Abstract
The BeAn strain of Theiler’s murine encephalomyelitis virus (TMEV) induces demyelinating disease in susceptible mice comparable to human multiple sclerosis. Recent in vivo studies showed that matrix metalloproteinases (MMPs) and their inhibitors (tissue inhibitors of MMPs, TIMPs) are associated with demyelination in Theiler’s murine encephalomyelitis. The present study was performed to evaluate the in vitro MMP and TIMP expression in astrocytes and microglia following TMEV infection. Brain cell cultures from SJL/J mice were infected with the BeAn strain of TMEV and the expressions of 11 MMPs and 4 TIMPs were evaluated by reverse-transcription quantitative polymerase chain reaction (RT-qPCR) at different time points post infection (p.i.). In control astrocytes and microglia, a constitutive expression of MMP-2, -3, -9, -10, -12, -13,-14, -15, -24 and TIMP-2 to -4 was detected. In addition, TIMP-1 and MMP-11 was found in astrocytes only, and MMP-7 was absent in both cells cultures. RT-qPCR demonstrated high virus RNA copy numbers in astrocytes and a low amount in microglia. In accordance, TMEV antigen was detected in astrocytes, whereas it was below the limit of detection in microglia. MMP-3, -9, -10, -12, and -13 as well as TIMP-1 were the enzymes most prominently up-regulated in TMEV-infected astrocytes. In contrast, TMEV infection was associated with a down-regulation of MMPs and TIMPs in microglia. Conclusively, in addition to inflammatory infiltrates, TMEV-induced astrocytic MMPs might trigger a proteolysis cascade leading to an opening of the blood-brain barrier and demyelination in vivo.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Alldinger S, Gröters S, Miao Q, Fonfara, S, Kremmer, E, Baumgartner, W (2006). Roles of an extracellular matrix (ECM) receptor and ECM processing enzymes in demyelinating canine distemper encephalitis. Dtsch tierärztl Wschr 113: 151–168.
Aubert C, Chamorro M, Brahic M (1987). Identification of Theiler’s virus infected cells in the central nervous system of the mouse during demyelinating disease. Microb Pathogen 3: 319–326.
Baker AH, Edwards DR, Murphy G (2002). Metalloproteinase inhibitors: biological actions and therapeutic opportunities. J Cell Sci 115: 3719–3727.
Bottenstein JE, Sato GH (1979). Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proc Natl Acad Sci U S A 76: 514–517.
Brahic M (2002). Theiler’s virus infection of the mouse, or: of the importance of studying animal models. Virology 301: 1–5.
Brew K, Dinakarpandian D, Nagase H (2000). Tissue inhibitors of metalloproteinases: evolution, structure and function. Biochim Biophys Acta 1477: 267–283.
Carpentier PA, Williams BR (2007). Distinct roles of protein kinase R and toll-like receptor 3 in the activation of astrocytes by viral stimuli. Glia 55: 239–252.
Chandler S, Cossins J, Lury J, Wells G (1996). Macrophage metalloelastase degrades matrix and myelin proteins and processes a tumour necrosis factor-alpha fusion protein. Biochem Biophys Res Commun 228: 421–429.
Clements JM, Cossins JA, Wells GM, Corkill DJ, Helfrich K, Wood LM, Pigott R, Stabler G, Ward GA, Gearing AJ, Miller KM (1997). Matrix metalloproteinases expression during experimental autoimmune encephalomyelitis and effects of a combined matrix metalloproteinase and tumour necrosis factor-alpha inhibitor. J Neuroimmunol 74: 85–94.
Cossins JA, Clements JM, Ford J, Miller KM, Pigott R, Vos W, van der Valk P, de Groot CJ (1997). Enhanced expression of MMP-7 and MMP-9 in demyelinating multiple sclerosis lesions. Acta Neuropathol 94: 590–598.
Crocker SJ, Milner R, Pham-Mitchell N, Campbell IL (2006). Cell and agonist-specific regulation of genes for matrix metalloproteinases and their tissue inhibitors by primary glial cells. J Neurochem 98: 812–823.
Cuzner ML, Opdenakker G (1999). Plasminogen activators and matrix metalloproteases, mediators of extracellular proteolysis in inflammatory demyelination of the central nervous system. J Neuroimmunol 94: 1–14.
Dal Canto MC, Lipton HL (1982). Ultrastructural immuno-histochemical localization of virus in acute and chronic demyelinating Theiler’s virus infection. Am J Pathol 106: 20–29.
Gerhauser I, Alldinger S, Baumgartner W (2007a). Ets-1 represents a pivotal transcription factor for viral clearance, inflammation, and demyelination in a mouse model of multiple sclerosis. J Neuroimmunol 188: 86–94.
