Characterization of Drug-Resistant Influenza A(H7N9) Variants Isolated From an Oseltamivir-Treated Patient in Taiwan

Marjuki, Henju; Mishin, Vasiliy P.; Chesnokov, Anton; Jones, Joyce; Cruz, Juan A. De La; Sleeman, Katrina; Tamura, Daisuke; Nguyen, Ha; Wu, Ho-Sheng; Chang, Feng–Yee; Liu, Ming‐Tsan; Fry, Alicia M.; Cox, Nancy J.; Villanueva, Julie; Davis, C. Todd; Gubareva, Larisa V.

doi:10.1093/infdis/jiu447

Cited by 73 publications

(69 citation statements)

References 43 publications

(55 reference statements)

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“…Currently, oseltamivir is the first choice treatment, which has a significant effect on influenza A virus replication and inflammatory cytokine production, and can obviously improve the survival rate. However, an increasing number of drug‐resistant viruses have been discovered recently owing to the enormous genetic variation of influenza virus 31, 32, 33, 34, 35, 36, 37. Because of the amplified inflammatory response after H5N1 infection, anti‐inflammatory drugs have been considered to treat humans exposed to the influenza A virus.…”

Section: Discussionmentioning

confidence: 99%

The therapeutic effects of sodium cromoglycate against influenza A virus H5N1 in mice

Han

Wei

Zhang

et al. 2015

Influenza Resp Viruses

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ObjectivesTo identify the protective role of sodium cromoglycate in mice during influenza virus infection.DesignH5N1 virus‐infected mice were treated with the mast cell stabilizer sodium cromoglycate (SCG) to investigate its therapeutic effect.SampleThe nose, trachea and lungs from mice were collected.Main outcome measuresVirus replication and host responses were determined by plaque assay, quantitative PCR, immunohistochemistry, and histology.ResultsSCG‐treated mice survived better than did PBS‐treated mice after H5N1 virus infection. Mild pathological changes with fewer inflammatory cell infiltration and fewer virus antigens were observed in the nose, trachea, and lungs of SCG‐treated mice on days 3 and 5 post‐infection. However, no significant changes in viral load in the lungs were detected between SCG‐ and PBS‐treated mice. Furthermore, significantly decreased expression of interleukin‐6, tumor necrosis factor‐a, Toll‐like receptor 3, and TIR‐domain‐containing adapter‐inducing interferon‐b was detected in the lungs of SCG‐treated mice, and no higher expression of interferon‐c was detected.ConclusionThese results suggest that SCG has therapeutic roles in H5N1 virus‐infected mice by alleviating the inflammatory response rather than inhibition of viral replication in the lungs.

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Section: Discussionmentioning

confidence: 99%

The therapeutic effects of sodium cromoglycate against influenza A virus H5N1 in mice

Han

Wei

Zhang

et al. 2015

Influenza Resp Viruses

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“…A number of oseltamivir-resistant clinical A(H7N9) isolates have been identified. Mutations in the NA that are associated with antiviral resistance in these viruses include E119V, I222R or I222K, A246T, and R292K (8,29,31,32). We rescued H6N9 AH/13 viruses that contained 119V, 222R, 246T, or 292K in the NA and tested the ability of MAbs 1E8, 2F6, 11B2, and 10F4 to inhibit their NA activity.…”

Section: Preparation Of N9 Mabs a Panel Of 19mentioning

confidence: 99%

Comparison of the Efficacy of N9 Neuraminidase-Specific Monoclonal Antibodies against Influenza A(H7N9) Virus Infection

Wan

Gao

et al. 2018

J Virol

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The fifth wave of A(H7N9) virus infection in China from 2016 to 2017 caused great concern due to the large number of individuals infected, the isolation of drug-resistant viruses, and the emergence of highly pathogenic strains. Antibodies against neuraminidase (NA) provide added benefit to hemagglutinin-specific immunity and may be important contributors to the effectiveness of A(H7N9) vaccines. We generated a panel of mouse monoclonal antibodies (MAbs) to identify antigenic domains on NA of the novel A(H7N9) virus and compared their functional properties. The loop formed in the region of residue 250 (250 loop) and the domain formed by the loops containing residues 370, 400, and 430 were identified as major antigenic regions. MAbs 1E8, 2F6, 10F4, and 11B2, which recognize these two antigenic domains, were characterized in depth. These four MAbs differ in their abilities to inhibit cleavage of small and large substrates (methyl-umbelliferylacetyl neuraminic acid [MU-NANA] and fetuin, respectively) in NA inhibition assays. 1E8 and 11B2 did not inhibit NA cleavage of either MU-NANA or fetuin, and 2F6 inhibited cleavage of fetuin alone, whereas 10F4 inhibited cleavage of both substrates. All four MAbs reduced the in vitro spread of viruses carrying either the wild-type N9 or N9 with antiviral-resistant mutations but to different degrees. These MAbs have different in vivo levels of effectiveness: 10F4 was the most effective in protecting mice against challenge with A(H7N9) virus, 2F6 was less effective, and 11B2 failed to protect BALB/c mice at the doses tested. Our study confirms that NA-specific antibodies can protect against A(H7N9) infection and suggests that in vitro properties can be used to rank antibodies with therapeutic potential. IMPORTANCEThe novel A(H7N9) viruses that emerged in China in 2013 continue to infect humans, with a high fatality rate. The most recent outbreak resulted in a larger number of human cases than previous epidemic waves. Due to the absence of a licensed vaccine and the emergence of drug-resistant viruses, there is a need to develop alternative approaches to prevent or treat A(H7N9) infection. We have made a panel of mouse monoclonal antibodies (MAbs) specific for neuraminidase (NA) of A(H7N9) viruses; some of these MAbs are effective in inhibiting viruses that are resistant to antivirals used to treat A(H7N9) patients. Binding avidity, inhibition of NA activity, and plaque formation correlated with the effectiveness of these MAbs to protect mice against lethal A(H7N9) virus challenge. This study identifies in vitro measures that can be used to predict the in vivo efficacy of NA-specific antibodies, providing a way to select MAbs for further therapeutic development.

