Structural and evolutionary characteristics of HA, NA, NS and M genes of clinical influenza A/H3N2 viruses passaged in human and canine cells

Жирнов, О. П.; Vorobjeva, I V; Saphonova, O A; Poyarkov, Stanislav; Овчаренко, А. В.; Anhlan, Darisuren; Malyshev, N A

doi:10.1016/j.jcv.2009.05.030

Cited by 41 publications

(40 citation statements)

References 42 publications

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“…Substitutions at two other residues, 136 and 151, in the NA of A(H1N1) have been shown to affect drug NA inhibitor susceptibility and were linked to influenza virus propagation in MDCK cells (31,32). Moreover, previous studies have shown that the MDCK cell-grown A(H3N2) virus contained substitutions at positions 148 and 151, whereas its counterpart propagated in CaCo-2 cells did not have such changes (10). Despite the growing evidence regarding selection of NA variants in MDCK cells, the cell line continues to be the most common substrate for the propagation of human influenza viruses.…”

Section: Discussionmentioning

confidence: 99%

“…Many cell lines have been used for the propagation of influenza viruses, including baby hamster kidney (BHK-21) cells, rhesus monkey kidney epithelial (LLC-MK2) cells, and African green monkey kidney (Vero) cells (5)(6)(7)(8). The use of human colon intestinal epithelial (CaCo-2) cells for isolation and propagation of seasonal A(H3N2) viruses has also been advocated in recent years (9,10). Madin-Darby canine kidney (MDCK) cells, however, remain the most commonly used cell line for the propagation of influenza viruses due to their superior overall sensitivity, high virus yield, and ease of culture and maintenance (11)(12)(13)(14).…”

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confidence: 99%

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Cell Culture-Selected Substitutions in Influenza A(H3N2) Neuraminidase Affect Drug Susceptibility Assessment

Tamura

Nguyen

Sleeman

et al. 2013

Antimicrob Agents Chemother

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Assessment of drug susceptibility has become an integral part of influenza virus surveillance. In this study, we describe the drug resistance profile of influenza A(H3N2) virus, A/Mississippi/05/2011, collected from a patient treated with oseltamivir and detected via surveillance. An MDCK cell-grown isolate of this virus exhibited highly reduced inhibition by the neuraminidase (NA) inhibitors (NAIs) oseltamivir (8,005-fold), zanamivir (813-fold), peramivir (116-fold), and laninamivir (257-fold) in the NA inhibition assay. Sequence analysis of its NA gene revealed a known oseltamivir-resistance marker, the glutamic acid-to-valine substitution at position 119 (E119V), and an additional change, threonine to isoleucine at position 148 (T148I). Unlike E119V, T148I was not detected in the clinical sample but acquired during viral propagation in MDCK cells. Using recombinant proteins, T148I by itself was shown to cause only a 6-fold increase in the zanamivir 50% inhibitory concentration (IC 50 ) and had no effect on inhibition by other drugs. The T148I substitution reduced NA activity by 50%, most likely by affecting the positioning of the 150 loop at the NA catalytic site. Using pyrosequencing, changes at T148 were detected in 35 (23%) of 150 MDCK cell-grown A(H3N2) viruses tested, which was lower than the frequency of changes at D151 (85%), an NA residue previously implicated in cell selection. We demonstrate that culturing of the A(H3N2) viruses (n ‫؍‬ 11) at a low multiplicity of infection delayed the emergence of the NA variants with changes at position 148 and/or 151, especially when conducted in MDCK-SIAT1 cells. Our findings highlight the current challenges in monitoring susceptibility of influenza A(H3N2) viruses to the NAI class of antiviral drugs.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Cell Culture-Selected Substitutions in Influenza A(H3N2) Neuraminidase Affect Drug Susceptibility Assessment

Tamura

Nguyen

Sleeman

et al. 2013

Antimicrob Agents Chemother

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“…It is also possible that the M gene of the R lineage was generated by the accumulation of mutations in the M gene of the S lineage during passages in cell lines. Mutations can occur during isolation or passage in cell lines (7,25); however, when we passaged isolates in amantadine after plaque purification, the M gene in the S lineage acquired a point mutation that led to amantadine resistance. The results showed that the de novo accumulation of mutations in the S lineage did not generate the M gene in the R lineage.…”

Section: Discussionmentioning

confidence: 99%

Occurrence of Mixed Populations of Influenza A Viruses That Can Be Maintained through Transmission in a Single Host and Potential for Reassortment

et al. 2010

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Reassortment, which is the rearrangement of viral gene segments in a host cell infected with two different viruses, is an important mechanism for the evolution of influenza viruses. Mixed infections with multiple virus types could lead to reassortment. To better understand the occurrence of quasispecies in a single host, we investigated mixed infections in individual isolates of seasonal influenza A viruses using amantadine sensitivity as a marker. We cultured viruses with amantadine and performed sequencing, restriction fragment length polymorphism analysis, cloning, and quantitative PCR to detect mixed populations. Culturing with amantadine showed evidence of a high number of mixed populations, while the other assays could hardly detect mixed populations. The existence of quasispecies in each isolate was common. However, the proportion of these, which can be less than 1%, is too low to be detected by conventional methods. Such mixed populations in which reassortment occurs may have a significant role in the evolution of viruses.

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“…There has been no O- linked glycosylation (e.g. attachment of a sugar molecule to the hydroxyl group of Ser or Thr on the polypeptide chain) reported for IAV8. The stepwise increase in the number of N- linked glycosylation sites on the H3 globular head has been shown to reduce virulence of H3N2 strains91011.…”

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confidence: 99%

Glycosylation changes in the globular head of H3N2 influenza hemagglutinin modulate receptor binding without affecting virus virulence

Alymova

York

Air

et al. 2016

Sci Rep

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Since the emergence of human H3N2 influenza A viruses in the pandemic of 1968, these viruses have become established as strains of moderate severity. A decline in virulence has been accompanied by glycan accumulation on the hemagglutinin globular head, and hemagglutinin receptor binding has changed from recognition of a broad spectrum of glycan receptors to a narrower spectrum. The relationship between increased glycosylation, binding changes, and reduction in H3N2 virulence is not clear. We evaluated the effect of hemagglutinin glycosylation on receptor binding and virulence of engineered H3N2 viruses. We demonstrate that low-binding virus is as virulent as higher binding counterparts, suggesting that H3N2 infection does not require either recognition of a wide variety of, or high avidity binding to, receptors. Among the few glycans recognized with low-binding virus, there were two structures that were bound by the vast majority of H3N2 viruses isolated between 1968 and 2012. We suggest that these two structures support physiologically relevant binding of H3N2 hemagglutinin and that this physiologically relevant binding has not changed since the 1968 pandemic. Therefore binding changes did not contribute to reduced severity of seasonal H3N2 viruses. This work will help direct the search for factors enhancing influenza virulence.

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Structural and evolutionary characteristics of HA, NA, NS and M genes of clinical influenza A/H3N2 viruses passaged in human and canine cells

Cited by 41 publications

References 42 publications

Cell Culture-Selected Substitutions in Influenza A(H3N2) Neuraminidase Affect Drug Susceptibility Assessment

Cell Culture-Selected Substitutions in Influenza A(H3N2) Neuraminidase Affect Drug Susceptibility Assessment

Occurrence of Mixed Populations of Influenza A Viruses That Can Be Maintained through Transmission in a Single Host and Potential for Reassortment

Glycosylation changes in the globular head of H3N2 influenza hemagglutinin modulate receptor binding without affecting virus virulence

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