title: Molecular determinants of severe acute respiratory syndrome coronavirus pathogenesis and virulence in young and aged mouse models of human disease.
abstract: SARS coronavirus (SARS-CoV) causes severe acute respiratory tract disease characterized by diffuse alveolar damage and hyaline membrane formation. This pathology often progresses to acute respiratory distress (such as acute respiratory distress syndrome [ARDS]) and atypical pneumonia in humans, with characteristic age-related mortality rates approaching 50% or more in immunosenescent populations. The molecular basis for the extreme virulence of SARS-CoV remains elusive. Since young and aged (1-year-old) mice do not develop severe clinical disease following infection with wild-type SARS-CoV, a mouse-adapted strain of SARS-CoV (called MA15) was developed and was shown to cause lethal infection in these animals. To understand the genetic contributions to the increased pathogenesis of MA15 in rodents, we used reverse genetics and evaluated the virulence of panels of derivative viruses encoding various combinations of mouse-adapted mutations. We found that mutations in the viral spike (S) glycoprotein and, to a much less rigorous extent, in the nsp9 nonstructural protein, were primarily associated with the acquisition of virulence in young animals. The mutations in S likely increase recognition of the mouse angiotensin-converting enzyme 2 (ACE2) receptor not only in MA15 but also in two additional, independently isolated mouse-adapted SARS-CoVs. In contrast to the findings for young animals, mutations to revert to the wild-type sequence in nsp9 and the S glycoprotein were not sufficient to significantly attenuate the virus compared to other combinations of mouse-adapted mutations in 12-month-old mice. This panel of SARS-CoVs provides novel reagents that we have used to further our understanding of differential, age-related pathogenic mechanisms in mouse models of human disease.
authors.affiliation: Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland, USA. mfrieman@som.umaryland.edu
authors.forename: Matthew
authors.initials: M
authors.lastname: Frieman
authors.forename: Boyd
authors.initials: B
authors.lastname: Yount
authors.forename: Sudhakar
authors.initials: S
authors.lastname: Agnihothram
authors.forename: Carly
authors.initials: C
authors.lastname: Page
authors.forename: Eric
authors.initials: E
authors.lastname: Donaldson
authors.forename: Anjeanette
authors.initials: A
authors.lastname: Roberts
authors.forename: Leatrice
authors.initials: L
authors.lastname: Vogel
authors.forename: Becky
authors.initials: B
authors.lastname: Woodruff
authors.forename: Diana
authors.initials: D
authors.lastname: Scorpio
authors.forename: Kanta
authors.initials: K
authors.lastname: Subbarao
authors.forename: Ralph S
authors.initials: RS
authors.lastname: Baric
mesh_terms: D000367:Age Factors; D000818:Animals; D002460:Cell Line; D004195:Disease Models, Animal; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D059386:Reverse Genetics; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000472:pathogenicity*; D045169:Severe Acute Respiratory Syndrome / Q000401:mortality / Q000473:pathology / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014774:Virulence
chemical_list: D014764:Viral Proteins
references.citation: Biochem Biophys Res Commun. 2003 Dec 26;312(4):1159-64
references.pmid: 14651994
references.citation: J Virol. 2007 Nov;81(22):12135-44
references.pmid: 17804504
references.citation: Virology. 2009 Dec 20;395(2):210-22
references.pmid: 19853271
references.citation: PLoS Pathog. 2006 Jul;2(7):e73
references.pmid: 16848640
references.citation: Proc Natl Acad Sci U S A. 2003 Oct 28;100(22):12995-3000
references.pmid: 14569023
references.citation: Science. 2003 May 30;300(5624):1394-9
references.pmid: 12730500
references.citation: Proc Natl Acad Sci U S A. 2005 Feb 15;102(7):2430-5
references.pmid: 15695582
references.citation: Lab Anim Sci. 1990 Sep;40(5):481-5
references.pmid: 2170746
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 29;94(9):4659-63
references.pmid: 9114047
references.citation: J Virol. 2008 Jun;82(11):5137-44
references.pmid: 18367528
references.citation: Can J Infect Dis Med Microbiol. 2006 Nov;17(6):330-6
references.pmid: 18382647
references.citation: J Gen Virol. 1997 Dec;78 ( Pt 12):3207-15
references.pmid: 9400971
references.citation: J Virol. 2004 Oct;78(19):10628-35
references.pmid: 15367630
references.citation: J Virol. 2008 Sep;82(17):8721-32
references.pmid: 18579604
references.citation: J Gen Virol. 2001 Jun;82(Pt 6):1365-1373
references.pmid: 11369881
references.citation: N Engl J Med. 2003 May 15;348(20):1953-66
references.pmid: 12690092