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“…Viruses and sequence analysis. The isolation of WT and NA variants (R292K, E119V, and I222K) from the A/Taiwan/1/2013 (H7N9) isolate and their full characterization, including the NA sequence, were previously reported (22).…”

mentioning

confidence: 99%

“…In a single isolate, we identified four virus variants differing by a single amino acid in the NA compared to the wild-type (WT) virus (22). When tested in the fluorescent NI assay with oseltamivir carboxylate, these viruses exhibited a wide range of IC 50 s. The R292K NA variant showed a Ͼ10,000-fold increase in the IC 50 and was defined as a highly reduced (Ͼ100-fold) inhibited virus.…”

mentioning

confidence: 99%

Neuraminidase Mutations Conferring Resistance to Oseltamivir in Influenza A(H7N9) Viruses

Marjuki

Mishin

Chesnokov

et al. 2015

J Virol

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Human infections by avian influenza A(H7N9) virus entail substantial morbidity and mortality. Treatment of infected patients with the neuraminidase (NA) inhibitor oseltamivir was associated with emergence of viruses carrying NA substitutions. In the NA inhibition (NI) assay, R292K conferred highly reduced inhibition by oseltamivir, while E119V and I222K each caused reduced inhibition. To facilitate establishment of laboratory correlates of clinically relevant resistance, experiments were conducted in ferrets infected with virus carrying wild-type or variant NA genes recovered from the A/Taiwan/ 1/2013 isolate. Oseltamivir treatment (5 or 25 mg/kg of body weight/dose) was given 4 h postinfection, followed by twicedaily treatment for 5 days. Treatment of ferrets infected with wild-type virus resulted in a modest dose-dependent reduction (0.7 to 1.5 log 10 50% tissue culture infectious dose [TCID 50 ]) in nasal wash viral titers and inflammation response. Conversely, treatment failed to significantly inhibit the replication of R292K or E119V virus. A small reduction of viral titers was detected on day 5 in ferrets infected with the I222K virus. The propensity for oseltamivir resistance emergence was assessed in oseltamivir-treated animals infected with wild-type virus; emergence of R292K virus was detected in 3 of 6 ferrets within 5 to 7 days postinfection. Collectively, we demonstrate that R292K, E119V, and I222K reduced the inhibitory activity of oseltamivir, not only in the NI assay, but also in infected ferrets, judged particularly by viral loads in nasal washes, and may signal the need for alternative therapeutics. Thus, these clinical outcomes measured in the ferret model may correlate with clinically relevant oseltamivir resistance in humans. IMPORTANCEThis report provides more evidence for using the ferret model to assess the susceptibility of influenza A(H7N9) viruses to oseltamivir, the most prescribed anti-influenza virus drug. The information gained can be used to assist in the establishment of laboratory correlates of human disease and drug therapy. The rapid emergence of viruses with R292K in treated ferrets correlates well with the multiple reports on this NA variant in treated human patients. Our findings highlight the importance of the discovery and characterization of new antiviral drugs with different mechanisms of action and the use of combination treatment strategies against emerging viruses with pandemic potential, such as avian H7N9 virus, particularly against those carrying drug resistance markers.

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Characterization of Drug-Resistant Influenza A(H7N9) Variants Isolated From an Oseltamivir-Treated Patient in Taiwan

Abstract: Our data highlight challenges in assessment of the replicative fitness of H7N9 NA variants that emerged in NAI-treated patients.

Cited by 73 publications

References 43 publications

The therapeutic effects of sodium cromoglycate against influenza A virus H5N1 in mice

The therapeutic effects of sodium cromoglycate against influenza A virus H5N1 in mice

Comparison of the Efficacy of N9 Neuraminidase-Specific Monoclonal Antibodies against Influenza A(H7N9) Virus Infection

Neuraminidase Mutations Conferring Resistance to Oseltamivir in Influenza A(H7N9) Viruses

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