Zika virus vaccines
Inventors
Graham, Barney S. • PIERSON, Theodore C. • DOWD, Kimberly A. • Mascola, John R. • Kong, Wing-pui • Ko, Sung-Youl • YANG, Eun Sung • Shi, Wei • Wang, Lingshu • DEMASO, Christina R. • PELC, Rebecca S. • CREANGA, Adrian • Ledgerwood, Julie • CASTILHO, Leda R.
Assignees
US Department of Health and Human Services
Publication Number
US-10898566-B2
Publication Date
2021-01-26
Expiration Date
2037-07-28
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Abstract
The present invention relates to a vaccine for Zika virus, the vaccine comprising Zika virus membrane and envelope proteins. More specifically, the vaccine comprises nucleic acid molecules encoding modified Zika virus membrane and/or envelope proteins. When introduced into a cell, the encoded proteins are produced, which results in the production of a virus-like particle capable of eliciting an immune response against Zika virus.
Core Innovation
The invention provides a vaccine for Zika virus comprising nucleic acid molecules encoding modified Zika virus membrane and envelope proteins that, when expressed in a cell, produce virus-like particles (VLPs) capable of eliciting an immune response against Zika virus. More specifically, the vaccine includes a nucleic acid molecule encoding a polyprotein comprising at least a portion of a Zika virus premembrane (prM) protein joined to at least a portion of a Zika virus envelope (E) protein, where the prM protein portion includes a signal sequence heterologous to Zika virus. Expression of this polyprotein in cells leads to production of VLPs that mimic native virus structure and induce protective immunity.
The problem addressed is that prior expression of Zika virus structural proteins from cDNA results in inefficient cleavage at the amino terminus of prM, leading to deficient secretion of prM-E heterodimers and a lack of immunogenicity. Furthermore, existing flavivirus vaccines face challenges due to viral genetic diversity and limitations of eliciting broadly neutralizing antibodies. There is a need for a safe and effective vaccine for flaviviruses, including Zika virus, which elicits robust and protective neutralizing antibody responses.
The invention solves this problem by introducing a heterologous signal sequence to improve prM cleavage and secretion, and by modifying the envelope protein, including substitution of the stem and transmembrane regions with corresponding regions from related flaviviruses such as Japanese Encephalitis Virus. These modifications enhance production and immunogenicity of VLPs. The vaccine induces neutralizing antibody titers correlated with protection, demonstrated in animal models and clinical trials, thereby providing a new approach for effective Zika virus immunization.
Claims Coverage
The patent includes 20 inventive features primarily concerning nucleic acid molecules encoding modified Zika virus polyproteins, methods of producing virus-like particles, and methods of eliciting immune responses.
Nucleic acid molecule encoding a chimeric polyprotein
A nucleic acid molecule encoding a polyprotein comprising at least a portion of a Zika virus premembrane (prM) protein joined to a portion of a Zika virus envelope (E) protein, wherein the prM portion includes a signal sequence heterologous to Zika virus and the envelope protein stem and/or transmembrane domains are from a flavivirus other than Zika virus.
Heterologous signal sequence source
The heterologous signal sequence is from a flavivirus prM protein, human CD5 protein, mouse IL-2 protein, or bovine prolactin, with flaviviruses selected from Japanese encephalitis virus, yellow fever virus, Dengue virus, and West Nile Virus.
Replacement of envelope protein stem/transmembrane regions
The stem and/or transmembrane domains of the envelope protein are from Japanese Encephalitis Virus, specifically comprising sequences at least 85% identical to designated SEQ ID NOs related to JEV stem/transmembrane sequences.
Signal sequence comprising the Japanese Encephalitis Virus envelope protein signal sequence
The nucleic acid molecule's signal sequence comprises the Japanese Encephalitis Virus envelope protein signal sequence or sequences highly identical thereto.
Variants with specific sequence identities and mutations
The Zika virus prM protein comprises amino acid sequences at least 80%-97% identical to selected SEQ ID NOs (29-239) and includes at least one mutation from this group; similarly, the polyprotein can be at least 80%-97% identical to SEQ ID NO:110 or SEQ ID NO:114.
Methods of producing Zika virus-like particles
Introducing the nucleic acid molecule into a cell to express the polyprotein and produce virus-like particles capable of eliciting immune responses.
Methods of eliciting immune responses
Administering the nucleic acid molecule to an individual to induce an immune response against Zika virus.
Signal sequence with specific sequence identity
The signal sequence comprises amino acid sequences at least 90% identical to SEQ ID NOs: 18, 20, 22, 24, or 26.
Portions of prM protein with specified identity
The prM protein portion comprises at least 50 contiguous amino acids with at least 80% identity to SEQ ID NO:2 or comprises an amino acid sequence at least 80% identical to SEQ ID NO:2.
Portions of envelope protein with specified identity
The envelope protein portion comprises at least 50 contiguous amino acids with at least 80% identity to SEQ ID NO:4 or comprises an amino acid sequence at least 80% identical to SEQ ID NO:4.
Stem domain sequence identity
The stem domain in the envelope protein comprises an amino acid sequence at least 85% identical to SEQ ID NO:8.
Transmembrane domain sequence identity
The transmembrane domain in the envelope protein comprises an amino acid sequence at least 85% identical to SEQ ID NO:12.
Region corresponding to stem/transmembrane domains
The region corresponding to stem and transmembrane domains comprises an amino acid sequence at least 85% identical to SEQ ID NO:16.
Kit including the nucleic acid molecule
A kit comprising the nucleic acid molecule encoding the modified polyprotein as described.
Polyprotein mutations at specific residues
The polyprotein comprises at least one mutation in the prM protein at a position corresponding to amino acid H7 of SEQ ID NO:2, or in the envelope protein at amino acid positions selected from a specified group of residues in SEQ ID NO:4, including R2, G5, N8, S16, and others.
Polyprotein sequence identity to modified sequences
The polyprotein comprises an amino acid sequence at least 90%-99% identical to sequences selected from SEQ ID NO:29-239.
The claims cover nucleic acid molecules encoding chimeric and modified Zika virus polyproteins with heterologous signal sequences and envelope protein domains from other flaviviruses, methods for producing VLPs and eliciting immunity, sequence variants with defined identities and mutations enhancing immunogenicity, and kits for these molecules.
Stated Advantages
Provides a safe and effective vaccine against Zika virus capable of eliciting robust neutralizing antibody responses.
Improved production and secretion of prM-E heterodimers leading to enhanced immunogenicity over prior constructs.
Ability to elicit broadly neutralizing antibodies against different Zika virus strains including Asian and African lineages.
Use of virus-like particles with native-like three-dimensional structures that improve immune response elicitation.
Potent immunogenicity demonstrated in animal models and human clinical trials.
Potential for rapid vaccine development and deployment due to use of nucleic acid and VLP platforms.
Documented Applications
Vaccines for immunizing individuals against Zika virus infection.
Methods for eliciting immune responses against Zika virus via administration of nucleic acid molecules, proteins, or VLPs.
Production of virus-like particles and reporter virus particles for use as vaccines or diagnostic reagents.
Use of VLPs and RVPs for detecting anti-Zika virus antibodies in samples from individuals.
Methods to identify individuals exposed to Zika virus by serological testing using VLPs or RVPs.
Measuring the immune response of individuals to Zika virus vaccines using VLP- or RVP-based assays.
Clinical trials assessing the immunogenicity and safety of DNA vaccines encoding these modified Zika virus proteins.
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