references.citation: PLoS Med. 2006 Feb;3(2):e27
references.pmid: 16379499
references.citation: J Virol. 2004 Jun;78(11):5619-32
references.pmid: 15140959
references.citation: J Virol. 1987 Sep;61(9):2917-20
references.pmid: 3039178
references.citation: Adv Exp Med Biol. 2006;581:285-8
references.pmid: 17037544
references.citation: Virus Res. 2008 Apr;133(1):20-32
references.pmid: 17499378
references.citation: PLoS Pathog. 2008 Dec;4(12):e1000240
references.pmid: 19079579
references.citation: Proc Natl Acad Sci U S A. 2004 Mar 16;101(11):3792-6
references.pmid: 15007178
references.citation: Adv Exp Med Biol. 2001;494:101-7
references.pmid: 11774452
references.citation: Science. 2005 Sep 16;309(5742):1822-3
references.pmid: 16166506
references.citation: PLoS Pathog. 2010 May 06;6(5):e1000896
references.pmid: 20463816
references.citation: Science. 2003 Oct 10;302(5643):276-8
references.pmid: 12958366
references.citation: Science. 2004 Mar 12;303(5664):1666-9
references.pmid: 14752165
references.citation: Proc Natl Acad Sci U S A. 2000 Sep 12;97(19):10578-83
references.pmid: 10973478
references.citation: J Virol. 2005 Mar;79(6):3391-400
references.pmid: 15731233
references.citation: J Virol. 2008 Jul;82(14):6984-91
references.pmid: 18448527
references.citation: J Virol. 2004 Dec;78(24):13600-12
references.pmid: 15564471
references.citation: Lancet. 2003 May 24;361(9371):1767-72
references.pmid: 12781535
references.citation: Curr Opin Pharmacol. 2006 Jun;6(3):271-6
references.pmid: 16581295
references.citation: Hum Pathol. 2006 Apr;37(4):381-90
references.pmid: 16564911
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: RNA Biol. 2011 Mar-Apr;8(2):270-9
references.pmid: 21593585
references.citation: J Virol. 1996 Oct;70(10):6673-81
references.pmid: 8794303
references.citation: J Virol. 2009 Dec;83(24):12998-3008
references.pmid: 19828617
references.citation: N Engl J Med. 2003 May 15;348(20):1967-76
references.pmid: 12690091
references.citation: Science. 1949 Aug 19;110(2851):184
references.pmid: 17779824
references.citation: EMBO J. 2005 Apr 20;24(8):1634-43
references.pmid: 15791205
references.citation: Cell. 1990 Oct 19;63(2):353-62
references.pmid: 2170026
references.citation: J Gen Virol. 1980 Jan;46(1):1-13
references.pmid: 6766176
references.citation: Structure. 2004 Feb;12(2):341-53
references.pmid: 14962394
references.citation: J Virol. 2004 Sep;78(18):9977-86
references.pmid: 15331731
references.citation: PLoS Pathog. 2009 Oct;5(10):e1000636
references.pmid: 19851468
references.citation: Nucleic Acids Res. 2010 Nov;38(21):7626-36
references.pmid: 20671029
references.citation: J Virol. 2007 Jan;81(2):813-21
references.pmid: 17079315
references.citation: Histopathology. 2004 Aug;45(2):119-24
references.pmid: 15279629
references.citation: Virology. 2004 Feb 20;319(2):262-73
references.pmid: 14980486
references.citation: J Virol. 2011 Jan;85(1):217-30
references.pmid: 20980507
references.citation: J Virol. 2010 Nov;84(21):11297-309
references.pmid: 20702617
references.citation: Curr Opin Immunol. 2005 Aug;17(4):404-10
references.pmid: 15950449
references.citation: J Virol. 2005 Dec;79(24):15189-98
references.pmid: 16306590
references.citation: PLoS Pathog. 2007 Jan;3(1):e5
references.pmid: 17222058
references.citation: J Virol. 2004 Mar;78(6):2701-10
references.pmid: 14990690
references.citation: Proc Natl Acad Sci U S A. 2005 Jun 7;102(23):8275-80
references.pmid: 15919828
references.citation: Science. 2005 Sep 16;309(5742):1864-8
references.pmid: 16166518
references.citation: J Virol. 2005 May;79(9):5833-8
references.pmid: 15827197
references.citation: J Virol. 2009 Jul;83(13):6689-705
references.pmid: 19369340
references.citation: PLoS Pathog. 2010 Apr 08;6(4):e1000849
references.pmid: 20386712
references.citation: Cell. 2003 Jun 13;113(6):701-2
references.pmid: 12809601
references.citation: Am J Trop Med Hyg. 1985 Mar;34(2):388-95
references.pmid: 2858987
references.citation: J Virol. 2007 Jul;81(14):7410-23
references.pmid: 17507479
references.citation: N Engl J Med. 2003 Dec 18;349(25):2431-41
references.pmid: 14681510
references.citation: Lancet. 2003 May 24;361(9371):1773-8
references.pmid: 12781536
references.citation: Infect Immun. 1981 Jan;31(1):334-8
references.pmid: 6111538
references.citation: Hong Kong Med J. 2010;16(5 Suppl 4):29-33
references.pmid: 20864745
references.citation: J Virol. 2009 Jul;83(14):7062-74
references.pmid: 19420084
references.citation: J Virol. 2004 May;78(9):4552-60
references.pmid: 15078936
references.citation: J Immunol. 2004 Sep 15;173(6):4030-9
references.pmid: 15356152
references.citation: Emerg Infect Dis. 2004 Sep;10(9):1550-7
references.pmid: 15498155
references.citation: J Virol. 2008 Mar;82(5):2274-85
references.pmid: 18094188