• [15]Town T, Nikolic V, Tan J: The microglial “activation” continuum: from innate to adaptive responses. J Neuroinflammation 2005, 2:24. BioMed Central Full Text
• [16]Town T: Alternative Abeta immunotherapy approaches for Alzheimer’s disease. CNS Neurol Disord Drug Targets 2009, 8:114-127.
• [17]Gordon S, Taylor PR: Monocyte and macrophage heterogeneity. Nat Rev Immunol 2005, 5:953-964.
• [18]Martinez FO, Helming L, Gordon S: Alternative activation of macrophages: an immunologic functional perspective. Annu Rev Immunol 2009, 27:451-483.
• [19]Bogdan C, Vodovotz Y, Nathan C: Macrophage deactivation by interleukin 10. J Exp Med 1991, 174:1549-1555.
• [20]Williams L, Jarai G, Smith A, Finan P: IL-10 expression profiling in human monocytes. J Leukoc Biol 2002, 72:800-809.
• [21]Li MO, Wan YY, Sanjabi S, Robertson AK, Flavell RA: Transforming growth factor-beta regulation of immune responses. Annu Rev Immunol 2006, 24:99-146.
• [22]Gregory CD, Devitt A: The macrophage and the apoptotic cell: an innate immune interaction viewed simplistically? Immunology 2004, 113:1-14.
• [23]Griffiths MR, Gasque P, Neal JW: The multiple roles of the innate immune system in the regulation of apoptosis and inflammation in the brain. J Neuropathol Exp Neurol 2009, 68:217-226.
• [24]Mosser DM, Edwards JP: Exploring the full spectrum of macrophage activation. Nat Rev Immunol 2008, 8:958-969.
• [25]Fadok VA, Bratton DL, Konowal A, Freed PW, Westcott JY, Henson PM: Macrophages that have ingested apoptotic cells in vitro inhibit proinflammatory cytokine production through autocrine/paracrine mechanisms involving TGF-beta, PGE2, and PAF. J Clin Invest 1998, 101:890-898.
• [26]Blobe GC, Schiemann WP, Lodish HF: Role of transforming growth factor beta in human disease. N Engl J Med 2000, 342:1350-1358.
• [27]Feng XH, Derynck R: Specificity and versatility in tgf-beta signaling through Smads. Annu Rev Cell Dev Biol 2005, 21:659-693.
• [28]Massague J, Gomis RR: The logic of TGFbeta signaling. FEBS Lett 2006, 580:2811-2820.
• [29]Engel ME, McDonnell MA, Law BK, Moses HL: Interdependent SMAD and JNK signaling in transforming growth factor-beta-mediated transcription. J Biol Chem 1999, 274:37413-37420.
• [30]Abe M, Harpel JG, Metz CN, Nunes I, Loskutoff DJ, Rifkin DB: An assay for transforming growth factor-beta using cells transfected with a plasminogen activator inhibitor-1 promoter-luciferase construct. Anal Biochem 1994, 216:276-284.
• [31]Oh CK, Geba GP, Molfino N: Investigational therapeutics targeting the IL-4/IL-13/STAT-6 pathway for the treatment of asthma. Eur Respir Rev 2010, 19:46-54.
• [32]Le Y, Iribarren P, Gong W, Cui Y, Zhang X, Wang JM: TGF-beta1 disrupts endotoxin signaling in microglial cells through Smad3 and MAPK pathways. J Immunol 2004, 173:962-968.
• [33]Loane DJ, Stoica BA, Pajoohesh-Ganji A, Byrnes KR, Faden AI: Activation of metabotropic glutamate receptor 5 modulates microglial reactivity and neurotoxicity by inhibiting NADPH oxidase. J Biol Chem 2009, 284:15629-15639.
• [34]Kitamura Y, Takahashi H, Matsuoka Y, Tooyama I, Kimura H, Nomura Y, Taniguchi T: In vivo induction of inducible nitric oxide synthase by microinjection with interferon-gamma and lipopolysaccharide in rat hippocampus. Glia 1996, 18:233-243.
• [35]Wynn TA: Fibrotic disease and the T(H)1/T(H)2 paradigm. Nat Rev Immunol 2004, 4:583-594.
• [36]Munder M, Eichmann K, Modolell M: Alternative metabolic states in murine macrophages reflected by the nitric oxide synthase/arginase balance: competitive regulation by CD4+ T cells correlates with Th1/Th2 phenotype. J Immunol 1998, 160:5347-5354.
• [37]Pauleau AL, Rutschman R, Lang R, Pernis A, Watowich SS, Murray PJ: Enhancer-mediated control of macrophage-specific arginase I expression. J Immunol 2004, 172:7565-7573.
• [38]Morris SM: Enzymes of arginine metabolism. J Nutr 2004, 134:2743S-2747S. discussion 2765S-2767S
• [39]Gotoh T, Sonoki T, Nagasaki A, Terada K, Takiguchi M, Mori M: Molecular cloning of cDNA for nonhepatic mitochondrial arginase (arginase II) and comparison of its induction with nitric oxide synthase in a murine macrophage-like cell line. FEBS Lett 1996, 395:119-122.
