Self-assembling influenza nanoparticle vaccines elicit broadly neutralizing H1N1 antibodies
Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA.Nature (Impact Factor: 41.46). 05/2013; 499(7456). DOI: 10.1038/nature12202
Influenza viruses pose a significant threat to the public and are a burden on global health systems. Each year, influenza vaccines must be rapidly produced to match circulating viruses, a process constrained by dated technology and vulnerable to unexpected strains emerging from humans and animal reservoirs. Here we use knowledge of protein structure to design self-assembling nanoparticles that elicit broader and more potent immunity than traditional influenza vaccines. The viral haemagglutinin was genetically fused to ferritin, a protein that naturally forms nanoparticles composed of 24 identical polypeptides. Haemagglutinin was inserted at the interface of adjacent subunits so that it spontaneously assembled and generated eight trimeric viral spikes on its surface. Immunization with this influenza nanoparticle vaccine elicited haemagglutination inhibition antibody titres more than tenfold higher than those from the licensed inactivated vaccine. Furthermore, it elicited neutralizing antibodies to two highly conserved vulnerable haemagglutinin structures that are targets of universal vaccines: the stem and the receptor binding site on the head. Antibodies elicited by a 1999 haemagglutinin-nanoparticle vaccine neutralized H1N1 viruses from 1934 to 2007 and protected ferrets from an unmatched 2007 H1N1 virus challenge. This structure-based, self-assembling synthetic nanoparticle vaccine improves the potency and breadth of influenza virus immunity, and it provides a foundation for building broader vaccine protection against emerging influenza viruses and other pathogens.
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- "Ferritins have been engineered for multiple purposes (Huard et al., 2013; Jä a ¨ skelä inen et al., 2007; Li et al., 2006; Meldrum et al., 1992). Among these innovations, we have recently developed a platform to mount a viral trimeric glycoprotein at ferritin's 3-fold axes, providing a basis for structure-based nanoparticulate immunogens (Kanekiyo et al., 2013). Comparison of ferritin structures revealed that certain ferritins, including human light chain (Wang et al., 2006) (PDB: 2FFX) and bullfrog lower subunit (Trikha et al., 1995) (PDB: 1RCC), contained an NH 2 -terminal extension that was not present in ferritin from Helicobacter pylori (Cho et al., 2009) (PDB: 3EGM). "
ABSTRACT: Epstein-Barr virus (EBV) represents a major global health problem. Though it is associated with infectious mononucleosis and ∼200,000 cancers annually worldwide, a vaccine is not available. The major target of immunity is EBV glycoprotein 350/220 (gp350) that mediates attachment to B cells through complement receptor 2 (CR2/CD21). Here, we created self-assembling nanoparticles that displayed different domains of gp350 in a symmetric array. By focusing presentation of the CR2-binding domain on nanoparticles, potent neutralizing antibodies were elicited in mice and non-human primates. The structurally designed nanoparticle vaccine increased neutralization 10- to 100-fold compared to soluble gp350 by targeting a functionally conserved site of vulnerability, improving vaccine-induced protection in a mouse model. This rational approach to EBV vaccine design elicited potent neutralizing antibody responses by arrayed presentation of a conserved viral entry domain, a strategy that can be applied to other viruses. Copyright © 2015 Elsevier Inc. All rights reserved. Full text can be found at http://authors.elsevier.com/a/1RcBdL7PXI6a- Valid until October 16, 2015
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- "The highly symmetric and self-assembling ferritin nanocage presents an attractive target for vaccine development. In an elegant demonstration of the use of ferritin for vaccine development , Kanekiyo and colleagues fused the influenza virus haemagglutinin (HA) to the surface of Helicobacter pylori ferritin . Fusions of entire protein domains to the surface of ferritin are complicated by the requirement to match the subunit organisation of the protein to be displayed on its surface with the inherent symmetry of the ferritin cage itself. "
ABSTRACT: Ferritin family proteins are found in all kingdoms of life and act to store iron within a protein cage and to protect the cell from oxidative damage caused by the Fenton reaction. The structural and biochemical features of the ferritins have been widely exploited in bionanotechnology applications: from the production of metal nanoparticles; as templates for semi-conductor production; and as scaffolds for vaccine design and drug delivery. In this review we first discuss the structural properties of the main ferritin family proteins, and describe how their organisation specifies their functions. Second, we describe materials science applications of ferritins that rely on their ability to sequester metal within their cavities. Finally, we explore the use of ferritin as a container for drug delivery and as a scaffold for the production of vaccines. Copyright © 2015. Published by Elsevier B.V.
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- "In this study, we present the methods of construction, expression and purification of the truncated HA protein from H5N1 Malaysia strain in bacterial expression system, E. coli. HA, a major " antigenic change " envelope protein, is the primary target for neutralizing antibodies and therefore becomes the main focus of influenza vaccines development (Wei et al., 2012; Kanekiyo et al., 2013; Dilillo et al., 2014). Recently, Herfst et al. (2012) shows that only four amino acid substitutions in the hemagglutinin, and one in the polymerase basic 2, are capable to transform H5N1 virus be airborne transmitted between ferrets. "
ABSTRACT: Malaysia first reported H5N1 poultry case in 2004 and subsequently outbreak in poultry population in 2007. Here, a recombinant gene encoding of peptide epitopes, consisting fragments of HA1, HA2 and a polybasic cleavage site of H5N1 strain Malaysia, was amplified and cloned into pET-47b(+) bacterial expression vector. DNA sequencing and alignment analysis confirmed that the gene had no alteration and in-frame to the vector. Then, His-tagged truncated HA protein was expressed in Escherichia coli BL21 (DE3) under 1 mM IPTG induction. The protein expression was optimized under a time-course induction study and further purified using Ni-NTA agarose under reducing condition. Migration size of protein was detected at 15 kDa by Western blot using anti-His tag monoclonal antibody and demonstrated no discrepancy compared to its calculated molecular weight.
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