scholarly journals 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 ◽  
Vol 13 (9) ◽  
pp. 1708
Author(s):  
Nutan Mytle ◽  
Sonja Leyrer ◽  
Jon R. Inglefield ◽  
Andrea M. Harris ◽  
Thomas E. Hickey ◽  
...  
Keyword(s):  
Influenza A ◽  
Matrix Protein ◽  
Tandem Repeats ◽  
Protective Antigen ◽  
Serum Antibody ◽  
Cross Protection ◽  
Antigenic Drift ◽  
Natural Immunity ◽  
Pandemic H1n1 ◽  
H5n1 Influenza

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.

scholarly journals Serum Antibody Response to Matrix Protein 2 Following Natural Infection With 2009 Pandemic Influenza A(H1N1) Virus in Humans

2013 ◽  
Vol 209 (7) ◽  
pp. 986-994 ◽  
Author(s):  
Weimin Zhong ◽  
Carrie Reed ◽  
Patrick J. Blair ◽  
Jacqueline M. Katz ◽  
Kathy Hancock ◽  
...  
Keyword(s):  
Antibody Response ◽  
Pandemic Influenza ◽  
Influenza A ◽  
Matrix Protein ◽  
H1n1 Virus ◽  
Natural Infection ◽  
Serum Antibody ◽  
Serum Antibody Response ◽  
Influenza A H1n1

scholarly journals Matrix Protein 2 Extracellular Domain-Specific Monoclonal Antibodies Are an Effective and Potentially Universal Treatment for Influenza A

2020 ◽  
Author(s):  
Lynn Bimler ◽  
Sydney L. Ronzulli ◽  
Amber Y. Song ◽  
Scott K. Johnson ◽  
Cheryl A. Jones ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Monoclonal Antibodies ◽  
Human Health ◽  
Influenza A ◽  
Matrix Protein ◽  
Critical Role ◽  
Extracellular Domain ◽  
Influenza Viruses ◽  
Antigenic Drift ◽  
Protective Potential

Influenza virus infection causes significant morbidity and mortality worldwide. Humans fail to make a universally protective memory immune response to influenza A. Hemagglutinin and Neuraminidase undergo antigenic drift and shift, resulting in new influenza A strains to which humans are naive. Seasonal vaccines are often ineffective and escape mutants have been reported to all treatments for influenza A. In the absence of a universal influenza A vaccine or treatment, influenza A will remain a significant threat to human health. The extracellular domain of the M2-ion channel (M2e) is an ideal antigenic target for a universal therapeutic agent, as it is highly conserved across influenza A serotypes, has a low mutation rate, and is essential for viral entry and replication. Previous M2e-specific monoclonal antibodies (M2e-MAbs) show protective potential against influenza A, however, they are either strain specific or have limited efficacy. We generated seven murine M2e-MAbs and utilized in vitro and in vivo assays to validate the specificity of our novel M2e-MAbs and to explore the universality of their protective potential. Our data shows our M2e-MAbs bind to M2e peptide, HEK cells expressing the M2 channel, as well as, influenza virions and MDCK-ATL cells infected with influenza viruses of multiple serotypes. Our antibodies significantly protect highly influenza A virus susceptible BALB/c mice from lethal challenge with H1N1 A/PR/8/34, pH1N1 A/CA/07/2009, H5N1 A/Vietnam/1203/2004, and H7N9 A/Anhui/1/2013 by improving survival rates and weight loss. Based on these results, at least four of our seven M2e-MAbs show strong potential as universal influenza A treatments. IMPORTANCE Despite a seasonal vaccine and multiple therapeutic treatments, Influenza A remains a significant threat to human health. The biggest obstacle is producing a vaccine or treatment for influenza A is their universality or efficacy against not only seasonal variances in the influenza virus, but also against all human, avian, and swine serotypes and, therefore, potential pandemic strains. M2e has huge potential as a target for a vaccine or treatment against influenza A. It is the most conserved external protein on the virus. Antibodies against M2e have made it to clinical trials, but not succeeded. Here, we describe novel M2e antibodies produced in mice that are not only protective at low doses, but that we extensively test to determine their universality and found to be cross protective against all strains tested. Additionally, our work begins to elucidate the critical role of isotype for an influenza A monoclonal antibody therapeutic.

scholarly journals Immunogenicity and Protective Potential of Mucosal Vaccine Formulations Based on Conserved Epitopes of Influenza A Viruses Fused to an Innovative Ring Nanoplatform in Mice and Chickens

2021 ◽  
Vol 12 ◽  
Author(s):  
Cynthia Calzas ◽  
Molida Mao ◽  
Mathilde Turpaud ◽  
Quentin Viboud ◽  
Joelle Mettier ◽  
...  
Keyword(s):  
Influenza A ◽  
Matrix Protein ◽  
Tandem Repeats ◽  
Alpha Helix ◽  
Humoral Response ◽  
Mucosal Vaccine ◽  
Influenza A Viruses ◽  
Protective Potential ◽  
Syncytial Virus

