Broader Neutralizing Antibodies against H5N1 Viruses Using Prime-Boost Immunization of Hyperglycosylated Hemagglutinin DNA and Virus-Like Particles

Background Highly pathogenic avian influenza (HPAI) H5N1 viruses and their transmission capability from birds to humans have raised global concerns about a potential human pandemic. The inherent nature of antigenic changes in influenza viruses has not been sufficiently taken into account in immunogen designs for broadly protective HPAI H5N1 vaccines. Methods We designed a hyperglycosylated HA vaccine using N-linked glycan masking on highly variable sequences in the HA1 globular head. Immunization of these hyperglycosylated HA DNA vaccines followed by a flagellin-containing virus-like particle booster in mice was conducted to evaluate neutralizing antibody responses against various clades of HPAI H5N1 viruses. Results We introduced nine N-X-S/T motifs in five HA1 regions: 83NNT, 86NNT, 94NFT, 127NSS, 138NRT, 156NTT, 161NRS, 182NDT, and 252NAT according to sequence alignment analyses from 163 HPAI H5N1 human isolates. Although no significant differences of anti-HA total IgG titers were found with these hyperglycosyalted HA compared to the wild-type control, the 83NNT and 127NSS mutants elicited significantly potent cross-clade neutralizing antibodies against HPAI H5N1 viruses. Conclusions This finding may have value in terms of novel immunogen design for developing cross-protective H5N1 vaccines.

• Publication year

  2012

• Venue

  PLoS ONE

• Publication date

  2012-06-13

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1371/journal.pone.0039075PMID 22720032PMCID 3374787
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Naturally Occurring Antibodies in Humans Can Neutralize a Variety of Influenza Virus Strains, Including H3, H1, H2, and H5

  2011cited by this paper
• Recombinant Trimeric HA Protein Immunogenicity of H5N1 Avian Influenza Viruses and Their Combined Use with Inactivated or Adenovirus Vaccines

  2011cited by this paper
• A Neutralizing Antibody Selected from Plasma Cells That Binds to Group 1 and Group 2 Influenza A Hemagglutinins

  2011cited by this paper
• Superior Neutralizing Antibody Response and Protection in Mice Vaccinated with Heterologous DNA Prime and Virus Like Particle Boost against HPAI H5N1 Virus

  2011cited by this paper
• A Broadly Neutralizing Human Monoclonal Antibody That Recognizes a Conserved, Novel Epitope on the Globular Head of the Influenza H1N1 Virus Hemagglutinin

  2011cited by this paper
• Fabrication of influenza virus-like particles using M2 fusion proteins for imaging single viruses and designing vaccines.

  2011cited by this paper
• Influenza H5 Hemagglutinin DNA Primes the Antibody Response Elicited by the Live Attenuated Influenza A/Vietnam/1203/2004 Vaccine in Ferrets

  2011cited by this paper
• DNA priming and influenza vaccine immunogenicity: two phase 1 open label randomised clinical trials

  2011cited by this paper
• Broadly neutralizing human antibody that recognizes the receptor-binding pocket of influenza virus hemagglutinin

  2011cited by this paper
• Influenza Virus Vaccine Based on the Conserved Hemagglutinin Stalk Domain

  2010cited by this paper
• Emulsified Nanoparticles Containing Inactivated Influenza Virus and CpG Oligodeoxynucleotides Critically Influences the Host Immune Responses in Mice

  2010cited by this paper
• Structure–function relationships of HIV-1 envelope sequence-variable regions refocus vaccine design

  2010cited by this paper
• Structures of Receptor Complexes of a North American H7N2 Influenza Hemagglutinin with a Loop Deletion in the Receptor Binding Site

  2010cited by this paper
• Rational antibody-based HIV-1 vaccine design: current approaches and future directions.

  2010cited by this paper
• How Can Vaccines Against Influenza and Other Viral Diseases Be Made More Effective?

  2010cited by this paper
• Structural and functional bases for broad-spectrum neutralization of avian and human influenza A viruses

  2009cited by this paper
• Adjuvants for pandemic influenza vaccines.

  2009cited by this paper
• HIV-1 and influenza antibodies: seeing antigens in new ways

  2009cited by this paper
• Cross-clade neutralizing antibodies against HIV-1 induced in rabbits by focusing the immune response on a neutralizing epitope.

  2009cited by this paper
• Focusing the immune response on the V3 loop, a neutralizing epitope of the HIV-1 gp120 envelope.

  2008cited by this paper
• Heterologous HA DNA vaccine prime--inactivated influenza vaccine boost is more effective than using DNA or inactivated vaccine alone in eliciting antibody responses against H1 or H3 serotype influenza viruses.

