1. Immunology and Inflammation
  2. Microbiology and Infectious Disease

A paucigranulocytic asthma host environment promotes the emergence of virulent influenza viral variants

  1. Katina D Hulme
  2. Anjana C Karawita
  3. Cassandra Pegg
  4. Myrna JM Bunte
  5. Helle Bielefeldt-Ohmann
  6. Conor J Bloxham
  7. Silvie Van den Hoecke
  8. Yin Xiang Setoh
  9. Bram Vrancken
  10. Monique Spronken
  11. Lauren E Steele
  12. Nathalie AJ Verzele
  13. Kyle R Upton
  14. Alexander A Khromykh
  15. Keng Yih Chew
  16. Maria Sukkar
  17. Simon Phipps
  18. Kirsty R Short  Is a corresponding author
  1. School of Chemistry and Molecular Biosciences, The University of Queensland, Australia
  2. Australian Infectious Diseases Research Centre, The University of Queensland, Australia
  3. School of Veterinary Science, The University of Queensland, Australia
  4. School of Biomedical Sciences, The University of Queensland, Australia
  5. VIB-UGent Center for Medical Biotechnology, Belgium
  6. Department of Biomedical Molecular Biology, Ghent University, Belgium
  7. Environmental Health Institute, National Environment Agency, Singapore
  8. KU Leuven, Department of Microbiology and Immunology, Rega Institute, Laboratory of Evolutionary and Computational Virology, Belgium
  9. Department of Viroscience, Erasmus MC, Netherlands
  10. Discipline of Pharmacy, Graduate School of Health, University of Technology Sydney, Australia; Woolcock Institute of Medical Research, Sydney Medical School, University of Sydney, Australia
  11. QIMR Berghofer Medical Research Institute, Australia
https://doi.org/10.7554/eLife.61803
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Cite this article
  1. Katina D Hulme
  2. Anjana C Karawita
  3. Cassandra Pegg
  4. Myrna JM Bunte
  5. Helle Bielefeldt-Ohmann
  6. Conor J Bloxham
  7. Silvie Van den Hoecke
  8. Yin Xiang Setoh
  9. Bram Vrancken
  10. Monique Spronken
  11. Lauren E Steele
  12. Nathalie AJ Verzele
  13. Kyle R Upton
  14. Alexander A Khromykh
  15. Keng Yih Chew
  16. Maria Sukkar
  17. Simon Phipps
  18. Kirsty R Short
(2021)
A paucigranulocytic asthma host environment promotes the emergence of virulent influenza viral variants
eLife 10:e61803.
https://doi.org/10.7554/eLife.61803
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10 figures, 1 table and 4 additional files

Figures

Figure 1
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Pneumonia virus of mice (PVM)-infection (at 7 days of age and reinfection at 35 days of age) of RAGE-deficient mice induces the cardinal features of asthma.

(A) (Left) Percentage of airway epithelial cells (AECs) producing mucus was quantified by immunohistochemistry at 7 days post-reinfection (i.e. 42 days of age) from uninfected RAGE-deficient mice …

Figure 2
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RAGE-deficient asthmatic mice experience more severe influenza than non-asthmatic mice.

RAGE-deficient non-asthmatic and asthmatic mice were infected with 100 (PFU) of H1N1/Auck/09. Mock control groups received PBS only. (A) Percentage weight loss of influenza A virus (IAV) or mock …

Figure 3 with 1 supplement
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Asthmatic hosts produced more diverse viral variants in PB1.

Influenza viral RNA isolated from the lung tissue of asthmatic (n = 27) and non-asthmatic (n = 25) mice was analysed for viral variants at 4 and 6 days post-infection (d.p.i). (A–H) Measurement of …

Figure 3—figure supplement 1
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Read depth of the eight Influenza genome segments from Ager-/- mice.

(A–H) Influenza viral RNA was isolated from the lung tissue of either asthmatic (n = 27) and non-asthmatic (n = 25) mice and analysed for viral variants. Each data point represents a sample. …

Figure 4
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Asthmatic hosts produced more single nucleotide variants (SNVs) in the PB1 genome segment.

