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. 2012 Jan 5;422(1):105-13.
doi: 10.1016/j.virol.2011.10.006. Epub 2011 Nov 5.

In vitro evolution of H5N1 avian influenza virus toward human-type receptor specificity

Li-Mei Chen  1 , Ola Blixt, James Stevens, Aleksandr S Lipatov, Charles T Davis, Brian E Collins, Nancy J Cox, James C Paulson, Ruben O Donis
Affiliations

In vitro evolution of H5N1 avian influenza virus toward human-type receptor specificity

Li-Mei Chen et al. Virology. .

Erratum in

  • Virology. 2012 Mar 15;424(2):154

Abstract

Acquisition of α2-6 sialoside receptor specificity by α2-3 specific highly-pathogenic avian influenza viruses (H5N1) is thought to be a prerequisite for efficient transmission in humans. By in vitro selection for binding α2-6 sialosides, we identified four variant viruses with amino acid substitutions in the hemagglutinin (S227N, D187G, E190G, and Q196R) that revealed modestly increased α2-6 and minimally decreased α2-3 binding by glycan array analysis. However, a mutant virus combining Q196R with mutations from previous pandemic viruses (Q226L and G228S) revealed predominantly α2-6 binding. Unlike the wild type H5N1, this mutant virus was transmitted by direct contact in the ferret model although not by airborne respiratory droplets. However, a reassortant virus with the mutant hemagglutinin, a human N2 neuraminidase and internal genes from an H5N1 virus was partially transmitted via respiratory droplets. The complex changes required for airborne transmissibility in ferrets suggest that extensive evolution is needed for H5N1 transmissibility in humans.

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Figures

Fig. 1
Fig. 1
Solid phase receptor binding assay of H3N2 and H5N1 variant viruses compared to wildtype. Binding of sialylglycopolymers 3′SLN-PAA (blue line) and 6′SLN-PAA (red lines) by influenza H3N2 and H5N1 viruses performed as described in Materials and methods section. The x axis denotes the concentration of biotinylated sialoglycan (μM) added to the wells, the y axis indicates the resulting absorbance reading. Data shown are representative from three independent binding experiments.
Fig. 2
Fig. 2
Glycan microarray analysis of H5N1 variant viruses and including dual mutations from previous pandemic viruses. Different types of sialoglycans on the array (x-axis) are indicated by colors; the structure of each numbered glycan is provided in Table S1. Vertical bars denote mean binding signal (fluorescence intensity) and T bar extensions indicate the standard error. Glycan microarray analysis of H5N1 variant viruses and including dual mutations from previous pandemic viruses. Different types of sialoglycans on the array (x-axis) are indicated by colors; the structure of each numbered glycan is provided in Table S1. Vertical bars denote mean binding signal (fluorescence intensity) and T bar extensions indicate the standard error.
Fig. 3
Fig. 3
Solid phase receptor binding assay of H5N1 variant with dual mutations close to the receptor binding site. Binding of sialylglycopolymers 3′SLN-PAA (blue line) and 6′SLN-PAA (red lines) by influenza H5N1 variants was described in Materials and methods section. The x and y axis are as in Fig. 1. Data shown are representative from three independent binding experiments.
Fig. 4
Fig. 4
Glycan microarray analysis of clade 2 H5N1 viruses with dual mutations close to the receptor binding site. The color conventions and glycan key for the charts are described in Fig. 2.
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References

    1. Blixt O, Head S, Mondala T, Scanlan C, Huflejt ME, Alvarez R, Bryan MC, Fazio F, Calarese D, Stevens J, Razi N, Stevens DJ, Skehel JJ, van Die I, Burton DR, Wilson IA, Cummings R, Bovin N, Wong CH, Paulson JC. Printed covalent glycan array for ligand profiling of diverse glycan binding proteins. Proc Natl Acad Sci U S A. 2004;101:17033–17038. - PMC - PubMed
    1. Carroll SM, Paulson JC. Differential infection of receptor-modified host cells by receptor-specific influenza viruses. Virus Res. 1985;3:165–179. - PubMed
    1. Chen LM, Rivailler P, Hossain J, Carney P, Balish A, Perry I, Davis CT, Garten R, Shu B, Xu X, Klimov A, Paulson JC, Cox NJ, Swenson S, Stevens J, Vincent A, Gramer M, Donis RO. Receptor specificity of subtype H1 influenza A viruses isolated from swine and humans in the United States. Virology. 2011;412:401–410. - PMC - PubMed
    1. Chutinimitkul S, van Riel D, Munster VJ, van den Brand JM, Rimmelzwaan GF, Kuiken T, Osterhaus AD, Fouchier RA, de Wit E. In vitro assessment of attachment pattern and replication efficiency of H5N1 influenza A viruses with altered receptor specificity. J Virol. 2010;84:11802–11813. - PMC - PubMed
    1. Collins BEBE, Blixt OO, Han SS, Duong BB, Li HH, Nathan JKJK, Bovin NN, Paulson JCJC. High-affinity ligand probes of CD22 overcome the threshold set by cis ligands to allow for binding, endocytosis, and killing of B cells. J Immunol. 2006;177:2994–3003. - PubMed

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