Expression of the nonstructural protein NS1 of equine influenza A virus: detection of anti-NS1 antibody in post infection equine sera.
Abstract: The nucleotide sequence of the nonstructural protein NS1 of the influenza virus A/equine 2/Suffolk/89 was determined and found to be 97% identical to that of A/equine 2/Miami/63. A similar level of identity was shown for the deduced NS1 amino acid sequence. The NS1 gene was expressed, in its entirety and in part, as fusion proteins with glutathione S-transferase using the pGEX-3X expression vector. Antibodies to NS1 protein were detected in serum samples from ponies experimentally infected with influenza virus, but not in animals vaccinated with whole inactivated virus or in unprimed control animals. The antigenic determinant(s) of NS1 protein appear to be located in the C-terminal half of the protein. The implications of these findings are discussed with reference to the use of NS1 protein as a differential diagnostic marker for influenza virus infection in the presence of high levels of circulating antibody to influenza haemagglutinin generated by recent vaccination.
Publication Date: 1997-05-01 PubMed ID: 9186949DOI: 10.1016/s0166-0934(97)02189-7Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research focuses on the nonstructural protein NS1 of equine influenza A virus, its genetic similarity to another strain, and its role as a potential diagnostic marker in infected horses, particularly those recently vaccinated against equine influenza.
Study on the NS1 Protein of Equine Influenza A Virus
- The research revolves around exploring the nucleotide sequence of a nonstructural protein NS1 of the influenza virus strain A/equine 2/Suffolk/89. The researchers have discovered that this sequence is 97% identical to the NS1 of another influenza strain (A/equine 2/Miami/63).
- The NS1 gene was expressed either in its entirety or partially as a fusion protein with glutathione S-transferase, a powerful antioxidant, using the pGEX-3X expression vector. A vector is a DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell.
Detection of Antibodies to NS1
- This study also found that antibodies to the NS1 protein were only present in serum samples collected from ponies that were purposefully infected with the influenza virus.
- The research notes that these antibodies were not detected in ponies that had been vaccinated with the whole inactivated virus or in control animals that had not been previously exposed to the virus.
Antigenic Determinants of NS1 Protein
- The research identifies that the antigenic determinant, or the part of the antigen that is recognized by the immune system, of the NS1 protein seems to be located in the C-terminal half of the protein.
Use of NS1 Protein for Diagnostics
- The researchers propose that these findings could have implications for the use of the NS1 protein as a differential diagnostic marker for influenza virus infection. This could be particularly useful in horses that show high levels of circulating antibody to influenza haemagglutinin (a glycoprotein found on the surface of influenza viruses that helps the virus to enter the host cell) due to recent vaccination.
Cite This Article
APA
Birch-Machin I, Rowan A, Pick J, Mumford J, Binns M.
(1997).
Expression of the nonstructural protein NS1 of equine influenza A virus: detection of anti-NS1 antibody in post infection equine sera.
J Virol Methods, 65(2), 255-263.
https://doi.org/10.1016/s0166-0934(97)02189-7 Publication
Researcher Affiliations
- Centre for Preventive Medicine, Animal Health Trust, Kennett, Suffolk, UK.
MeSH Terms
- Amino Acid Sequence
- Animals
- Antibodies, Viral / blood
- Base Sequence
- Chick Embryo
- Genes, Viral
- Glutathione Transferase / genetics
- Glutathione Transferase / immunology
- Horses
- Immunoblotting
- Influenza A virus / chemistry
- Influenza A virus / genetics
- Influenza A virus / immunology
- Molecular Sequence Data
- Orthomyxoviridae Infections / blood
- Orthomyxoviridae Infections / genetics
- Orthomyxoviridae Infections / immunology
- Recombinant Fusion Proteins / biosynthesis
- Recombinant Fusion Proteins / immunology
- Sequence Analysis, DNA
- Viral Nonstructural Proteins / biosynthesis
- Viral Nonstructural Proteins / genetics
- Viral Nonstructural Proteins / immunology
- Viral Structural Proteins / genetics
Citations
This article has been cited 10 times.- Nogales A, DeDiego ML, Martínez-Sobrido L. Live attenuated influenza A virus vaccines with modified NS1 proteins for veterinary use. Front Cell Infect Microbiol 2022;12:954811.
- Sun Z, Wang Q, Li G, Li J, Chen S, Qin T, Ma H, Peng D, Liu X. Development of an Inactivated H7N9 Subtype Avian Influenza Serological DIVA Vaccine Using the Chimeric HA Epitope Approach. Microbiol Spectr 2021 Oct 31;9(2):e0068721.
- Jiang W, Wang Q, Chen S, Gao S, Song L, Liu P, Huang W. Influenza A virus NS1 induces G0/G1 cell cycle arrest by inhibiting the expression and activity of RhoA protein. J Virol 2013 Mar;87(6):3039-52.
- Dundon WG, Capua I. A Closer Look at the NS1 of Influenza Virus. Viruses 2009 Dec;1(3):1057-72.
- Kim MC, Choi JG, Kwon JS, Kang HM, Paek MR, Jeong OM, Kwon JH, Lee YJ. Field application of the H9M2e enzyme-linked immunosorbent assay for differentiation of H9N2 avian influenza virus-infected chickens from vaccinated chickens. Clin Vaccine Immunol 2010 Dec;17(12):1977-84.
- Krejnusová I, Gocníková H, Bystrická M, Blaskovicová H, Poláková K, Yewdell J, Bennink J, Russ G. Antibodies to PB1-F2 protein are induced in response to influenza A virus infection. Arch Virol 2009;154(10):1599-604.
- Yu CI, Gallegos M, Marches F, Zurawski G, Ramilo O, García-Sastre A, Banchereau J, Palucka AK. Broad influenza-specific CD8+ T-cell responses in humanized mice vaccinated with influenza virus vaccines. Blood 2008 Nov 1;112(9):3671-8.
- Tumpey TM, Alvarez R, Swayne DE, Suarez DL. Diagnostic approach for differentiating infected from vaccinated poultry on the basis of antibodies to NS1, the nonstructural protein of influenza A virus. J Clin Microbiol 2005 Feb;43(2):676-83.
- Ferko B, Stasakova J, Sereinig S, Romanova J, Katinger D, Niebler B, Katinger H, Egorov A. Hyperattenuated recombinant influenza A virus nonstructural-protein-encoding vectors induce human immunodeficiency virus type 1 Nef-specific systemic and mucosal immune responses in mice. J Virol 2001 Oct;75(19):8899-908.
- Gonzalez-Obando J, Jaimes-Dueñez J, Zuluaga-Cabrera A, Forero JE, Diaz A, Rojas-Arbeláez C, Ruiz-Saenz J. Seroprevalence of Equine Influenza Virus Antibodies in Horses from Four Localities in Colombia. Viruses 2025 Jul 16;17(7).
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