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Volume 32, Number 5—May 2026

Dispatch

Replication Efficiency of Contemporary Highly Pathogenic Avian Influenza A(H5N1) Virus Isolates in Human Nasal Epithelium Model

Meaghan Flagg1, Christopher J. Winski1, Bridget G. Brackney, Tessa R. Lutterman, Johan A. Ortiz-Morales2, Brandi N. Williamson, and Emmie de WitComments to Author 
Author affiliation: Author affiliation: National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana, USA

Main Article

Figure 2

Induction of interferon-stimulated genes in human nasal epithelium infected with seasonal influenza A and highly pathogenic avian influenza (HPAI) A(H5N1) viruses in study of replication efficiency of contemporary HPAI H5N1 virus isolates in human nasal epithelium model. We inoculated Mattek EpiNasal cultures (https://www.mattek.com) with a multiplicity of infection of 0.1 50% tissue culture infectious dose per cell and extracted RNA from cells at 0, 8, 24, 48, 72, and 96 hours postinoculation. We ran quantitative reverse transcription PCR using primers (Integrated DNA Technologies, https://www.idtdna.com) to detect interferon-stimulated gene 15 (A), interferon-induced transmembrane protein 3 (B), and myxovirus resistance 1 (C) for historical and clade 2.3.4.4b highly pathogenic avian HPAI H5N1 genotype B3.13, B3.6, and D1.1 virus strains. Lines indicate median; shading indicates 95% CI. We normalized data to internal controls (ACTB and GAPDH) and calculated fold change relative to timepoint-matched mock-infected controls. Fold change is reported for 3 biologic replicates. We performed statistical analysis using 2-way analysis of variance with Dunnett posttest. p values are shown by asterisks in colors matching isolates: *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Figure 2. Induction of interferon-stimulated genes in human nasal epithelium infected with seasonal influenza A and highly pathogenic avian influenza (HPAI) A(H5N1) viruses in study of replication efficiency of contemporary HPAI H5N1 virus isolates in human nasal epithelium model. We inoculated Mattek EpiNasal cultures (https://www.mattek.com) with a multiplicity of infection of 0.1 50% tissue culture infectious dose per cell and extracted RNA from cells at 0, 8, 24, 48, 72, and 96 hours postinoculation. We ran quantitative reverse transcription PCR using primers (Integrated DNA Technologies, https://www.idtdna.com) to detect interferon-stimulated gene 15 (A), interferon-induced transmembrane protein 3 (B), and myxovirus resistance 1 (C) for historical and clade 2.3.4.4b highly pathogenic avian HPAI H5N1 genotype B3.13, B3.6, and D1.1 virus strains. Lines indicate median; shading indicates 95% CI. We normalized data to internal controls (ACTB and GAPDH) and calculated fold change relative to timepoint-matched mock-infected controls. Fold change is reported for 3 biologic replicates. We performed statistical analysis using 2-way analysis of variance with Dunnett posttest. p values are shown by asterisks in colors matching isolates: *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

Main Article

1These authors contributed equally to this article.

2Current affiliation: LSU Health, Shreveport, Louisiana, USA.

Page created: April 14, 2026
Page updated: May 01, 2026
Page reviewed: May 01, 2026
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