Mutations to PB2 and NP proteins of an avian influenza virus combine to confer efficient growth in primary human respiratory cells

Shamika Danzy; Lydia R Studdard; Balaji Manicassamy; Alicia Solorzano; Nicolle Marshall; Adolfo García-Sastre; John Steel; Anice C Lowen

doi:10.1128/JVI.01093-14

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Mutations to PB2 and NP proteins of an avian influenza virus combine to confer efficient growth in primary human respiratory cells

Journal article

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Mutations to PB2 and NP proteins of an avian influenza virus combine to confer efficient growth in primary human respiratory cells

Shamika Danzy, Lydia R Studdard, Balaji Manicassamy, Alicia Solorzano, Nicolle Marshall, Adolfo García-Sastre, John Steel and Anice C Lowen

Journal of virology, Vol.88(22), pp.13436-13446

11/2014

DOI: 10.1128/JVI.01093-14

PMCID: PMC4249088

PMID: 25210184

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Abstract

Influenza pandemics occur when influenza A viruses (IAV) adapted to other host species enter humans and spread through the population. Pandemics are relatively rare due to host restriction of IAV: strains adapted to nonhuman species do not readily infect, replicate in, or transmit among humans. IAV can overcome host restriction through reassortment or adaptive evolution, and these are mechanisms by which pandemic strains arise in nature. To identify mutations that facilitate growth of avian IAV in humans, we have adapted influenza A/duck/Alberta/35/1976 (H1N1) (dk/AB/76) virus to a high-growth phenotype in differentiated human tracheo-bronchial epithelial (HTBE) cells. Following 10 serial passages of three independent lineages, the bulk populations showed similar growth in HTBE cells to that of a human seasonal virus. The coding changes present in six clonal isolates were determined. The majority of changes were located in the polymerase complex and nucleoprotein (NP), and all isolates carried mutations in the PB2 627 domain and regions of NP thought to interact with PB2. Using reverse genetics, the impact on growth and polymerase activity of individual and paired mutations in PB2 and NP was evaluated. The results indicate that coupling of the mammalian-adaptive mutation PB2 E627K or Q591K to selected mutations in NP further augments the growth of the corresponding viruses. In addition, minimal combinations of three (PB2 Q236H, E627K, and NP N309K) or two (PB2 Q591K and NP S50G) mutations were sufficient to recapitulate the efficient growth in HTBE cells of dk/AB/76 viruses isolated after 10 passages in this substrate. Influenza A viruses adapted to birds do not typically grow well in humans. However, as has been seen recently with H5N1 and H7N9 subtype viruses, productive and virulent infection of humans with avian influenza viruses can occur. The ability of avian influenza viruses to adapt to new host species is a consequence of their high mutation rate that supports their zoonotic potential. Understanding of the adaptation of avian viruses to mammals strengthens public health efforts aimed at controlling influenza. In particular, it is critical to know how readily and through mutation to which functional components avian influenza viruses gain the ability to grow efficiently in humans. Our data show that as few as three mutations, in the PB2 and NP proteins, support robust growth of a low-pathogenic, H1N1 duck isolate in primary human respiratory cells.

Mutation

Adaptation, Biological

Animals

Cell Line

DNA Mutational Analysis

Ducks

Epithelial Cells - virology

Humans

Influenza A Virus, H1N1 Subtype - genetics

Influenza A Virus, H1N1 Subtype - growth & development

Influenza A Virus, H1N1 Subtype - physiology

Influenza in Birds - virology

Mutant Proteins - genetics

Mutant Proteins - metabolism

Recombination, Genetic

Reverse Genetics

RNA Replicase - genetics

RNA Replicase - metabolism

RNA-Binding Proteins - genetics

RNA-Binding Proteins - metabolism

Serial Passage

Viral Core Proteins - genetics

Viral Core Proteins - metabolism

Viral Proteins - genetics

Viral Proteins - metabolism

Details

Title: Subtitle: Mutations to PB2 and NP proteins of an avian influenza virus combine to confer efficient growth in primary human respiratory cells
Creators: Shamika Danzy - Emory University School of Medicine
Lydia R Studdard - Emory University School of Medicine
Balaji Manicassamy - University of Iowa, Microbiology and Immunology
Alicia Solorzano - Rutgers, The State University of New Jersey
Nicolle Marshall - Emory University School of Medicine
Adolfo García-Sastre - Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USA Department of Medicine, Division of Infectious Diseases, Icahn School of Medicine at Mount Sinai, New York, New York, USA Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA
John Steel - Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA
Anice C Lowen - Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia, USA anice.lowen@emory.edu
Resource Type: Journal article
Publication Details: Journal of virology, Vol.88(22), pp.13436-13446
DOI: 10.1128/JVI.01093-14
PMID: 25210184
PMCID: PMC4249088
ISSN: 0022-538X
eISSN: 1098-5514
Grant note: HHSN272201400008C / NIAID NIH HHS HHSN266200700006C / PHS HHS AI095320 / NIAID NIH HHS HHSN272201400008C / PHS HHS HHSN272201400004C / NIAID NIH HHS HHSN2662007000010C / PHS HHS K99 AI095320 / NIAID NIH HHS R00 AI095320 / NIAID NIH HHS HHSN272201400004C / PHS HHS UL1 TR000430 / NCATS NIH HHS HHSN266200700010C / NIAID NIH HHS HHSN266200700006C / NIAID NIH HHS
Language: English
Date published: 11/2014
Academic Unit: Microbiology and Immunology
Record Identifier: 9984208754102771

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