• [40]Kepka-Lenhart D, Ash DE, Morris SM: Determination of mammalian arginase activity. Methods Enzymol 2008, 440:221-230.
• [41]Loke P, Gallagher I, Nair MG, Zang X, Brombacher F, Mohrs M, Allison JP, Allen JE: Alternative activation is an innate response to injury that requires CD4+ T cells to be sustained during chronic infection. J Immunol 2007, 179:3926-3936.
• [42]Hesse M, Modolell M, La Flamme AC, Schito M, Fuentes JM, Cheever AW, Pearce EJ, Wynn TA: Differential regulation of nitric oxide synthase-2 and arginase-1 by type 1/type 2 cytokines in vivo: granulomatous pathology is shaped by the pattern of L-arginine metabolism. J Immunol 2001, 167:6533-6544.
• [43]Ma TC, Campana A, Lange PS, Lee HH, Banerjee K, Bryson JB, Mahishi L, Alam S, Giger RJ, Barnes S, Morris SM, Willis DE, Twiss JL, Filbin MT, Ratan RR: A large-scale chemical screen for regulators of the arginase 1 promoter identifies the soy isoflavone daidzeinas a clinically approved small molecule that can promote neuronal protection or regeneration via a cAMP-independent pathway. J Neurosci 2010, 30:739-748.
• [44]Chang NC, Hung SI, Hwa KY, Kato I, Chen JE, Liu CH, Chang AC: A macrophage protein, Ym1, transiently expressed during inflammation is a novel mammalian lectin. J Biol Chem 2001, 276:17497-17506.
• [45]Giannetti N, Moyse E, Ducray A, Bondier JR, Jourdan F, Propper A, Kastner A: Accumulation of Ym1/2 protein in the mouse olfactory epithelium during regeneration and aging. Neuroscience 2004, 123:907-917.
• [46]Nio J, Fujimoto W, Konno A, Kon Y, Owhashi M, Iwanaga T: Cellular expression of murine Ym1 and Ym2, chitinase family proteins, as revealed by in situ hybridization and immunohistochemistry. Histochem Cell Biol 2004, 121:473-482.
• [47]Kitowska K, Zakrzewicz D, Konigshoff M, Chrobak I, Grimminger F, Seeger W, Bulau P, Eickelberg O: Functional role and species-specific contribution of arginases in pulmonary fibrosis. Am J Physiol Lung Cell Mol Physiol 2008, 294:L34-L45.
• [48]Witte MB, Barbul A, Schick MA, Vogt N, Becker HD: Upregulation of arginase expression in wound-derived fibroblasts. J Surg Res 2002, 105:35-42.
• [49]Jiang J, George SC: TGF-{beta}2 reduces nitric oxide synthase mRNA through a ROCK-dependent pathway in airway epithelial cells. Am J Physiol Lung Cell Mol Physiol 2011, 301:L361-7.
• [50]Boutard V, Havouis R, Fouqueray B, Philippe C, Moulinoux JP, Baud L: Transforming growth factor-beta stimulates arginase activity in macrophages. Implications for the regulation of macrophage cytotoxicity. J Immunol 1995, 155:2077-2084.
• [51]Shearer JD, Richards JR, Mills CD, Caldwell MD: Differential regulation of macrophage arginine metabolism: a proposed role in wound healing. Am J Physiol 1997, 272:E181-190.
• [52]Suzumura A, Sawada M, Yamamoto H, Marunouchi T: Transforming growth factor-beta suppresses activation and proliferation of microglia in vitro. J Immunol 1993, 151:2150-2158.
• [53]Herrera-Molina R, von Bernhardi R: Transforming growth factor-beta 1 produced by hippocampal cells modulates microglial reactivity in culture. Neurobiol Dis 2005, 19:229-236.
• [54]Basu A, Krady JK, Enterline JR, Levison SW: Transforming growth factor beta1 prevents IL-1beta-induced microglial activation, whereas TNFalpha- and IL-6-stimulated activation are not antagonized. Glia 2002, 40:109-120.
• [55]Dhandapani KM, Brann DW: Transforming growth factor-beta: a neuroprotective factor in cerebral ischemia. Cell Biochem Biophys 2003, 39:13-22.
• [56]Wyss-Coray T, Masliah E, Mallory M, McConlogue L, Johnson-Wood K, Lin C, Mucke L: Amyloidogenic role of cytokine TGF-beta1 in transgenic mice and in Alzheimer’s disease. Nature 1997, 389:603-606.
• [57]Town T, Laouar Y, Pittenger C, Mori T, Szekely CA, Tan J, Duman RS, Flavell RA: Blocking TGF-beta-Smad2/3 innate immune signaling mitigates Alzheimer-like pathology. Nat Med 2008, 14:681-687.
• [58]Colton CA: Heterogeneity of microglial activation in the innate immune response in the brain. J Neuroimmune Pharmacol 2009, 4:399-418.
• [59]Lumeng CN, Bodzin JL, Saltiel AR: Obesity induces a phenotypic switch in adipose tissue macrophage polarization. J Clin Invest 2007, 117:175-184.
• [60]Lang R, Patel D, Morris JJ, Rutschman RL, Murray PJ: Shaping gene expression in activated and resting primary macrophages by IL-10. J Immunol 2002, 169:2253-2263.