scholarly journals Protection of chickens against A/H5N1 virus by plant-based haemagglutinin fused IGMFC

2020 ◽  
Vol 18 (3) ◽  
pp. 477-485
Author(s):  
Pham Thi Van ◽  
Ho Thi Huong ◽  
Nguyen Thu Giang ◽  
Pham Bich Ngoc ◽  
Vu Huyen Trang ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Neutralizing Antibodies ◽  
H5n1 Virus ◽  
Positive Control ◽  
Joint Stock Company ◽  
Stock Company ◽  
H5n1 Avian Influenza Virus ◽  
Transmembrane Glycoprotein ◽  
H5n1 Influenza ◽  
Protection Rate

Vaccination is one of the most effective and cost-beneficial interventions for protection of animals against the highly pathogenic A/H5N1 avian influenza virus. Haemagglutinin (HA) is a transmembrane glycoprotein of A/H5N1 virus and is a critical antigen for development of the influenza vaccine. The haemagglutinin-based vaccine produced in plants was demonstrated as a candidate vaccine since it elicited neutralizing antibodies against A/H5N1 virus. In this study, immunogenicity and protective ability of a plant-based recombinant HA antigen which was fused to IgMFc to form oligomerized HA antigen (H5TG oligomer) had been evaluated by vaccination in chickens. Chicken sera after each vaccination were collected for Western blot, ELISA and HI assays. Ten days after the second vaccination, the chickens have been challenged with A/duck/TG/NAVET(3)/2013 virus, clade 1.1. The analysis results showed that the oligomeric recombinant H5TG antigen elicited stronger H5TG-specific IgY antibodies and A/H5N1 clade 1.1 virus-neutralizing antibodies than the H5TGpII trimeric recombinant antigen without fusing IgMFc in vaccinated chickens. Notably, the chicken protection rate against A/H5N1 clade 1.1 virus of the H5TG oligomer antigen was 80% that is not significantly lower than that of a commercial vaccine as a positive control from National Veterinary Joint Stock company NAVETCO, Vietnam with the chicken protective rate of 90%. Whereas the chicken protection rate against A/H5N1 clade 1.1 virus of the H5TG trimer antigen was 50%. These results suggest that the IgMFc motif plays an important role in the forming of oligomeric proteins which had been proved for enhancing immunogenicity and protection ability in this study. Therefore, the plant-based oligomerized recombinant H5TG antigen is a potential vaccine candidate against A/H5N1 influenza virus in the future.

Peptide mimics of a conserved H5N1 avian influenza virus neutralization site

2009 ◽  
Vol 419 (1) ◽  
pp. 133-139 ◽  
Author(s):  
Wenxin Luo ◽  
Yingwei Chen ◽  
Mingqiao Wang ◽  
Yixin Chen ◽  
Zhenhua Zheng ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
Fusion Proteins ◽  
H5n1 Virus ◽  
Amino Acid Sequences ◽  
Antigenic Determinants ◽  
H5n1 Avian Influenza Virus ◽  
H5n1 Influenza ◽  
H5n1 Avian Influenza

A panel of 52 murine monoclonal antibodies was found to recognize antigenic determinants that had been conserved among all major genetic subgroups of the H5N1 avian influenza virus prevalent since 1997. We screened a phage display library for peptides recognized by one such antibody (8H5). We analysed the specificity of 8H5 for reactive peptides presented as fusion proteins of HBc (hepatitis B core protein) and HEV (hepatitis E virus) structural protein, p239. This was then related to the specificity of the native HA (haemagglutinin) molecule by virtue of the capacity of fusion proteins to compete for 8H5 binding with different strains of H5N1 virus and the reactivity of antisera generated against fusion proteins to bind native HA molecules, and to inhibit haemagglutination and arrest infection by the virus. Nine reactive peptides of different amino acid sequences were identified, six of which were also reactive with the antibody in association with HBc and four were in association with p239. Binding occurred with the dimeric form of the four p239-fusion proteins and one of the HBc-fusion proteins, but not with the monomeric form. The HBc-fusion proteins blocked 8H5 binding with four strains of H5N1 influenza virus. Mouse antisera generated against fusion proteins bound to HA molecules, but did not inhibit haemagglutination or arrest H5N1 infection. Our findings indicate that 8H5 recognizes discontinuous sites presented by secondary and possibly higher structural orders of the peptides in spatially favourable positions for binding with the antibody, and that the peptides partially mimic the native 8H5 epitopes on the H5N1 virus.