  2008cited by this paper
• The challenges of eliciting neutralizing antibodies to HIV-1 and to influenza virus

  2008cited by this paper
• Improved Induction of Antibodies against Key Neutralizing Epitopes by Human Immunodeficiency Virus Type 1 gp120 DNA Prime-Protein Boost Vaccination Compared to gp120 Protein-Only Vaccination

  2008cited by this paper
• Focused Dampening of Antibody Response to the Immunodominant Variable Loops by Engineered Soluble gp140

  2008cited by this paper
• DNA vaccines: ready for prime time?

  2008cited by this paper
• Deceptive imprinting and immune refocusing in vaccine design.

  2008cited by this paper
• Comparison of vaccines for induction of heterosubtypic immunity to influenza A virus: cold-adapted vaccine versus DNA prime-adenovirus boost strategies.

  2008cited by this paper
• Immunization by Avian H5 Influenza Hemagglutinin Mutants with Altered Receptor Binding Specificity

  2007cited by this paper
• Comparing Antigenicity and Immunogenicity of Engineered gp120

  2005cited by this paper
• Hyperglycosylated Mutants of Human Immunodeficiency Virus (HIV) Type 1 Monomeric gp120 as Novel Antigens for HIV Vaccine Design

  2003cited by this paper
• Induction of Macrophage Nitric Oxide Production by Gram-Negative Flagellin Involves Signaling Via Heteromeric Toll-Like Receptor 5/Toll-Like Receptor 4 Complexes1

  2003cited by this paper
• Refocusing of B‐cell responses following a single amino acid substitution in an antigen

  2001cited by this paper
• Activation of Interleukin-1 Receptor-Associated Kinase by Gram-Negative Flagellin

  2001cited by this paper
• Studies of the Neutralizing Activity and Avidity of Anti-Human Immunodeficiency Virus Type 1 Env Antibody Elicited by DNA Priming and Protein Boosting

  1998cited by this paper
• Can epitope-focused vaccines select advantageous immune responses?

  1997cited by this paper
• Refocusing neutralizing antibody response by targeted dampening of an immunodominant epitope.

  1997cited by this paper
• Vaccination with HIV-1 gp120 DNA induces immune responses that are boosted by a recombinant gp120 protein subunit.

  1997cited by this paper
• The antigenic structure of the influenza virus A/PR/8/34 hemagglutinin (H1 subtype).

  1982cited by this paper
• Antigenic structure of influenza virus haemagglutinin defined by hybridoma antibodies

  1981cited by this paper

• Architects of folding, editors of immunity: the strategic use of N-glycans in vaccine design.

  2026cites this paper
• Glycan masking in immunogen design: computational and experimental methods

  2026cites this paper
• A consensus HA DNA vaccine targeting clade 2.3.4.4 H5Nx influenza viruses provides broad cross-clade protection in mice and ferrets.

  2025cites this paper
• Impact of glycosylation on viral vaccines.

  2024cites this paper
• Design of universal Ebola virus vaccine candidates via immunofocusing

  2024cites this paper
• Bringing immunofocusing into focus

  2024cites this paper
• Vaccine design via antigen reorientation

  2024cites this paper
• Prokaryote- and Eukaryote-Based Expression Systems: Advances in Post-Pandemic Viral Antigen Production for Vaccines

  2024cites this paper
• Glycan masking of NTD loops with a chimeric RBD of the spike protein as a vaccine design strategy against emerging SARS‐CoV‐2 Omicron variants

  2024cites this paper
• Structural insights into the broad protection against H1 influenza viruses by a computationally optimized hemagglutinin vaccine

  2023cites this paper
• Inducing enhanced neutralizing antibodies against broad SARS-CoV-2 variants through glycan-shielding multiple non-neutralizing epitopes of RBD

  2023cites this paper
• Design of universal Ebola virus vaccine candidates via immunofocusing

  2023cites this paper
• Glycovaccinology: The design and engineering of carbohydrate-based vaccine components.

  2023cites this paper
• Influenza H3 hemagglutinin vaccine with scrambled immunodominant epitopes elicits antibodies directed toward immunosubdominant head epitopes

  2023cites this paper
• Glycan masking in vaccine design: Targets, immunogens and applications

  2023cites this paper
• SARS-CoV-2 epitopes inform future vaccination strategies

  2022cites this paper
• Designing epitope-focused vaccines via antigen reorientation

  2022cites this paper
• Status and Challenges for Vaccination against Avian H9N2 Influenza Virus in China

  2022cites this paper
• Glycan Masking of Epitopes in the NTD and RBD of the Spike Protein Elicits Broadly Neutralizing Antibodies Against SARS-CoV-2 Variants

