In vitro system for modeling influenza A virus resistance under drug pressure

Ashley N. Brown, James J. McSharry, Qingmei Weng, Elizabeth M. Driebe, David M. Engelthaler, Kelly Sheff, Paul S Keim, Jack Nguyen, George L. Drusano

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

One of the biggest challenges in the effort to treat and contain influenza A virus infections is the emergence of resistance during treatment. It is well documented that resistance to amantadine arises rapidly during the course of treatment due to mutations in the gene coding for the M2 protein. To address this problem, it is critical to develop experimental systems that can accurately model the selection of resistance under drug pressure as seen in humans. We used the hollow-fiber infection model (HFIM) system to examine the effect of amantadine on the replication of influenza virus, A/Albany/1/98 (H3N2), grown in MDCK cells. At 24 and 48 h postinfection, virus replication was inhibited in a dose-dependent fashion. At 72 and 96 h postinfection, virus replication was no longer inhibited, suggesting the emergence of amantadine-resistant virus. Sequencing of the M2 gene revealed that mutations appeared at between 48 and 72 h of drug treatment and that the mutations were identical to those identified in the clinic for amantadine-resistant viruses (e.g., V27A, A30T, and S31N). Interestingly, we found that the type of mutation was strongly affected by the dose of the drug. The data suggest that the HFIM is a good model for influenza virus infection and resistance generation in humans. The HFIM has the advantage of being a highly controlled system where multiplicity parameters can be directly and accurately controlled and measured.

Original languageEnglish (US)
Pages (from-to)3442-3450
Number of pages9
JournalAntimicrobial Agents and Chemotherapy
Volume54
Issue number8
DOIs
StatePublished - Aug 1 2010
Externally publishedYes

Fingerprint

Amantadine
Influenza A virus
Drug Resistance
Pressure
Mutation
Virus Diseases
Virus Replication
Infection
Viruses
Madin Darby Canine Kidney Cells
Orthomyxoviridae
Pharmaceutical Preparations
Genes
In Vitro Techniques
Proteins

ASJC Scopus subject areas

  • Pharmacology (medical)
  • Pharmacology
  • Infectious Diseases

Cite this

Brown, A. N., McSharry, J. J., Weng, Q., Driebe, E. M., Engelthaler, D. M., Sheff, K., ... Drusano, G. L. (2010). In vitro system for modeling influenza A virus resistance under drug pressure. Antimicrobial Agents and Chemotherapy, 54(8), 3442-3450. https://doi.org/10.1128/AAC.01385-09

In vitro system for modeling influenza A virus resistance under drug pressure. / Brown, Ashley N.; McSharry, James J.; Weng, Qingmei; Driebe, Elizabeth M.; Engelthaler, David M.; Sheff, Kelly; Keim, Paul S; Nguyen, Jack; Drusano, George L.

In: Antimicrobial Agents and Chemotherapy, Vol. 54, No. 8, 01.08.2010, p. 3442-3450.

Research output: Contribution to journalArticle

Brown, AN, McSharry, JJ, Weng, Q, Driebe, EM, Engelthaler, DM, Sheff, K, Keim, PS, Nguyen, J & Drusano, GL 2010, 'In vitro system for modeling influenza A virus resistance under drug pressure', Antimicrobial Agents and Chemotherapy, vol. 54, no. 8, pp. 3442-3450. https://doi.org/10.1128/AAC.01385-09
Brown AN, McSharry JJ, Weng Q, Driebe EM, Engelthaler DM, Sheff K et al. In vitro system for modeling influenza A virus resistance under drug pressure. Antimicrobial Agents and Chemotherapy. 2010 Aug 1;54(8):3442-3450. https://doi.org/10.1128/AAC.01385-09
Brown, Ashley N. ; McSharry, James J. ; Weng, Qingmei ; Driebe, Elizabeth M. ; Engelthaler, David M. ; Sheff, Kelly ; Keim, Paul S ; Nguyen, Jack ; Drusano, George L. / In vitro system for modeling influenza A virus resistance under drug pressure. In: Antimicrobial Agents and Chemotherapy. 2010 ; Vol. 54, No. 8. pp. 3442-3450.
@article{ac3200d815634cbfb4a77006543a6353,
title = "In vitro system for modeling influenza A virus resistance under drug pressure",
abstract = "One of the biggest challenges in the effort to treat and contain influenza A virus infections is the emergence of resistance during treatment. It is well documented that resistance to amantadine arises rapidly during the course of treatment due to mutations in the gene coding for the M2 protein. To address this problem, it is critical to develop experimental systems that can accurately model the selection of resistance under drug pressure as seen in humans. We used the hollow-fiber infection model (HFIM) system to examine the effect of amantadine on the replication of influenza virus, A/Albany/1/98 (H3N2), grown in MDCK cells. At 24 and 48 h postinfection, virus replication was inhibited in a dose-dependent fashion. At 72 and 96 h postinfection, virus replication was no longer inhibited, suggesting the emergence of amantadine-resistant virus. Sequencing of the M2 gene revealed that mutations appeared at between 48 and 72 h of drug treatment and that the mutations were identical to those identified in the clinic for amantadine-resistant viruses (e.g., V27A, A30T, and S31N). Interestingly, we found that the type of mutation was strongly affected by the dose of the drug. The data suggest that the HFIM is a good model for influenza virus infection and resistance generation in humans. The HFIM has the advantage of being a highly controlled system where multiplicity parameters can be directly and accurately controlled and measured.",
author = "Brown, {Ashley N.} and McSharry, {James J.} and Qingmei Weng and Driebe, {Elizabeth M.} and Engelthaler, {David M.} and Kelly Sheff and Keim, {Paul S} and Jack Nguyen and Drusano, {George L.}",
year = "2010",
month = "8",
day = "1",
doi = "10.1128/AAC.01385-09",
language = "English (US)",
volume = "54",
pages = "3442--3450",
journal = "Antimicrobial Agents and Chemotherapy",
issn = "0066-4804",
publisher = "American Society for Microbiology",
number = "8",

