title: The replication of a mouse adapted SARS-CoV in a mouse cell line stably expressing the murine SARS-CoV receptor mACE2 efficiently induces the expression of proinflammatory cytokines.
abstract: Infection of conventional mice with a mouse adapted (MA15) severe acute respiratory syndrome (SARS) coronavirus (CoV) reproduces many aspects of human SARS such as pathological changes in lung, viremia, neutrophilia, and lethality. However, established mouse cell lines highly susceptible to mouse-adapted SARS-CoV infection are not available. In this work, efficiently transfectable mouse cell lines stably expressing the murine SARS-CoV receptor angiotensin converting enzyme 2 (ACE2) have been generated. These cells yielded high SARS-CoV-MA15 titers and also served as excellent tools for plaque assays. In addition, in these cell lines, SARS-CoV-MA15 induced the expression of proinflammatory cytokines and IFN-β, mimicking what has been observed in experimental animal models infected with SARS-CoV and SARS patients. These cell lines are valuable tools to perform in vitro studies in a mouse cell system that reflects the species used for in vivo studies of SARS-CoV-MA15 pathogenesis.
authors.affiliation: Department of Molecular and Cell Biology, Centro Nacional de Biotecnología (CNB-CSIC), Darwin 3, Campus Universidad Autónoma de Madrid, 28049 Madrid, Spain.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Regla-Nava
authors.forename: Jose M
authors.initials: JM
authors.lastname: Jimenez-Guardeño
authors.forename: Jose L
authors.initials: JL
authors.lastname: Nieto-Torres
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Gallagher
authors.forename: Luis
authors.initials: L
authors.lastname: Enjuanes
authors.forename: Marta L
authors.initials: ML
authors.lastname: DeDiego
mesh_terms: D000220:Adaptation, Biological; D000085962:Angiotensin-Converting Enzyme 2; D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis*; D015870:Gene Expression; D051379:Mice; D008807:Mice, Inbred BALB C; D007703:Peptidyl-Dipeptidase A / Q000235:genetics / Q000378:metabolism*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000254:growth & development* / Q000276:immunology; D019562:Viral Load; D010948:Viral Plaque Assay
chemical_list: D016207:Cytokines; D007703:Peptidyl-Dipeptidase A; C000705307:ACE2 protein, human; C000705328:Ace2 protein, mouse; D000085962:Angiotensin-Converting Enzyme 2
keywords: Coronavirus; Mouse adapted; Proinflammatory cytokines; SARS; SARS-CoV receptor ACE2; Stably transformed murine cells
references.citation: Virology. 2006 Jul 20;351(1):180-7
references.pmid: 16647731
references.citation: Infect Immun. 2004 Aug;72(8):4410-5
references.pmid: 15271897
references.citation: J Biol Chem. 2009 Jun 12;284(24):16202-16209
references.pmid: 19380580
references.citation: Jpn J Exp Med. 1967 Oct;37(5):461-74
references.pmid: 4301953
references.citation: Science. 2003 May 30;300(5624):1394-9
references.pmid: 12730500
references.citation: J Virol. 2007 Nov;81(21):11620-33
references.pmid: 17715225
references.citation: Lancet. 2003 Jul 26;362(9380):263-70
references.pmid: 12892955
references.citation: BMJ. 2003 Jun 21;326(7403):1358-62
references.pmid: 12816821
references.citation: J Virol. 2007 Aug;81(16):8692-706
references.pmid: 17537853
references.citation: J Biol Chem. 2007 Nov 2;282(44):32208-21
references.pmid: 17761676
references.citation: Am J Respir Crit Care Med. 2005 Apr 15;171(8):850-7
references.pmid: 15657466
references.citation: J Virol. 2008 Sep;82(17):8721-32
references.pmid: 18579604
references.citation: N Engl J Med. 2003 May 15;348(20):1953-66
references.pmid: 12690092
references.citation: J Virol. 2011 May;85(9):4234-45
references.pmid: 21325418
references.citation: J Med Virol. 2005 Feb;75(2):185-94
references.pmid: 15602737
references.citation: Di Yi Jun Yi Da Xue Xue Bao. 2003 Aug;23(8):770-3
references.pmid: 12919893
