Enhanced Immunogenicity of Stabilized Trimeric Soluble Influenza Hemagglutinin

Background The recent swine-origin H1N1 pandemic illustrates the need to develop improved procedures for rapid production of influenza vaccines. One alternative to the current egg-based manufacture of influenza vaccine is to produce a hemagglutinin (HA) subunit vaccine using a recombinant expression system with the potential for high protein yields, ease of cloning new antigenic variants, and an established safety record in humans. Methodology/Principal Findings We generated a soluble HA (sHA), derived from the H3N2 virus A/Aichi/2/68, modified at the C-terminus with a GCN4pII trimerization repeat to stabilize the native trimeric structure of HA. When expressed in the baculovirus system, the modified sHA formed native trimers. In contrast, the unmodified sHA was found to present epitopes recognized by a low-pH conformation specific monoclonal antibody. We found that mice primed and boosted with 3 µg of trimeric sHA in the absence of adjuvants had significantly higher IgG and HAI titers than mice that received the unmodified sHA. This correlated with an increased survival and reduced body weight loss following lethal challenge with mouse-adapted A/Aichi/2/68 virus. In addition, mice receiving a single vaccination of the trimeric sHA in the absence of adjuvants had improved survival and body weight loss compared to mice vaccinated with the unmodified sHA. Conclusions/Significance Our data indicate that the recombinant trimeric sHA presents native trimeric epitopes while the unmodified sHA presents epitopes not exposed in the native HA molecule. The epitopes presented in the unmodified sHA constitute a “silent face” which may skew the antibody response to epitopes not accessible in live virus at neutral pH. The results demonstrate that the trimeric sHA is a more effective influenza vaccine candidate and emphasize the importance of structure-based antigen design in improving recombinant HA vaccines.

• Publication year

  2010

• Venue

  PLoS ONE

• Publication date

  2010-09-01

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1371/journal.pone.0012466PMID 20824188PMCID 2931692
• 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.

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  2008cited by this paper
• An Influenza A Vaccine Based on Tetrameric Ectodomain of Matrix Protein 2*

  2008cited by this paper
• Safety and immunogenicity of a baculovirus-expressed hemagglutinin influenza vaccine: a randomized controlled trial.

  2007influential reference
• Influenza vaccine: the challenge of antigenic drift.

  2007cited by this paper
• Influenza viruses in animal wildlife populations.

  2007cited by this paper
• Changing perspective on immunization against influenza.

  2007influential reference
• Incorporation of High Levels of Chimeric Human Immunodeficiency Virus Envelope Glycoproteins into Virus-Like Particles

  2007cited by this paper
• Dose-related safety and immunogenicity of a trivalent baculovirus-expressed influenza-virus hemagglutinin vaccine in elderly adults.

  2006cited by this paper
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  2006cited by this paper
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  2006cited by this paper
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  2005cited by this paper
• Expression and purification of an influenza hemagglutinin—one step closer to a recombinant protein-based influenza vaccine

  2005cited by this paper
• Molecular and immunological characterization of soluble aggregated A/Victoria/3/75 (H3N2) influenza haemagglutinin expressed in insect cells

  2005cited by this paper
• Immunization against influenza A virus: comparison of conventional inactivated, live-attenuated and recombinant baculovirus produced purified hemagglutinin and neuraminidase vaccines in a murine model system.

  2005cited by this paper
• WHO Manual on Animal Influenza Diagnosis and Surveillance

  2002cited by this paper
• Safety and immunogenicity of a recombinant hemagglutinin vaccine for H5 influenza in humans.

  2001cited by this paper
• Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin.

  2000cited by this paper
• Regulation of antibody responses via antibodies, complement, and Fc receptors.

  2000cited by this paper
• Modifications That Stabilize Human Immunodeficiency Virus Envelope Glycoprotein Trimers in Solution

  2000cited by this paper
• Baculovirus-derived hemagglutinin vaccines protect against lethal influenza infections by avian H5 and H7 subtypes.

  1999cited by this paper
• Immunization with influenza A virus hemagglutinin and neuraminidase produced in recombinant baculovirus results in a balanced and broadened immune response superior to conventional vaccine.

  1999cited by this paper
• Effector CD4+ and CD8+ T‐cell mechanisms in the control of respiratory virus infections

  1997cited by this paper
• A switch between two-, three-, and four-stranded coiled coils in GCN4 leucine zipper mutants.

  1993cited by this paper
• Structural assignment of novel and immunodominant antigenic sites in the neutralizing antibody response of CBA/Ca mice to influenza hemagglutinin

  1991cited by this paper
• Structural basis of immune recognition of influenza virus hemagglutinin.

  1990cited by this paper
• Lymphokine control of in vivo immunoglobulin isotype selection.

  1990cited by this paper
• The IgG subclass responses to influenza virus haemagglutinin in the mouse: effect of route of inoculation.

  1989cited by this paper
• Evidence that the leucine zipper is a coiled coil.

  1989cited by this paper
• Studies on the structure of the influenza virus haemagglutinin at the pH of membrane fusion.

  1988cited by this paper
• The structure and function of the hemagglutinin membrane glycoprotein of influenza virus.

  1987cited by this paper
• Conformational changes in influenza virus haemagglutinin and its monomer detected by monoclonal antibodies.

  1985cited by this paper
• Changes in the conformation of influenza virus hemagglutinin at the pH optimum of virus-mediated membrane fusion.

  1982cited by this paper
• Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 Å resolution

  1981cited by this paper
• The role of serum haemagglutination-inhibiting antibody in protection against challenge infection with influenza A2 and B viruses

  1972cited by this paper
• Use of dimethyl suberimidate, a cross-linking reagent, in studying the subunit structure of oligomeric proteins.

  1970cited by this paper

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  2015cites this paper
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  2015cites this paper
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  2015cites this paper
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  2015cites this paper
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  2015cites this paper
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  2014cites this paper
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  2014cites this paper
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  2014cites this paper
• Influenza virus hemagglutinin as a vaccine antigen produced in bacteria.

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  2014cites this paper
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