Influenza viral RNA isolated from the lung tissue of asthmatic (n = 27) and non-asthmatic (n = 25) mice was analysed for viral variants. Each data point represents the number of viral variants …

Figure 5
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Increased replication of influenza A virus (IAV) was observed in samples derived from asthmatic hosts in vitro.

IAV isolated from asthmatic hosts replicates more quickly than non-asthmatic hosts in (A) Madin–Darby canine kidney (MDCK) cells, (B) Immortalized Mouse Mammary Epithelial Cells (iMMEC), and (C) …

Figure 6
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Crystal structures of the mutations within PB1.

(A) Mutations C693Y and K691R. The molecule is displayed in surface view with cartoon representation of the protein backbone (PB1 magenta and PB2 sky blue). The α-helices are labelled and PB1 C693Y …

Figure 7
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The PB1 C694Y and P651Q mutations showed increased polymerase activity compared to the wild type (WT).

Influenza RNA polymerase-driven activity is increased compared to WT in HEK 293 T cells transfected with either the C693Y or P651Q mutation in PB1. Data are pooled from four independent experiments …

Figure 8
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Mice infected with asthmatic host-derived virus experience more severe influenza than mice infected with non-asthmatic host-derived virus.

C57BL/6 mice were infected with 2000 PFU of either asthmatic or non-asthmatic host-derived influenza A virus (IAV). (A) Percentage weight loss of mice following IAV infection. Weights are displayed …

Figure 9
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Asthmatic host-derived virus shows consolidation of PB1 viral variants population across parent and daughter.

C57BL/6 mice were infected with 6 days post-infection (d.p.i.) murine lung homogenate from either non-asthmatic or asthmatic hosts. Multiple dimension scaling (MDS) analysis of the PB1 gene in (A) …

Author response image 1
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Asthmatic day 6 samples show a positive relationship between weight loss and measures of viral diversity in the PB1.

Influenza viral RNA isolated from the lung tissue of asthmatic (n=27) and non-asthmatic (n=25) mice was analysed for viral variants. Weight-loss as a function of (A) Shannon entropy of PB1 genome …

Tables

Table 1
Frequency of exclusive non-synonymous viral mutations detected in PB1 at 4 and 6 days post-influenza A virus (IAV) inoculation and their frequency in clinical samples.
Non-asthmaticAsthmaticClinical samples (n = 51 per group)
Day 4 (n* = 13)Day 6 (n = 12)Day 4 (n = 13)Day 6 (n = 14)Non-asthmaticAsthmaticp-Value‡
M1I50%†0%0%
M1L17%0%0%
D2H14%0%0%
N77I14%0%0%
P74A14%0%0%
E75D29%0%0%
E75V14%0%0%
D76A14%0%0%
D76H14%0%0%
N77V14%0%0%
E78G14%0%0%
S80H14%0%0%
I205M14%0%0%
I248V14%0%0%
M616I14%0%0%
R621P14%0%0%
C625S14%0%0%
A652P14%0%0%
S654N14%4%18%0.0514
Y657H14%0%0%
N671H17%R670S14%0%0%
 R672P15%0%0%
S673A17%0%0%
I682F15%0%0%
D685E15%0%0%
D685H14%0%0%
M688I14%0%0%
Y689D21%0%0%
Y689R14%0%0%
Q690S15%0%0%
K691R15%4%16%0.092
C692S14%0%0%
C693N14%0%0%
N694R14%0%0%
L695I15%0%0%
R727T14%0%0%

  1. *n = Total number of mouse samples.

    †Percentage of mice with identified mutation.

  2. ‡Statistical comparisons were made using a two-sided fisher’s exact test.

Additional files

Supplementary file 1

Forward and reverse primers used for qPCR.

https://cdn.elifesciences.org/articles/61803/elife-61803-supp1-v1.docx
Supplementary file 2

Non-synonymous viral mutations detected in PB1 at 4 and 6 days post-influenza A virus (IAV) inoculation.

https://cdn.elifesciences.org/articles/61803/elife-61803-supp2-v1.docx
Supplementary file 3

Workflow used to reconstruct haplotypes.

https://cdn.elifesciences.org/articles/61803/elife-61803-supp3-v1.png
Transparent reporting form
https://cdn.elifesciences.org/articles/61803/elife-61803-transrepform-v1.docx

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