Gerhauser I, Alldinger S, Ulrich R, Baumgartner W (2005). Spatio-temporal expression of immediate early genes in the central nervous system of SJL/J mice. Int J Dev Neurosci 23: 637–649.
Gerhauser I, Ulrich R, Alldinger S, Baumgartner W (2007b). Induction of activator protein-1 and nuclear factor-κB as a prerequisite for disease development in susceptible SJL/J mice after Theiler murine encephalomyelitis. J Neuropathol Exp Neurol 66: 809–818.
Gitlin L, Barchet W, Gilfillan S, Cella M, Beutler B, Flavell RA, Diamond MS, Colonna M (2006) Essential role of mda-5 in type I IFN responses to polyriboinosinic:polyribocytidylic acid and encephalomyocarditis picornavirus. Proc Natl Acad Sci U S A 103: 8459–8464.
Giulian D, Baker TJ (1986). Characterization of ameboid microglia isolated from developing mammalian brain. J Neurosci 6: 2163–2178.
Gottschall PE, Yu X, Bing B (1995). Increased production of gelatinase B (matrix metalloproteinase-9) and interleukin-6 by activated rat microglia in culture. J Neurosci Res 42: 335–342.
Gronski TJ, Martin RL, Kobayashi DK, Walsh BC, Holman MC, Huber M, Van Wart HE, Shapiro SD (1997). Hydrolysis of a broad spectrum of extracellular matrix proteins by human macrophage elastase. J Biol Chem 272: 12189–12194.
Groters S, Alldinger S, Baumgärtner W (2005). Up-regulation of mRNA for matrix metalloproteinases-9 and -14 in advanced lesions of demyelinating canine distemper leukoencephalitis. Acta Neuropathol 110: 369–382.
Hartung HP, Kieseier BC (2000). The role of matrix metalloproteinases in autoimmune damage to the central and peripheral nervous system. J Neuroimmunol 107: 140–147.
Im HJ, Muddasani P, Natarajan V, Schmid TM, Block JA, David F, van Wijnen AJ, Loeser RF (2007). Basic fibroblast growth factor stimulates matrix metalloproteinase-13 via the molecular cross-talk between mitogen activated protein kinases and protein kinase Cδ pathways in human adult articular chondrocytes. J Biol Chem 282: 11110–11121.
Jaworski DM (2000). Differential regulation of tissue inhibitor of metalloproteinase mRNA expression in response to intracranial injury. Glia 30: 199–208.
Jelachich ML, Bramlage C, Lipton HL (1999). Differentiation of M1 myeloid precursor cells into macrophages results in binding and infection by Theiler’s murine encephalomyelitis virus and apoptosis. J Virol 73: 3227–3235.
Khasigov PZ, Podobed OV, Ktzoeva SA, Gatagonova TM, Grachev SV, Shishkin SS, Berezov TT (2001). Matrix Metalloproteinases of normal human tissues. Biochemistry 66: 130–140.
Kieseier BC, Paul R, Koedel U, Seifert T, Clements JM, Gearing AJ, Pfister HW, Hartung HP (1999). Differential expression of matrix metalloproteinases in bacterial meningitis. Brain 122: 1579–1587.
Leppert D, Lindberg RL, Kappos L, Leib SL (2001). Matrix metalloproteinases: multifunctional effectors of inflammation in multiple sclerosis and bacterial meningitis. Brain Res Rev 36: 249–257.
Lindberg RLP, De Groot CJA, Montagne L, Freitag P, van der Valk P, Kappos L, Leppert D (2001). The expression profile of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in lesions and normal appearing white matter of multiple sclerosis. Brain 124: 1743–1753.
Lipton HL (1975). Theiler’s virus infection in mice: an unusual biphasic disease process leading to demyelination. Infect Immun 11: 1147–1155.
Lipton HL, Twaddle G, Jelachich ML (1995). The predominant virus antigen burden is present in macrophages in Theiler’s murine encephalomyelitis virus-induced demyelinating disease. J Virol 69: 2525–2533.
Liuzzi GM, Latronico T, Fasabo A, Carlone G, Riccio P (2004). Interferon-beta inhibits the expression of metalloproteinases in rat glial culture implications for multiple sclerosis pathogenesis and treatment. Mult Scler 10: 290–297.
Ma Z, Qin H, Benveniste EN (2001). Transcriptional suppression of matrix metalloproteinase-9 gene expression by IFN-gamma and IFN-beta: critical role of STAT-1alpha. J Immunol 167: 5150–5159.