Current inactivated vaccines against influenza A viruses (IAV) mainly induce immune responses against highly variable epitopes across strains and are mostly delivered parenterally, limiting the development of an effective mucosal immunity. In this study, we evaluated the potential of intranasal formulations incorporating conserved IAV epitopes, namely the long alpha helix (LAH) of the stalk domain of hemagglutinin and three tandem repeats of the ectodomain of the matrix protein 2 (3M2e), as universal mucosal anti-IAV vaccines in mice and chickens. The IAV epitopes were grafted to nanorings, a novel platform technology for mucosal vaccination formed by the nucleoprotein (N) of the respiratory syncytial virus, in fusion or not with the C-terminal end of the P97 protein (P97c), a recently identified Toll-like receptor 5 agonist. Fusion of LAH to nanorings boosted the generation of LAH-specific systemic and local antibody responses as well as cellular immunity in mice, whereas the carrier effect of nanorings was less pronounced towards 3M2e. Mice vaccinated with chimeric nanorings bearing IAV epitopes in fusion with P97c presented modest LAH- or M2e-specific IgG titers in serum and were unable to generate a mucosal humoral response. In contrast, N-3M2e or N-LAH nanorings admixed with Montanide™ gel (MG) triggered strong specific humoral responses, composed of serum type 1/type 2 IgG and mucosal IgG and IgA, as well as cellular responses dominated by type 1/type 17 cytokine profiles. All mice vaccinated with the [N-3M2e + N-LAH + MG] formulation survived an H1N1 challenge and the combination of both N-3M2e and N-LAH nanorings with MG enhanced the clinical and/or virological protective potential of the preparation in comparison to individual nanorings. Chickens vaccinated parenterally or mucosally with N-LAH and N-3M2e nanorings admixed with Montanide™ adjuvants developed a specific systemic humoral response, which nonetheless failed to confer protection against heterosubtypic challenge with a highly pathogenic H5N8 strain. Thus, while the combination of N-LAH and N-3M2e nanorings with Montanide™ adjuvants shows promise as a universal mucosal anti-IAV vaccine in the mouse model, further experiments have to be conducted to extend its efficacy to poultry.

scholarly journals Production of Influenza Virus Proteins Using Recombinant Insect Cells

2021 ◽  
Vol 333 ◽  
pp. 07009
Author(s):  
Takuya Matsuda ◽  
Toshikazu Tanijima ◽  
Kyoko Masumi-Koizumi ◽  
Tomohisa Katsuda ◽  
Hideki Yamaji
Keyword(s):  
Influenza Virus ◽  
Influenza A ◽  
Matrix Protein ◽  
Insect Cells ◽  
Protective Antigen ◽  
Continuous Production ◽  
Recombinant Baculovirus ◽  
Cell System ◽  
Structural Protein ◽  
Production Period

Influenza vaccines have long been manufactured in embryonated chicken eggs. This method has some problems such as a long production period (about 6 months) and use of large amounts of infectious pathogens. Recently, the production of recombinant subunit vaccines using the baculovirus–insect cell system has been extensively investigated. In this system, viral immunodominant components can be produced more rapidly and in a larger scale than in the conventional egg-based process. However, continuous production is virtually impossible because infection of recombinant baculovirus results in the death of host insect cells. In the present study, we established stably transformed insect cells that secreted influenza virus-like particles (VLPs) consisting of hemagglutinin (HA), the major protective antigen of influenza A virus, and matrix protein 1 (M1), another structural protein of the virus. Hemagglutination assay and transmission electron microscopy (TEM) suggested that HA produced by recombinant insect cells kept the hemagglutination activity and the morphology of the VLPs was similar to that of wild type influenza virus particles.

scholarly journals Identification of mammalian-adapting mutations involved into antigenic change of H1N1 influenza virus

2020 ◽  
Author(s):  
Zhijun Yu ◽  
Kaihui Cheng ◽  
Weiyang Sun ◽  
Xinghai Zhang ◽  
Tiecheng Wang ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Amino Acid ◽  
Influenza A ◽  
H1n1 Virus ◽  
Serial Passage ◽  
Antigenic Drift ◽  
Amino Acid Substitutions ◽  
Pandemic H1n1 ◽  
Potential Threat ◽  
Ha Protein

Abstract Though the 2009 pandemic H1N1 virus has become a seasonal influenza A virus, continued host adaptation and antigenic drift may limit the effectiveness of current vaccines. Here, we assess viral antigenic change of a 2009 pandemic H1N1 virus (A/Changchun/01/2009) in the absence and presence of immune-mediated selective pressures and further identify the key amino acid substitutions involved into the antigenic change of the 2009 pandemic H1N1 virus. We found that serial passage of the 2009 pandemic H1N1 virus in both naïve and previously immunized mice generated antigenically distinct variants and that serial passage in previously immunized mice generated viruses with lower overall antigenic relatedness to the parental virus when compared to variants generated by serial passage in naïve mice, suggesting that antibody pressure may accelerate the antigenic variation rate of the 2009 pandemic H1N1 virus. Furthermore, we found that three amino acid substitutions in the viral HA protein (N159D, S186P, and D225G), both alone and in combination, affected viral antigenicity. It is noted that all three mutations (N159D, S186P, and D225G) in the viral HA protein have been found in the natural 2009 pandemic H1N1 virus isolates and can also enhance the pathogenicity of the 2009 pandemic H1N1 virus to mammals, suggesting that these mutations in the viral HA protein may pose a potential threat to public health and should be paid more attention. Taken together, our work defines some novel molecular determinants of the pandemic H1N1 virus antigenicity and has important implications for ongoing human influenza virus surveillance efforts.

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