scholarly journals H5N1 Influenza Virus Pathogenesis in Genetically Diverse Mice Is Mediated at the Level of Viral Load

mBio ◽  
2011 ◽  
Vol 2 (5) ◽  
Author(s):  
Adrianus C. M. Boon ◽  
David Finkelstein ◽  
Ming Zheng ◽  
Guochun Liao ◽  
John Allard ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Viral Load ◽  
H5n1 Virus ◽  
Inbred Mouse ◽  
Severe Disease ◽  
Mouse Strains ◽  
H5n1 Influenza Virus ◽  
Resistant Mouse ◽  
H5n1 Influenza ◽  
Host Genetic

ABSTRACTThe genotype of the host is one of several factors involved in the pathogenesis of an infectious disease and may be a key parameter in the epidemiology of highly pathogenic H5N1 influenza virus infection in humans. Gene polymorphisms may affect the viral replication rate or alter the host’s immune response to the virus. In humans, it is unclear which aspect dictates the severity of H5N1 virus disease. To identify the mechanism underlying differential responses to H5N1 virus infection in a genetically diverse population, we assessed the host responses and lung viral loads in 21 inbred mouse strains upon intranasal inoculation with A/Hong Kong/213/03 (H5N1). Resistant mouse strains survived large inocula while susceptible strains succumbed to infection with 1,000- to 10,000-fold-lower doses. Quantitative analysis of the viral load after inoculation with an intermediate dose found significant associations with lethality as early as 2 days postinoculation, earlier than any other disease indicator. The increased viral titers in the highly susceptible strains mediated a hyperinflamed environment, indicated by the distinct expression profiles and increased production of inflammatory mediators on day 3. Supporting the hypothesis that viral load rather than an inappropriate response to the virus was the key severity-determining factor, we performed quantitative real-time PCR measuring the cytokine/viral RNA ratio. No significant differences between susceptible and resistant mouse strains were detected, confirming that it is the host genetic component controlling viral load, and therefore replication dynamics, that is primarily responsible for a host’s susceptibility to a given H5N1 virus.IMPORTANCEHighly pathogenic H5N1 influenza virus has circulated in Southeast Asia since 2003 but has been confirmed in relatively few individuals. It has been postulated that host genetic polymorphisms increase the susceptibility to infection and severe disease. The mechanisms and host proteins affected during severe disease are unknown. Inbred mouse strains vary considerably in their ability to resist H5N1 virus and were used to identify the primary mechanism determining disease severity. After inoculation with H5N1, resistant mouse strains had reduced amounts of virus in their lungs, which subsequently resulted in lower production of proinflammatory mediators and less pathology. We therefore conclude that the host genetic component controlling disease severity is primarily influencing viral replication. This is an important concept, as it emphasizes the need to limit virus replication through antiviral therapies and it shows that the hyperinflammatory environment is simply a reflection of more viral genetic material inducing a response.

scholarly journals An Antibody against a Novel and Conserved Epitope in the Hemagglutinin 1 Subunit Neutralizes Numerous H5N1 Influenza Viruses

