title: Analysis of oseltamivir resistance substitutions in influenza virus glycoprotein neuraminidase using a lentivirus-based surrogate assay system.
abstract: Influenza A virus poses a great threat to global health, and oseltamivir (trade marked as Tamiflu), which targets influenza surface glycoprotein neuraminidase (NA), is used clinically as a major anti-influenza treatment. However, certain substitutions in NA can render an influenza virus resistant to this drug. In this study, using a lentiviral pseudotyping system, which alleviates the safety concerns of studying highly pathogenic influenza viruses such as avian influenza H5N1, that utilizes influenza surface glycoproteins (hemagglutinin or HA, and NA) and an HIV-core combined with a luciferase reporter gene as a surrogate assay, we first assessed the functionality of NA by measuring pseudovirion release in the absence or presence of oseltamivir. We demonstrated that oseltamivir displays a dose-dependent inhibition on NA activity. In contrast, a mutant NA (H274Y) is more resistant to oseltamivir treatment. In addition, the effects of several previously reported substitution NA mutants were examined as well. Our results demonstrate that this lentivirus-based pseudotyping system provides a quick, safe, and effective way to assess resistance to neuraminidase inhibitors. And we believe that as new mutations appear in influenza isolates, their impact on the effectiveness of current and future anti-NA can be quickly and reliably evaluated by this assay.
authors: Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicago, Chicago, IL 60612, USA.
authors: Jennifer
authors: J
authors: Tisoncik-Go
authors: Katie S
authors: KS
authors: Cordero
authors: Lijun
authors: L
authors: Rong
mesh_terms: D002460:Cell Line; D024882:Drug Resistance, Viral / Q000235:genetics; D006384:Hemagglutination; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D009439:Neuraminidase / Q000235:genetics / Q000378:metabolism*; D009975:Orthomyxoviridae / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D053139:Oseltamivir / Q000031:analogs & derivatives / Q000494:pharmacology*
chemical_list: D053139:Oseltamivir; D009439:Neuraminidase; C535162:oseltamivir carboxylate
references: Virology. 1995 Oct 1;212(2):323-30
references: 7571401
references: N Engl J Med. 2005 Dec 22;353(25):2667-72
references: 16371632
references: J Virol. 2007 Nov;81(22):12418-26
references: 17855542
references: J Med Chem. 2005 Jun 16;48(12):3980-90
references: 15943472
references: Nature. 2006 Sep 7;443(7107):45-9
references: 16915235
references: Science. 2005 Aug 19;309(5738):1206
references: 16000410
references: Antimicrob Agents Chemother. 2008 Oct;52(10):3484-91
references: 18694948
references: J Control Release. 1999 Aug 5;60(2-3):149-60
references: 10425321
references: Nature. 2005 Oct 20;437(7062):1108
references: 16228009
references: Biochemistry. 2003 Jan 28;42(3):718-27
references: 12534284
references: Virology. 1997 Sep 15;236(1):66-75
references: 9299618
references: J Virol. 1995 Nov;69(11):6705-11
references: 7474080
references: Antiviral Res. 2007 Mar;73(3):228-31
references: 17112602
references: Antimicrob Agents Chemother. 2005 Oct;49(10):4075-84
references: 16189083
references: Antiviral Res. 2008 Feb;77(2):163-6
references: 17919743
references: Anal Biochem. 1979 Apr 15;94(2):287-96
references: 464297
references: Antimicrob Agents Chemother. 2001 Dec;45(12):3403-8
references: 11709315
references: Pediatr Infect Dis J. 2001 Feb;20(2):127-33
references: 11224828
references: Clin Infect Dis. 2006 Dec 15;43(12):1555-61
references: 17109288
references: Antiviral Res. 2002 May;54(2):99-111
references: 12062395
references: N Engl J Med. 2003 Feb 27;348(9):867-8
references: 12606750
references: Nature. 1993 Jun 3;363(6428):418-23
references: 8502295
references: Antimicrob Agents Chemother. 1998 Dec;42(12):3234-41
references: 9835519
references: J Virol. 2006 Sep;80(17):8787-95
references: 16912325
references: Antimicrob Agents Chemother. 2003 Jul;47(7):2264-72
references: 12821478
references: JAMA. 2007 Apr 4;297(13):1435-42
references: 17405969
references: Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):9852-6
references: 8234325
references: J Med Chem. 2000 Sep 21;43(19):3482-6
references: 11000002
references: J Virol. 1997 May;71(5):3385-90
references: 9094607
references: Antimicrob Agents Chemother. 1995 Nov;39(11):2583-4
references: 8585752
references: Nat Struct Biol. 2000 Nov;7(11):1068-74
references: 11062565
references: Antimicrob Agents Chemother. 2002 Apr;46(4):1014-21
references: 11897583
references: Antimicrob Agents Chemother. 1993 Jul;37(7):1473-9
references: 8363379
references: J Infect Dis. 2001 Feb 15;183(4):523-31
references: 11170976
references: J Org Chem. 2002 Aug 9;67(16):5445-53
references: 12153241
references: Lancet. 2004 Aug 28-Sep 3;364(9436):759-65
references: 15337401
references: J Infect Dis. 2006 Mar 15;193(6):760-4
references: 16479508
references: J Mol Biol. 1993 Mar 5;230(1):345-8
references: 8450546
references: J Clin Virol. 2008 Jan;41(1):20-4
references: 18065262
references: J Infect Dis. 2004 Nov 1;190(9):1627-30
references: 15478068
references: Nature. 2008 Jun 26;453(7199):1258-61
references: 18480754