  2021cites this paper
• Adenoviral Vectors as Vaccines for Emerging Avian Influenza Viruses

  2021cites this paper
• Ebola-GP DNA Prime rAd5-GP Boost: Influence of Prime Frequency and Prime/Boost Time Interval on the Immune Response in Non-human Primates

  2021cites this paper
• Strategies Targeting Hemagglutinin as a Universal Influenza Vaccine

  2021cites this paper
• SARS-CoV-2 cell entry and targeted antiviral development

  2021cites this paper
• Targeting Antigens for Universal Influenza Vaccine Development

  2021cites this paper
• Site-Specific Glycan-Masking/Unmasking Hemagglutinin Antigen Design to Elicit Broadly Neutralizing and Stem-Binding Antibodies Against Highly Pathogenic Avian Influenza H5N1 Virus Infections

  2021cites this paper
• Targeting Viral Surface Proteins through Structure-Based Design

  2021cites this paper
• Influenza vaccine: progress in a vaccine that elicits a broad immune response

  2021cites this paper
• Glycan-masking spike antigen in NTD and RBD elicits broadly neutralizing antibodies against SARS-CoV-2 variants

  2021cites this paper
• Flu Universal Vaccines: New Tricks on an Old Virus

  2020cites this paper
• Glycomics and glycoproteomics of viruses: Mass spectrometry applications and insights toward structure–function relationships

  2020cites this paper
• Glycosylation as a tool for rational vaccine design

  2020cites this paper
• Development of broadly reactive influenza vaccines by targeting the conserved regions of the hemagglutinin stem and head domains

  2020cites this paper
• Call for a paradigm shift in the design of universal influenza vaccines by harnessing multiple correlates of protection

  2020cites this paper
• Human COBRA 2 vaccine contains two major epitopes that are responsible for eliciting neutralizing antibody responses against heterologous clades of viruses.

  2019cites this paper
• The Quest for a Truly Universal Influenza Vaccine

  2019cites this paper
• Is It Possible to Develop a "Universal" Influenza Virus Vaccine? Potential for a Universal Influenza Vaccine.

  2018cites this paper
• Mini-HA Is Superior to Full Length Hemagglutinin Immunization in Inducing Stem-Specific Antibodies and Protection Against Group 1 Influenza Virus Challenges in Mice

  2018cites this paper
• Glycan‐masking hemagglutinin antigens from stable CHO cell clones for H5N1 avian influenza vaccine development

  2018cites this paper
• Mini-hemagglutinin vaccination induces cross-reactive antibodies in pre-exposed NHP that protect mice against lethal influenza challenge

  2018cites this paper
• Protect, modify, deprotect (PMD): A strategy for creating vaccines to elicit antibodies targeting a specific epitope

  2018cites this paper
• A prime/boost vaccination with HA DNA and Pichia-produced HA protein elicits a strong humoral response in chickens against H5N1.

  2017cites this paper
• Strategies to induce broadly protective antibody responses to viral glycoproteins

  2017cites this paper
• Unmasking Stem-Specific Neutralizing Epitopes by Abolishing N-Linked Glycosylation Sites of Influenza Virus Hemagglutinin Proteins for Vaccine Design

  2016cites this paper
• Characterization of Humoral Responses Induced by an H7N9 Influenza Virus-Like Particle Vaccine in BALB/C Mice

  2015cites this paper
• Ferret models of viral pathogenesis

  2015cites this paper
• Prospects for antibody-based universal influenza vaccines in the context of widespread pre-existing immunity

  2015cites this paper
• Advances in the development of influenza virus vaccines

  2015cites this paper
• Protection against Multiple Influenza A Virus Strains Induced by Candidate Recombinant Vaccine Based on Heterologous M2e Peptides Linked to Flagellin

  2015cites this paper
• Glycan Masking of Hemagglutinin for Adenovirus Vector and Recombinant Protein Immunizations Elicits Broadly Neutralizing Antibodies against H5N1 Avian Influenza Viruses

  2014cites this paper
• DNA vaccines against influenza.

  2014cites this paper
• InfluenzaAntibody-Mediated Protection against MHC Class II Molecules Rapidly Induces DNA Vaccine that Targets Hemagglutinin to

  2013cites this paper
• Heterologous prime-boost immunization regimens using adenovirus vector and virus-like particles induce broadly neutralizing antibodies against H5N1 avian influenza viruses.

  2013cites this paper
• Different Immunity Elicited by Recombinant H5N1 Hemagglutinin Proteins Containing Pauci-Mannose, High-Mannose, or Complex Type N-Glycans

  2013cites this paper
• Beak and feather disease virus candidate vaccine development

  2012cites this paper