Markus S, Failing K, Baumgartner W (2002). Increased expression of pro-inflammatory cytokines and lack of up-regulation of anti-inflammatory cytokines in early distemper CNS lesions. J Neuroimmunol 125: 30–41.
Massova I, Kotra LP, Fridman R, Mobasery S (1998). Matrix metalloproteinases: structures, evolution, and diversification. FASEBJ 12: 1075–1095.
Matrisian LM (1990). Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet 6: 121–125.
McCarthy KD, de Vellis J (1980). Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol 85: 890–902.
McCawley LJ, Matrisian LM (2001). Matrix metalloproteinases: they’re not just for matrix anymore. Curr Opin Cell Biol 13: 534–540.
Miao Q, Baumgartner W, Failing K, Alldinger S (2003). Phase-dependent expression of matrix metalloproteinases and their inhibitors in demyelinating canine distemper encephalitis. Acta Neuropathol 106: 486–494.
Mun-Bryce S, Lukes A, Wallace J, Lukes-Marx M, Rosenberg GA (2002). Stromelysin-1 and gelatinase A are upregulated before TNF-[alpha] in LPS-stimulated neuroinflammation. Brain Res 933: 42–49.
Neuhold LA, Killar L, Zhao W, Sung MLA, Warner L, Kulik J, Turner J, Wu W, Billinghurst C, Meijers T, Poole AR, Babij P, DeGennaro LJ (2001). Postnatal expression in hyaline cartilage of constitutively active human collagenase-3 (MMP-13) induces osteoarthritis in mice. J Clin Invest 107: 35–44.
Oleszak EL, Chang JR, Friedman H, Katsetos CD, Platsoucas CD (2004). Theiler’s virus infection: a model for multiple sclerosis. Clin Microbiol Rev 17: 174–207.
Pagenstecher A, Stalder AK, Kincaid CL, Shapiro SD, Campbell IL (1998). Differential expression of matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase genes in the mouse central nervous system in normal and inflammatory states. Am J Pathol 152: 729–741.
Palma JP, Kim BS (2004). The scope and activation mechanisms of chemokine gene expression in primary astrocytes following infection with Theiler’s virus. J Neuroimmunol 149: 121–129.
Palma JP, Kwon D, Clipstone NA, Kim BS (2003). Infection with Theiler’s murine encephalomyelitis virus directly induces proinflammatory cytokines in primary astrocytes via NF-kappaB activation: potential role for the initiation of demyelinating disease. J Virol 77: 6322–6331.
Racaniello VR (2001). Picornaviridae: the viruses and their replication. In: Field’s virology. Knipe, DM, Howley, PM, Griffin, DE, Lamb, RA, Martin, MA, Roizman, B, Strauss, SE (eds). Philadelphia: Lippincott Williams & Wilkins pp 685–722.
Ries C, Petrides PE (1995). Cytokine regulation of matrix metalloproteinase activity and its regulatory dysfunction in disease. Biol Chem Hoppe-Seyler 376: 345–355.
Rosenberg GA (2001). Matrix metalloproteinases in multiple sclerosis: is it time for a treatment trial? Ann Neurol 50: 431–433.
Rosenberg GA (2002). Matrix metalloproteinases in neuroinflammation. Glia 39: 279–291.
Rosenberg GA (2005). Matrix metalloproteinases biomarkers in multiple sclerosis. Lancet 365: 1291–1293.
Rosenberg GA, Estrada, EY, Dencoff, JE, Stetler-Stevenson, WG (1995). Tumor necrosis factor-alpha-induced gelatinase B causes delayed opening of the blood-brain barrier: an expanded therapeutic window. Brain Res 703: 151–155.
Rozhon EJ, Kratochvil JD, Lipton HL (1983). Analysis of genetic variation in Theiler’s virus persistent infection in the mouse central nervous system. Virology 128: 16–32.
Rubio N, Capa L (1993). Differential IL-1 synthesis by astrocytes from Theiler’s murine encephalomyelitis virus-susceptible and -resistant strains of mice. Cell Immunol 149: 237–247.
Rubio N, Martin-Clemente B (1999). Theiler’s murine encephalomyelitis virus infection induces early expression of c-fos in astrocytes. Virology 258: 21–29.
Rubio N, Sanz-Rodriguez F, Lipton HL (2006). Theiler’s virus induces the MIP-2 chemokine (CXCL2) in astrocytes from genetically susceptible but not from resistant mouse strains. Cell Immunol 239: 31–40.
Rudolph-Owen LA, Hulboy DL, Wilson CL, Mudgett J, Matrisian M (1997). Coordinate expression of matrix metalloproteinase family members in the uterus of normal, matrilysin-deficient, and stromelysin-1-deficient mice. Endocrinology 138: 4902–4911.