references.citation: Virus Res. 2008 Apr;133(1):20-32
references.pmid: 17499378
references.citation: Proc Natl Acad Sci U S A. 2000 May 9;97(10):5516-21
references.pmid: 10805807
references.citation: Virology. 2001 Jan 20;279(2):371-4
references.pmid: 11162792
references.citation: J Virol. 2008 May;82(9):4471-9
references.pmid: 18305050
references.citation: J Biol Chem. 2004 Jan 30;279(5):3197-201
references.pmid: 14670965
references.citation: Nature. 2003 Nov 27;426(6965):450-4
references.pmid: 14647384
references.citation: J Med Virol. 2004 Sep;74(1):1-7
references.pmid: 15258961
references.citation: J Virol. 2005 Mar;79(6):3846-50
references.pmid: 15731278
references.citation: PLoS Biol. 2008 Sep 16;6(9):e226
references.pmid: 18798692
references.citation: J Virol. 2013 Jun;87(12):6551-9
references.pmid: 23576515
references.citation: Virus Res. 2008 Apr;133(1):101-12
references.pmid: 17451827
references.citation: Proc Natl Acad Sci U S A. 2004 Nov 2;101(44):15748-53
references.pmid: 15496474
references.citation: N Engl J Med. 2003 May 15;348(20):1967-76
references.pmid: 12690091
references.citation: J Gen Virol. 2000 Apr;81(Pt 4):853-79
references.pmid: 10725411
references.citation: PLoS Pathog. 2010 Feb 05;6(2):e1000756
references.pmid: 20140198
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 16;105(50):19944-9
references.pmid: 19036930
references.citation: DNA Cell Biol. 2005 Aug;24(8):496-502
references.pmid: 16101347
references.citation: Virology. 2009 Dec 20;395(2):210-22
references.pmid: 19853271
references.citation: J Virol. 2002 Apr;76(8):3697-708
references.pmid: 11907209
references.citation: J Virol. 2007 Feb;81(4):1701-13
references.pmid: 17108030
references.citation: J Med Virol. 2004 Jul;73(3):323-31
references.pmid: 15170624
references.citation: Am J Pathol. 2008 Jun;172(6):1625-37
references.pmid: 18467696
references.citation: Pathology. 2003 Dec;35(6):526-31
references.pmid: 14660106
references.citation: J Virol. 2007 Jan;81(2):548-57
references.pmid: 17108024
references.citation: J Virol. 2008 Oct;82(19):9465-76
references.pmid: 18632870
references.citation: J Virol. 2007 Jan;81(2):813-21
references.pmid: 17079315
references.citation: Clin Chem. 2005 Dec;51(12):2333-40
references.pmid: 16195357
references.citation: PLoS Pathog. 2007 Aug 10;3(8):e112
references.pmid: 17696609
references.citation: PLoS Pathog. 2007 Jan;3(1):e5
references.pmid: 17222058
references.citation: PLoS Pathog. 2011 Oct;7(10):e1002315
references.pmid: 22028656
references.citation: Nature. 2003 May 15;423(6937):240
references.pmid: 12748632
references.citation: Cell Mol Life Sci. 2004 Aug;61(16):2100-12
references.pmid: 15316659
references.citation: Proc Natl Acad Sci U S A. 2005 Sep 27;102(39):14040-5
references.pmid: 16169905
references.citation: Science. 2005 Sep 16;309(5742):1864-8
references.pmid: 16166518
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Arch Virol. 2005 May;150(5):1023-31
references.pmid: 15645376
references.citation: Trends Microbiol. 2006 Jul;14(7):299-303
references.pmid: 16759866
references.citation: J Gen Virol. 2003 Dec;84(Pt 12):3291-3303
references.pmid: 14645910
references.citation: J Virol. 2006 Nov;80(21):10900-6
references.pmid: 16928748
references.citation: J Virol. 2009 Jul;83(14):7062-74
references.pmid: 19420084
references.citation: Science. 2003 May 30;300(5624):1399-404
references.pmid: 12730501
references.citation: J Virol. 2004 Oct;78(20):11429-33
references.pmid: 15452268
references.citation: Lancet. 2003 Apr 19;361(9366):1319-25
references.pmid: 12711465
references.citation: PLoS One. 2012;7(2):e30802
references.pmid: 22312431
references.citation: Comp Med. 2007 Oct;57(5):450-9
references.pmid: 17974127
references.citation: J Virol. 2007 Sep;81(18):9812-24
references.pmid: 17596301
references.citation: BMC Immunol. 2005 Jan 18;6:2
references.pmid: 15655079
references.citation: J Virol. 2008 Mar;82(5):2274-85
references.pmid: 18094188