2010 ◽  
Vol 84 (16) ◽  
pp. 8275-8286 ◽  
Author(s):  
Hsueh-Ling Janice Oh ◽  
Sara Åkerström ◽  
Shuo Shen ◽  
Sándor Bereczky ◽  
Helen Karlberg ◽  
...  
Keyword(s):  
Influenza A ◽  
H5n1 Virus ◽  
Influenza Viruses ◽  
Neutralization Titer ◽  
H5n1 Avian Influenza Virus ◽  
H5n1 Influenza ◽  
Neutralizing Monoclonal Antibody ◽  
Homologous Strain ◽  
Clade 1 ◽  
Conserved Epitope

ABSTRACT The spread of the recently emerged, highly pathogenic H5N1 avian influenza virus has raised concern. Preclinical studies suggest that passive immunotherapy could be a new form of treatment for H5N1 virus infection. Here, a neutralizing monoclonal antibody (MAb) against the hemagglutinin (HA) of the influenza A/chicken/Hatay/2004 H5N1 virus, MAb 9F4, was generated and characterized. MAb 9F4 binds both the denatured and native forms of HA. It was shown to recognize the HA proteins of three heterologous strains of H5N1 viruses belonging to clades 1, 2.1, and 2.2, respectively. By use of lentiviral pseudotyped particles carrying HA on the surface, MAb 9F4 was shown to effectively neutralize the homologous strain, Hatay04, and another clade 1 strain, VN04, at a neutralization titer of 8 ng/ml. Furthermore, MAb 9F4 also neutralized two clade 2 viruses at a neutralizing titer of 40 ng/ml. The broad cross-neutralizing activity of MAb 9F4 was confirmed by its ability to neutralize live H5N1 viruses of clade 2.2.2. Epitope-mapping analysis revealed that MAb 9F4 binds a previously uncharacterized epitope below the globular head of the HA1 subunit. Consistently, this epitope is well conserved among the different clades of H5N1 viruses. MAb 9F4 does not block the interaction between HA and its receptor but prevents the pH-mediated conformational change of HA. MAb 9F4 was also found to be protective, both prophylactically and therapeutically, against a lethal viral challenge of mice. Taken together, our results showed that MAb 9F4 is a neutralizing MAb that binds a novel and well-conserved epitope in the HA1 subunit of H5N1 viruses.

scholarly journals High Level of Genetic Compatibility between Swine-Origin H1N1 and Highly Pathogenic Avian H5N1 Influenza Viruses

2010 ◽  
Vol 84 (20) ◽  
pp. 10918-10922 ◽  
Author(s):  
Cássio Pontes Octaviani ◽  
Makoto Ozawa ◽  
Shinya Yamada ◽  
Hideo Goto ◽  
Yoshihiro Kawaoka
Keyword(s):  
Influenza Virus ◽  
H5n1 Virus ◽  
Human Lung ◽  
Cultured Cells ◽  
Influenza Viruses ◽  
Compatibility Studies ◽  
Genetic Compatibility ◽  
Highly Pathogenic ◽  
H5n1 Influenza ◽  
High Level

Reassortment is an important mechanism for the evolution of influenza viruses. Here, we coinfected cultured cells with the pandemic swine-origin influenza virus (S-OIV) and a contemporary H5N1 virus and found that these two viruses have high genetic compatibility. Studies of human lung cell lines indicated that some reassortants had better growth kinetics than their parental viruses. We conclude that reassortment between these two viruses can occur and could create pandemic H5N1 viruses.

scholarly journals Intranasal Vaccination with 1918 Influenza Virus-Like Particles Protects Mice and Ferrets from Lethal 1918 and H5N1 Influenza Virus Challenge

2009 ◽  
Vol 83 (11) ◽  
pp. 5726-5734 ◽  
Author(s):  
Lucy A. Perrone ◽  
Attiya Ahmad ◽  
Vic Veguilla ◽  
Xiuhua Lu ◽  
Gale Smith ◽  
...  
Keyword(s):  
Influenza Virus ◽  
H5n1 Virus ◽  
Influenza Pandemic ◽  
Human Case ◽  
Lethal Challenge ◽  
Virus Like Particles ◽  
Virus Challenge ◽  
H5n1 Influenza ◽  
Pandemic Virus ◽  
Homologous Virus

