Mutations at positions 186 and 194 in the HA gene of the 2009 H1N1 pandemic influenza virus improve replication in cell culture and eggs

Suphaphiphat, Pirada; Franti, Michael; Hekele, Armin; Lilja, Anders; Spencer, Terika; Settembre, Ethan C.; Palmer, Gene A.; Crotta, Stefania; Tuccino, Annunziata Barbara; Keiner, Bjoern; Trusheim, Heidi; Balabanis, Kara; Sackal, Melissa; Rothfeder, Mithra; Mandl, Christian W.; Dormitzer, Philip R.; Mason, Peter W.

doi:10.1186/1743-422x-7-157

Cited by 21 publications

(19 citation statements)

References 24 publications

(24 reference statements)

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“…Indeed, a previous study has found serine in this position to be a human H1N1 characteristic, whereas proline in this position characterizes swine strains (20). Moreover, this position has been recently reported to affect growth in cell culture and eggs (21). Thus, the appearance of serine in pH1N1 may be one of the characteristics leading it to become a more "human-like" virus.…”

Section: Resultsmentioning

confidence: 99%

Putative amino acid determinants of the emergence of the 2009 influenza A (H1N1) virus in the human population

Meroz

Yoon

Ducatez

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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The emergence of the unique H1N1 influenza A virus in 2009 resulted in a pandemic that has spread to over 200 countries. The constellation of molecular factors leading to the emergence of this strain is still unclear. Using a computational approach, we identified molecular determinants that may discriminate the hemagglutinin protein of the 2009 human pandemic H1N1 (pH1N1) strain from that of other H1N1 strains. As expected, positions discriminating the pH1N1 from seasonal human strains were located in or near known H1N1 antigenic sites, thus camouflaging the pH1N1 strain from immune recognition. For example, the alteration S145K (an antigenic position) was found as a characteristic of the pH1N1 strain. We also detected positions in the hemagglutinin protein differentiating classical swine viruses from pH1N1. These positions were mostly located in and around the receptor-binding pocket, possibly influencing binding affinity to the human cell. Such alterations may be liable in part for the virus's efficient infection and adaptation to humans. For instance, 133 A and 149 were identified as discriminative positions. Significantly, we showed that the substitutions R133 A K and R149K, predicted to be pH1N1 characteristics, each altered virus binding to erythrocytes and conferred virulence to A/swine/NC/18161/02 in mice, reinforcing the computational findings. Our findings provide a structural explanation for the deficient immunity of humans to the pH1N1 strain. Moreover, our analysis points to unique molecular factors that may have facilitated the emergence of this swine variant in humans, in contrast to other swine variants that failed.A pandemic H1N1 (pH1N1) human influenza virus was identified in April 2009 (1) and has since spread to over 200 countries and caused over 18,000 deaths (2). Evolutionary analysis of the pH1N1 strain indicates that its HA belongs to the classical swine lineage. HAs in this lineage are more similar to those of historical human strains such as the 1918 H1N1 strain than to those of circulating H1N1 strains from recent years (3). Prior to the emergence of the pH1N1 strain, only sporadic cases of human infection by swine influenza viruses had been reported (4-8). The molecular basis enabling this recent strain to efficiently infect and be transmitted between humans remains obscure (9-12).Herein, we used a computational approach to uncover signature residues in the receptor-binding domain (RBD) of the pH1N1 HA protein. Using a machine-learning algorithm of alternating decision trees (ADTs) (13), we predicted positions that differentiated between HA protein sequences of the pH1N1 strain and HA sequences of other H1N1 strains. The algorithm combines moderately successful rules to produce highly accurate predictions (13). Here the rules represent correlations between the presence of specific amino acid type(s) in selected positions and the sequence annotation, e.g., pandemic versus seasonal strain. This is done by iteratively reweighting training examples (i.e., HA sequences), thus, concentratin...

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Section: Resultsmentioning

confidence: 99%

Putative amino acid determinants of the emergence of the 2009 influenza A (H1N1) virus in the human population

Meroz

Yoon

Ducatez

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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“…1A), although HA was indeed present and recognized by an anti-pH1N1 MAb (data not shown). The original isolate of the pH1N1 virus does not replicate well in MDCK cells but can be made to propagate well in vitro if critical mutations are incorporated into the receptor-binding pocket of HA (13,27,28). An example is the pH1N1/E3 virus generated in our laboratory by selection of a high-growth clone of pH1N1 in 10-day-old embryonated chicken eggs (13).…”

Section: Resultsmentioning

confidence: 99%

Protection against Lethal Influenza with a Viral Mimic

Baker

Guo

Albrecht

et al. 2013

J Virol

100

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Despite countermeasures against influenza virus that prevent (vaccines) and treat (antivirals) infection, this upper respiratory tract human pathogen remains a global health burden, causing both seasonal epidemics and occasional pandemics. More potent and safe new vaccine technologies would contribute significantly to the battle against influenza and other respiratory infections. Using plasmid-based reverse genetics techniques, we have developed a single-cycle infectious influenza virus (sciIV) with immunoprotective potential. In our sciIV approach, the fourth viral segment, which codes for the receptor-binding and fusion protein hemagglutinin (HA), has been removed. Thus, upon infection of normal cells, although no infectious progeny are produced, the expression of other viral proteins occurs and is immunogenic. Consequently, sciIV is protective against influenza homologous and heterologous viral challenges in a mouse model. Vaccination with sciIV protects in a dose-and replication-dependent manner, which is attributed to both humoral responses and T cells. Safety, immunogenicity, and protection conferred by sciIV vaccination were also demonstrated in ferrets, where this immunization additionally blocked direct and aerosol transmission events. All together, our studies suggest that sciIV may have potential as a broadly protective vaccine against influenza virus.

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“…Sequencing of our CA/E3/09 virus revealed three mutations in HA (K136N, S200P, and D239G). Due to differences in the numbering system chosen, the S200P mutation corresponds to the S186P mutation in the plasmid F8 genetic mutant CA/09 virus created by Suphaphiphat et al, which has been shown to increase the growth of the virus in either eggs or MDCK cells (36). The single mutation of S to P at position 186 does not change the antigenicity of the virus (36).…”

Section: Discussionmentioning

confidence: 99%

“…Due to differences in the numbering system chosen, the S200P mutation corresponds to the S186P mutation in the plasmid F8 genetic mutant CA/09 virus created by Suphaphiphat et al, which has been shown to increase the growth of the virus in either eggs or MDCK cells (36). The single mutation of S to P at position 186 does not change the antigenicity of the virus (36). In addition, the K136N mutation in our stock virus is in the same position as the K119N mutation found in an MDCK cell-adapted CA/ 07/09 virus (11).…”

Section: Discussionmentioning

confidence: 99%

T Cell-Mediated Protection against Lethal 2009 Pandemic H1N1 Influenza Virus Infection in a Mouse Model

Guo

Santiago

Lambert

et al. 2011

J Virol

121

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Mutations at positions 186 and 194 in the HA gene of the 2009 H1N1 pandemic influenza virus improve replication in cell culture and eggs

Cited by 21 publications

References 24 publications

Putative amino acid determinants of the emergence of the 2009 influenza A (H1N1) virus in the human population

Putative amino acid determinants of the emergence of the 2009 influenza A (H1N1) virus in the human population

Protection against Lethal Influenza with a Viral Mimic

T Cell-Mediated Protection against Lethal 2009 Pandemic H1N1 Influenza Virus Infection in a Mouse Model

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