Influenza Antigens NP and M2 Confer Cross Protection to BALB/c Mice against Lethal Challenge with H1N1, Pandemic H1N1 or H5N1 Influenza A Viruses

Viruses. 2021 Aug 27;13(9):1708. doi: 10.3390/v13091708.


Influenza hemagglutinin (HA) is considered a major protective antigen of seasonal influenza vaccine but antigenic drift of HA necessitates annual immunizations using new circulating HA versions. Low variation found within conserved non-HA influenza virus (INFV) antigens may maintain protection with less frequent immunizations. Conserved antigens of influenza A virus (INFV A) that can generate cross protection against multiple INFV strains were evaluated in BALB/c mice using modified Vaccinia virus Ankara (MVA)-vectored vaccines that expressed INFV A antigens hemagglutinin (HA), matrix protein 1 (M1), nucleoprotein (NP), matrix protein 2 (M2), repeats of the external portion of M2 (M2e) or as tandem repeats (METR), and M2e with transmembrane region and cytoplasmic loop (M2eTML). Protection by combinations of non-HA antigens was equivalent to that of subtype-matched HA. Combinations of NP and forms of M2e generated serum antibody responses and protected mice against lethal INFV A challenge using PR8, pandemic H1N1 A/Mexico/4108/2009 (pH1N1) or H5N1 A/Vietnam/1203/2004 (H5N1) viruses, as demonstrated by reduced lung viral burden and protection against weight loss. The highest levels of protection were obtained with NP and M2e antigens delivered as MVA inserts, resulting in broadly protective immunity in mice and enhancement of previous natural immunity to INFV A.

Keywords: conserved antigens; hemagglutinin; influenza A; matrix protein 1; matrix protein 2; modified Vaccinia virus Ankara (MVA); nucleoprotein; vaccine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral / blood*
  • Antigens, Viral / immunology
  • Cross Protection
  • Female
  • Genetic Vectors
  • Influenza A Virus, H1N1 Subtype / immunology*
  • Influenza A Virus, H1N1 Subtype / pathogenicity
  • Influenza A Virus, H5N1 Subtype / immunology*
  • Influenza A Virus, H5N1 Subtype / pathogenicity
  • Influenza Vaccines / administration & dosage
  • Influenza Vaccines / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Nucleocapsid Proteins / administration & dosage
  • Nucleocapsid Proteins / immunology*
  • Orthomyxoviridae Infections / immunology
  • Orthomyxoviridae Infections / prevention & control*
  • Pandemics
  • Vaccination
  • Viral Matrix Proteins / administration & dosage
  • Viral Matrix Proteins / genetics
  • Viral Matrix Proteins / immunology*
  • Viroporin Proteins / administration & dosage
  • Viroporin Proteins / immunology*


  • Antibodies, Viral
  • Antigens, Viral
  • Influenza Vaccines
  • M2 protein, Influenza A virus
  • NP protein, Influenza A virus
  • Nucleocapsid Proteins
  • Viral Matrix Proteins
  • Viroporin Proteins