ABSTRACT Influenza vaccines capable of inducing cross-reactive or heterotypic immunity could be an important first line of prevention against a novel subtype virus. Influenza virus-like particles (VLPs) displaying functional viral proteins are effective vaccines against replication-competent homologous virus, but their ability to induce heterotypic immunity has not been adequately tested. To measure VLP vaccine efficacy against a known influenza pandemic virus, recombinant VLPs were generated from structural proteins of the 1918 H1N1 virus. Mucosal and traditional parenteral administrations of H1N1 VLPs were compared for the ability to protect against the reconstructed 1918 virus and a highly pathogenic avian H5N1 virus isolated from a fatal human case. Mice that received two intranasal immunizations of H1N1 VLPs were largely protected against a lethal challenge with both the 1918 virus and the H5N1 virus. In contrast, mice that received two intramuscular immunizations of 1918 VLPs were only protected against a homologous virus challenge. Mucosal vaccination of mice with 1918 VLPs induced higher levels of cross-reactive immunoglobulin G (IgG) and IgA antibodies than did parenteral vaccination. Similarly, ferrets mucosally vaccinated with 1918 VLPs completely survived a lethal challenge with the H5N1 virus, while only a 50% survival rate was observed in parenterally vaccinated animals. These results suggest a strategy of VLP vaccination against a pandemic virus and one that stimulates heterotypic immunity against an influenza virus strain with threatening pandemic potential.

scholarly journals Epitope Mapping of the Hemagglutinin Molecule of a Highly Pathogenic H5N1 Influenza Virus by Using Monoclonal Antibodies

2007 ◽  
Vol 81 (23) ◽  
pp. 12911-12917 ◽  
Author(s):  
Nikolai V. Kaverin ◽  
Irina A. Rudneva ◽  
Elena A. Govorkova ◽  
Tatyana A. Timofeeva ◽  
Aleksandr A. Shilov ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Amino Acid ◽  
H5n1 Virus ◽  
Antigenic Site ◽  
H5n1 Influenza Virus ◽  
Highly Pathogenic ◽  
H5n1 Influenza ◽  
Antigenic Sites ◽  
Escape Mutants ◽  
Pathogenic H5n1

ABSTRACT We mapped the hemagglutinin (HA) antigenic epitopes of a highly pathogenic H5N1 influenza virus on the three-dimensional HA structure by characterizing escape mutants of a recombinant virus containing A/Vietnam/1203/04 (H5N1) ΔHA and neuraminidase genes in the genetic background of A/Puerto Rico/8/34 (H1N1) virus. The mutants were selected with a panel of eight anti-HA monoclonal antibodies (MAbs), seven to A/Vietnam/1203/04 (H5N1) virus and one to A/Chicken/Pennsylvania/8125/83 (H5N2) virus, and the mutants’ HA genes were sequenced. The amino acid changes suggested three MAb groups: four MAbs reacted with the complex epitope comprising parts of the antigenic site B of H3 HA and site Sa of H1 HA, two MAbs reacted with the epitope corresponding to the antigenic site A in H3 HA, and two MAbs displayed unusual behavior: each recognized amino acid changes at two widely separate antigenic sites. Five changes were detected in amino acid residues not previously reported as changed in H5 escape mutants, and four others had substitutions not previously described. The HA antigenic structure differs substantially between A/Vietnam/1203/04 (H5N1) virus and the low-pathogenic A/Mallard/Pennsylvania/10218/84 (H5N2) virus we previously characterized (N. V. Kaverin et al., J. Gen. Virol. 83:2497-2505, 2002). The hemagglutination inhibition reactions of the MAbs with recent highly pathogenic H5N1 viruses were consistent with the antigenic-site amino acid changes but not with clades and subclades based on H5 phylogenetic analysis. These results provide information on the recognition sites of the MAbs widely used to study H5N1 viruses and demonstrate the involvement of the HA antigenic sites in the evolution of highly pathogenic H5N1 viruses, findings that can be critical for characterizing pathogenesis and vaccine design.