So EY, Kang MH, Kim BS (2006). Induction of chemokine and cytokine genes in astrocytes following infection with Theiler’s murine encephalomyelitis virus is mediated by the Toll-like receptor 3. Glia 53: 858–867.
Stamenkovic I (2003). Extracellular matrix remodeling: the role of matrix metalloproteinases. J Pathol 200: 448–464.
Stohlman SA, Hinton DR (2001). Viral induced demyelination. Brain Pathol 11: 92–106.
Teesalu T, Hinkkanen AE, Vaheri A (2001). Coordinated induction of extracellular proteolysis systems during experimental autoimmune encephalomyelitis in mice. Am J Pathol 159: 2227–2237.
Theiler M (1934). Spontaneous encephalomyelitis of mice—a new virus disease. Science 80: 122–123.
Thorns V, Walter GF, Thorns C (2003). Expression of MMP-2, MMP-7, MMP-9, MMP-10 and MMP-11 in human astrocytic and oligodendroglial gliomas. Anticancer Res 23: 3937–3944.
Toft-Hansen H, Nuttall RK, Edwards DR, Owens T (2004). Key metalloproteinases are expressed by specific cell types in experimental autoimmune encephalomyelitis. J Immunol 173: 5209–5218.
Tsunoda I, Iwasaki Y, Terunuma H, Sako K, Ohara Y (1996). A comparative study of acute and chronic diseases induced by two subgroups of Theiler’s murine encephalomyelitis virus. Acta Neuropathol 91: 595–602.
Ulrich R, Baumgartner W, Gerhauser I, Seeliger F, Haist V, Deschl U, Alldinger S (2006). MMP-12, MMP-3, and TIMP-1 are markedly upregulated in chronic demyelinating Theiler murine encephalomyelitis. J Neuropathol Exp Neurol 65: 783–793.
Ulrich R, Gerhauser I, Seeliger F, Baumgartner W, Alldinger S (2005). Matrix metalloproteinases and their inhibitors in the developing mouse brain and spinal cord: a reverse transcription quantitative polymerase chain reaction study. Dev Neurosci 27: 408–418.
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002). Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol, 3, published online: http://genomebiology.com/ 2002/3/7/research/0034.
Vos CM, van Haastert ES, de Groot CJ, van der Valk P, de Vries HE (2003). Matrix metalloproteinase-12 is expressed in phagocytotic macrophages in active multiple sclerosis lesions. J Neuroimmunol 138: 106–114.
Welsh CJ, Tonks P, Borrow P, Nash AA (1990). Theiler’s virus: an experimental model of virus-induced demyelination. Autoimmunity 6: 105–112.
Woessner JF (1991). Matrix metalloproteinases and their inhibitors in connective tissue remodeling. FASEB J 5: 2145–2153.
Yong VW, Krekoski CA, Forsyth PA, Bell R, Edwards DR (1998). Matrix metalloproteinases and diseases of the CNS. Trends Neurosci 21: 75–80.
Zheng L, Calenoff MA, Dal Canto MC (2001). Astrocytes, not microglia, are the main cells responsible for viral persistence in Theiler’s murine encephalomyelitis virus infection leading to demyelination. J Neuroimmunol 118: 256–267.
Zhou J, Marten NW, Bergmann CC, Macklin WB, Hinton DR, Stohlman SA (2005). Expression of matrix metalloproteinases and their tissue inhibitor during viral encephalitis. J Virol 79: 4764–4773.
Zhou J, Stohlman SA, Atkinson R, Hinton DR, Marten NW (2002). Matrix metalloproteinase expression correlates with virulence following neurotropic mouse hepatitis virus infection. J Virol 76: 7374–7384.
Zoecklein LJ, Pavelko KD, Gamez J, Papke L, McGavern DB, Ure DR, Njenga MK, Johnson AJ, Nakane S, Rodriguez M (2003). Direct comparison of demyelinating disease induced by the Daniel’s strain and BeAn strain of Theiler’s murine encephalomyelitis virus. Brain Pathol 13: 291–308.
Author information
Authors and Affiliations
Corresponding author
Additional information
Jirawat Kumnok and Reiner Ulrich contributed equally to this work.
This article is in memory of Jirawat Kumnok, a young outstanding scientist who passed away far too early.
Rights and permissions
About this article
Cite this article
Kumnok, J., Ulrich, R., Wewetzer, K. et al. Differential transcription of matrix-metalloproteinase genes in primary mouse astrocytes and microglia infected with Theiler’s murine encephalomyelitis virus. Journal of NeuroVirology 14, 205–217 (2008). https://doi.org/10.1080/13550280802008305
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1080/13550280802008305