}

TY - JOUR

T1 - In vitro system for modeling influenza A virus resistance under drug pressure

AU - Brown, Ashley N.

AU - McSharry, James J.

AU - Weng, Qingmei

AU - Driebe, Elizabeth M.

AU - Engelthaler, David M.

AU - Sheff, Kelly

AU - Keim, Paul S

AU - Nguyen, Jack

AU - Drusano, George L.

PY - 2010/8/1

Y1 - 2010/8/1

N2 - One of the biggest challenges in the effort to treat and contain influenza A virus infections is the emergence of resistance during treatment. It is well documented that resistance to amantadine arises rapidly during the course of treatment due to mutations in the gene coding for the M2 protein. To address this problem, it is critical to develop experimental systems that can accurately model the selection of resistance under drug pressure as seen in humans. We used the hollow-fiber infection model (HFIM) system to examine the effect of amantadine on the replication of influenza virus, A/Albany/1/98 (H3N2), grown in MDCK cells. At 24 and 48 h postinfection, virus replication was inhibited in a dose-dependent fashion. At 72 and 96 h postinfection, virus replication was no longer inhibited, suggesting the emergence of amantadine-resistant virus. Sequencing of the M2 gene revealed that mutations appeared at between 48 and 72 h of drug treatment and that the mutations were identical to those identified in the clinic for amantadine-resistant viruses (e.g., V27A, A30T, and S31N). Interestingly, we found that the type of mutation was strongly affected by the dose of the drug. The data suggest that the HFIM is a good model for influenza virus infection and resistance generation in humans. The HFIM has the advantage of being a highly controlled system where multiplicity parameters can be directly and accurately controlled and measured.

AB - One of the biggest challenges in the effort to treat and contain influenza A virus infections is the emergence of resistance during treatment. It is well documented that resistance to amantadine arises rapidly during the course of treatment due to mutations in the gene coding for the M2 protein. To address this problem, it is critical to develop experimental systems that can accurately model the selection of resistance under drug pressure as seen in humans. We used the hollow-fiber infection model (HFIM) system to examine the effect of amantadine on the replication of influenza virus, A/Albany/1/98 (H3N2), grown in MDCK cells. At 24 and 48 h postinfection, virus replication was inhibited in a dose-dependent fashion. At 72 and 96 h postinfection, virus replication was no longer inhibited, suggesting the emergence of amantadine-resistant virus. Sequencing of the M2 gene revealed that mutations appeared at between 48 and 72 h of drug treatment and that the mutations were identical to those identified in the clinic for amantadine-resistant viruses (e.g., V27A, A30T, and S31N). Interestingly, we found that the type of mutation was strongly affected by the dose of the drug. The data suggest that the HFIM is a good model for influenza virus infection and resistance generation in humans. The HFIM has the advantage of being a highly controlled system where multiplicity parameters can be directly and accurately controlled and measured.

UR - http://www.scopus.com/inward/record.url?scp=77955409152&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77955409152&partnerID=8YFLogxK

U2 - 10.1128/AAC.01385-09

DO - 10.1128/AAC.01385-09

M3 - Article

VL - 54

SP - 3442

EP - 3450

JO - Antimicrobial Agents and Chemotherapy

JF - Antimicrobial Agents and Chemotherapy

SN - 0066-4804

IS - 8

ER -