scholarly journals Complete Genomic Sequence of an H5N1 Influenza Virus from a Parrot in Southern China

2012 ◽  
Vol 86 (16) ◽  
pp. 8894-8895 ◽  
Author(s):  
Peirong Jiao ◽  
Yafen Song ◽  
Runyu Yuan ◽  
Liangmeng Wei ◽  
Lan Cao ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
Genomic Sequence ◽  
Southern China ◽  
H5n1 Avian Influenza Virus ◽  
H5n1 Influenza ◽  
H5n1 Avian Influenza ◽  
Eurasian Lineage ◽  
Complete Genomic

An H5N1 avian influenza virus (AIV) designated A/Parrot/Guangdong/C99/2005 (H5N1) was first isolated from a sick parrot in Guangdong in southern China in 2005. The complete genome of this strain was analyzed. Genome sequence analysis showed that all 8 gene segments of the virus nucleotide had 99.0% homology to A/chicken/Henan/12/2004 (H5N1). Phylogenetic analysis demonstrated that all 8 gene segments of the virus were derived from the Eurasian lineage. The availability of genome sequences is useful to investigate the host range and genetic evolution of the H5N1 avian influenza virus in Southern China.

scholarly journals A liposome-displayed hemagglutinin vaccine platform protects mice and ferrets from heterologous influenza virus challenge

2021 ◽  
Vol 118 (22) ◽  
pp. e2025759118
Author(s):  
Zachary R. Sia ◽  
Xuedan He ◽  
Ali Zhang ◽  
Jann C. Ang ◽  
Shuai Shao ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Neutralizing Antibodies ◽  
H3n2 Virus ◽  
Antigen Delivery ◽  
Vaccine Platform ◽  
Virus Challenge ◽  
H5n1 Influenza ◽  
Adjuvant System ◽  
Protein Particle

Recombinant influenza virus vaccines based on hemagglutinin (HA) hold the potential to accelerate production timelines and improve efficacy relative to traditional egg-based platforms. Here, we assess a vaccine adjuvant system comprised of immunogenic liposomes that spontaneously convert soluble antigens into a particle format, displayed on the bilayer surface. When trimeric H3 HA was presented on liposomes, antigen delivery to macrophages was improved in vitro, and strong functional antibody responses were induced following intramuscular immunization of mice. Protection was conferred against challenge with a heterologous strain of H3N2 virus, and naive mice were also protected following passive serum transfer. When admixed with the particle-forming liposomes, immunization reduced viral infection severity at vaccine doses as low as 2 ng HA, highlighting dose-sparing potential. In ferrets, immunization induced neutralizing antibodies that reduced the upper respiratory viral load upon challenge with a more modern, heterologous H3N2 viral strain. To demonstrate the flexibility and modular nature of the liposome system, 10 recombinant surface antigens representing distinct influenza virus strains were bound simultaneously to generate a highly multivalent protein particle that with 5 ng individual antigen dosing induced antibodies in mice that specifically recognized the constituent immunogens and conferred protection against heterologous H5N1 influenza virus challenge. Taken together, these results show that stable presentation of recombinant HA on immunogenic liposome surfaces in an arrayed fashion enhances functional immune responses and warrants further attention for the development of broadly protective influenza virus vaccines.

scholarly journals A Single Immunization with CoVaccine HT-Adjuvanted H5N1 Influenza Virus Vaccine Induces Protective Cellular and Humoral Immune Responses in Ferrets

