Cooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples

ABSTRACT The high mutation rates of RNA viruses lead to rapid genetic diversification, which can enable cooperative interactions between variants in a viral population. We previously described two distinct variants of H3N2 influenza virus that cooperate in cell culture. These variants differ by a si...

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Autores principales: Katherine S. Xue, Alexander L. Greninger, Ailyn Pérez-Osorio, Jesse D. Bloom
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Publicado: American Society for Microbiology 2018
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spelling oai:doaj.org-article:b536cca4400a4c6eb20eb5a6375cf1a12021-11-15T15:22:01ZCooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples10.1128/mSphereDirect.00552-172379-5042https://doaj.org/article/b536cca4400a4c6eb20eb5a6375cf1a12018-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphereDirect.00552-17https://doaj.org/toc/2379-5042ABSTRACT The high mutation rates of RNA viruses lead to rapid genetic diversification, which can enable cooperative interactions between variants in a viral population. We previously described two distinct variants of H3N2 influenza virus that cooperate in cell culture. These variants differ by a single mutation, D151G, in the neuraminidase protein. The D151G mutation reaches a stable frequency of about 50% when virus is passaged in cell culture. However, it is unclear whether selection for the cooperative benefits of D151G is a cell culture phenomenon or whether the mutation is also sometimes present at appreciable frequency in virus populations sampled directly from infected humans. Prior work has not detected D151G in unpassaged clinical samples, but those studies have used methods like Sanger sequencing and pyrosequencing, which are relatively insensitive to low-frequency variation. We identified nine samples of human H3N2 influenza virus collected between 2013 and 2015 in which Sanger sequencing had detected a high frequency of the D151G mutation following one to three passages in cell culture. We deep sequenced the unpassaged clinical samples to identify low-frequency viral variants. The frequency of D151G did not exceed the frequency of library preparation and sequencing errors in any of the sequenced samples. We conclude that passage in cell culture is primarily responsible for the frequent observations of D151G in recent H3N2 influenza virus strains. IMPORTANCE Viruses mutate rapidly, and recent studies of RNA viruses have shown that related viral variants can sometimes cooperate to improve each other’s growth. We previously described two variants of H3N2 influenza virus that cooperate in cell culture. The mutation responsible for cooperation is often observed when human samples of influenza virus are grown in the lab before sequencing, but it is unclear whether the mutation also exists in human infections or is exclusively the result of lab passage. We identified nine human isolates of influenza virus that had developed the cooperating mutation after being grown in the lab and performed highly sensitive deep sequencing of the unpassaged clinical samples to determine whether the mutation existed in the original human infections. We found no evidence of the cooperating mutation in the unpassaged samples, suggesting that the cooperation arises primarily under laboratory conditions.Katherine S. XueAlexander L. GreningerAilyn Pérez-OsorioJesse D. BloomAmerican Society for MicrobiologyarticleD151Gcooperationdeep sequencinginfluenza virusneuraminidasequasispeciesMicrobiologyQR1-502ENmSphere, Vol 3, Iss 1 (2018)
institution DOAJ
collection DOAJ
language EN
topic D151G
cooperation
deep sequencing
influenza virus
neuraminidase
quasispecies
Microbiology
QR1-502
spellingShingle D151G
cooperation
deep sequencing
influenza virus
neuraminidase
quasispecies
Microbiology
QR1-502
Katherine S. Xue
Alexander L. Greninger
Ailyn Pérez-Osorio
Jesse D. Bloom
Cooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples
description ABSTRACT The high mutation rates of RNA viruses lead to rapid genetic diversification, which can enable cooperative interactions between variants in a viral population. We previously described two distinct variants of H3N2 influenza virus that cooperate in cell culture. These variants differ by a single mutation, D151G, in the neuraminidase protein. The D151G mutation reaches a stable frequency of about 50% when virus is passaged in cell culture. However, it is unclear whether selection for the cooperative benefits of D151G is a cell culture phenomenon or whether the mutation is also sometimes present at appreciable frequency in virus populations sampled directly from infected humans. Prior work has not detected D151G in unpassaged clinical samples, but those studies have used methods like Sanger sequencing and pyrosequencing, which are relatively insensitive to low-frequency variation. We identified nine samples of human H3N2 influenza virus collected between 2013 and 2015 in which Sanger sequencing had detected a high frequency of the D151G mutation following one to three passages in cell culture. We deep sequenced the unpassaged clinical samples to identify low-frequency viral variants. The frequency of D151G did not exceed the frequency of library preparation and sequencing errors in any of the sequenced samples. We conclude that passage in cell culture is primarily responsible for the frequent observations of D151G in recent H3N2 influenza virus strains. IMPORTANCE Viruses mutate rapidly, and recent studies of RNA viruses have shown that related viral variants can sometimes cooperate to improve each other’s growth. We previously described two variants of H3N2 influenza virus that cooperate in cell culture. The mutation responsible for cooperation is often observed when human samples of influenza virus are grown in the lab before sequencing, but it is unclear whether the mutation also exists in human infections or is exclusively the result of lab passage. We identified nine human isolates of influenza virus that had developed the cooperating mutation after being grown in the lab and performed highly sensitive deep sequencing of the unpassaged clinical samples to determine whether the mutation existed in the original human infections. We found no evidence of the cooperating mutation in the unpassaged samples, suggesting that the cooperation arises primarily under laboratory conditions.
format article
author Katherine S. Xue
Alexander L. Greninger
Ailyn Pérez-Osorio
Jesse D. Bloom
author_facet Katherine S. Xue
Alexander L. Greninger
Ailyn Pérez-Osorio
Jesse D. Bloom
author_sort Katherine S. Xue
title Cooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples
title_short Cooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples
title_full Cooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples
title_fullStr Cooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples
title_full_unstemmed Cooperating H3N2 Influenza Virus Variants Are Not Detectable in Primary Clinical Samples
title_sort cooperating h3n2 influenza virus variants are not detectable in primary clinical samples
publisher American Society for Microbiology
publishDate 2018
url https://doaj.org/article/b536cca4400a4c6eb20eb5a6375cf1a1
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