Nucleic acids encoding Zika virus-like particles and their use in Zika virus vaccines and diagnostic assays
Inventors
Chang, Gwong-Jen J. • Davis, Brent S.
Assignees
US Department of Health and Human Services
Publication Number
US-11578103-B2
Publication Date
2023-02-14
Expiration Date
2037-06-09
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Abstract
Transcriptional units encoding Zika virus (ZIKV) premembrane (prM) and envelope (E) proteins, which upon translation form Zika virus-like particles (VLPs), are described. Use of the transcriptional units and VLPs in three different ZIKV vaccine platforms is described. Immunoassay-based detection methods using ZIKV VLPs are described for the diagnosis of ZIKV infection.
Core Innovation
The invention provides transcriptional units encoding Zika virus (ZIKV) premembrane (prM) and envelope (E) proteins, which upon translation form Zika virus-like particles (VLPs). These transcriptional units and VLPs are utilized in various ZIKV vaccine platforms including plasmid DNA vaccines, recombinant adenovirus vectors, and as isolated VLP immunogenic compositions. Additionally, the invention discloses immunoassay-based detection methods using ZIKV VLPs for diagnosing ZIKV infection.
The disclosed nucleic acid molecules include sequences for a modified Japanese encephalitis virus (JEV) signal sequence and a ZIKV prME coding sequence, operably linked to promoters such as the cytomegalovirus (CMV) promoter, and include translation initiation and transcription termination sequences. The constructs are designed for optimized expression in human cells, and some include amino acid substitutions in the E protein to reduce flavivirus cross-reactive immune responses. The recombinant adenoviruses express ZIKV VLPs upon transduction of host cells, serving as vaccine candidates that elicit protective immunity in animal models.
The background problem addressed is the global health threat posed by the emergence and rapid spread of ZIKV, causing congenital birth abnormalities, Guillain-Barré syndrome, encephalitis, and myelopathy. Diagnostic challenges arise due to the short viremic phase of humans, limiting nucleic acid-based detection. There is a need for improved serodiagnostic assays and effective vaccines to comprehensively mitigate ZIKV's impact. The invention aims to fulfill these unmet needs by providing vaccine components producing immunogenic VLPs and improved detection methods based on these VLPs.
Claims Coverage
The patent includes 20 claims covering two independent claims directed to a vector comprising a nucleic acid molecule and a recombinant adenovirus, each comprising a transcriptional unit encoding a modified JEV signal sequence and a ZIKV prME coding sequence with specific amino acid substitutions.
Vector comprising a transcriptional unit with modified JEV signal sequence and engineered ZIKV prME coding sequence
The vector includes a transcriptional unit with a sequence encoding a modified Japanese encephalitis virus (JEV) signal sequence (SEQ ID NO: 4) and a Zika virus (ZIKV) premembrane (prM) and envelope (E) protein coding sequence. The E protein comprises a lysine substitution at position 106 and an aspartic acid substitution at position 107 based on ZIKV strain MR766.
Transcriptional unit with regulatory sequences enhancing expression
The transcriptional unit further comprises a promoter operably linked to the prME coding sequence, specifically the cytomegalovirus (CMV) E1A promoter, a transcription termination sequence including bovine growth hormone (BGH) transcription termination sequence, and a translation initiation sequence comprising the Kozak consensus sequence GCCGCCGCCATGG (SEQ ID NO: 8).
Use of specific ZIKV strains and codon-optimization
The ZIKV strain encoded by the transcriptional unit is selected from MR-766, SPH2015, P6-740, and FSS 13025. The prME coding sequence is codon-optimized for expression in human cells and is at least 95% identical to defined nucleotide sequences (nucleotides 1186-3204 of SEQ ID NO: 20 or nucleotides 1186-3210 of SEQ ID NO: 22).
Recombinant adenovirus expressing ZIKV VLPs with engineered prME
Recombinant adenoviruses comprise the nucleic acid molecule encoding the modified JEV signal sequence and ZIKV prME coding sequence with lysine and aspartic acid substitutions at E positions 106 and 107, enabling expression of ZIKV virus-like particles upon transduction of host cells.
The claims cover vectors and recombinant adenoviruses comprising transcriptional units encoding ZIKV prM and E proteins with specific modifications to the E protein and regulatory elements to optimize expression and immunogenicity, forming the basis for vaccine development against Zika virus.
Stated Advantages
ZIKV prME transcriptional units and VLPs provide a versatile platform for vaccine development across different vaccine modalities, including DNA vaccines and recombinant adenovirus vectors.
The modified JEV signal sequence and codon optimization improve protein expression, translocation, and VLP secretion, enhancing immunogenicity.
Amino acid substitutions at E protein positions 106 and 107 reduce flavivirus cross-reactive immune responses, potentially increasing vaccine safety by diminishing antibody-dependent enhancement.
The recombinant adenovirus vaccine induces protective immunity in animal models with a single dose and provides protection against lethal ZIKV challenge.
Use of ZIKV VLPs enables sensitive and specific immunoassay detection methods for identifying ZIKV-specific antibodies, overcoming challenges related to the short viremic phase.
Documented Applications
Use of transcriptional units encoding ZIKV prM and E proteins to form virus-like particles (VLPs) for producing ZIKV vaccines in plasmid DNA, adenovirus vector, and isolated VLP formats.
Use of recombinant adenovirus vectors expressing ZIKV VLPs as a vaccine platform capable of eliciting protective immunity in animal models.
Methods of eliciting immune responses and immunizing subjects, specifically humans, against ZIKV by administering nucleic acid molecules, vectors, recombinant adenoviruses, or VLP compositions.
Use of ZIKV VLPs in serodiagnostic assays, such as IgM and IgG antibody capture ELISAs, microsphere immunoassays (MIA), and indirect ELISAs, for detecting ZIKV-specific antibodies in biological samples including serum, blood, and plasma.
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