2010 ◽  
Vol 84 (16) ◽  
pp. 7943-7952 ◽  
Author(s):  
R. Bodewes ◽  
J. H. C. M. Kreijtz ◽  
G. van Amerongen ◽  
M. M. Geelhoed-Mieras ◽  
R. J. Verburgh ◽  
...  
Keyword(s):  
Cell Culture ◽  
Influenza Virus ◽  
Immune Responses ◽  
Influenza A Virus ◽  
Virus Vaccine ◽  
Influenza A ◽  
H5n1 Virus ◽  
The Novel ◽  
Highly Pathogenic ◽  
H5n1 Influenza

ABSTRACT Highly pathogenic avian influenza A viruses of the H5N1 subtype continue to circulate in poultry, and zoonotic transmissions are reported frequently. Since a pandemic caused by these highly pathogenic viruses is still feared, there is interest in the development of influenza A/H5N1 virus vaccines that can protect humans against infection, preferably after a single vaccination with a low dose of antigen. Here we describe the induction of humoral and cellular immune responses in ferrets after vaccination with a cell culture-derived whole inactivated influenza A virus vaccine in combination with the novel adjuvant CoVaccine HT. The addition of CoVaccine HT to the influenza A virus vaccine increased antibody responses to homologous and heterologous influenza A/H5N1 viruses and increased virus-specific cell-mediated immune responses. Ferrets vaccinated once with a whole-virus equivalent of 3.8 μg hemagglutinin (HA) and CoVaccine HT were protected against homologous challenge infection with influenza virus A/VN/1194/04. Furthermore, ferrets vaccinated once with the same vaccine/adjuvant combination were partially protected against infection with a heterologous virus derived from clade 2.1 of H5N1 influenza viruses. Thus, the use of the novel adjuvant CoVaccine HT with cell culture-derived inactivated influenza A/H5N1 virus antigen is a promising and dose-sparing vaccine approach warranting further clinical evaluation.

scholarly journals Characterization of a highly pathogenic H5N1 influenza virus derived from bar-headed geese in China

2006 ◽  
Vol 87 (7) ◽  
pp. 1823-1833 ◽  
Author(s):  
Ji-Yong Zhou ◽  
Hui-Gang Shen ◽  
Hong-Xun Chen ◽  
Guang-Zhi Tong ◽  
Ming Liao ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Avian Influenza ◽  
Avian Influenza Virus ◽  
H5n1 Virus ◽  
H5n1 Influenza Virus ◽  
Highly Pathogenic ◽  
H5n1 Influenza ◽  
H5n1 Avian Influenza ◽  
Pathogenic H5n1 ◽  
Migratory Waterbirds

Influenza A viruses are usually non-pathogenic in wild aquatic birds, their natural reservoir. However, from May to July 2005, at Qinghai Lake in China, an unprecedented outbreak of highly pathogenic H5N1 avian influenza virus caused the death of thousands of wild migratory waterbirds. Herein, H5N1 influenza virus from bar-headed geese collected during the outbreak was characterized. Genomic analysis showed that A/Bar-headed Goose/Qinghai/0510/05 (Bh H5N1 virus) is a reassortant virus. Amino acid residue (lysine) at position 627 in the PB2 gene of the Bh H5N1 virus was the same as that of the human H5N1 virus (A/HK/483/97) and different from that of H5N1 avian influenza viruses deposited in GenBank. Antigenic analysis showed that significant antigenic variation has occurred in the Bh H5N1 virus. The Bh H5N1 virus induced systemic infections and caused 100 % mortality in chickens and mice, and 80 % mortality in ducks and geese. Bh H5N1 virus titres were higher in multiple organs of chickens, ducks and geese than in mice, and caused more severe histological lesions in chickens, ducks and mice than in geese. These results support the need to pay close attention and create control programmes to prevent the transmission of highly pathogenic avian influenza virus from wild migratory waterbirds into domestic chickens, ducks, geese and mammalian hosts.

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