Development of Zika viral pseudoinfectious virus as zika vaccine candidate Abstract Zika virus(ZIKV) is a mosquito-borne flavivirus that was first identified in Uganda in 1947. It then spread to Asia and Pacific. Recently, the explosive outbreaks in Americas and its implication in Guillan Barré (GBS) syndrome and the birth of microcephalic infants following maternal infection lead to WHO declared that ZIKV infection is a Public Health Emergency of International Concern. Currently, there is no specific treatment. Thus, developing a safe and effective vaccine for ZIKV infection is a high priority for public health. Here, we propose to develop a novel ZIKV vaccine candidate based on ZIKV pseudoinfectious virus (PIV) produced by incorporation of ZIKV subgenomic replicon into viral particle in stable packaging cells. The approach is based on our extensive experience of dengue virus (DENV) PIV investigation. In DENV PIV production system, the subgenomic replicon was constructed by deleting the capsid protein (C) gene from the DENV genome and optimizing the signal peptide sequence of pre- membrane protein (prM) to facilitate the formation of viral particles. Packaging cells were developed for inducible expression of a bi-protein Cpr, where the protein pr is the pr segment of viral protein prM that holds the protein C on the endoplasmic reticulum (ER). When the replicon was introduced into the packaging cells, protein C was released from the bi-protein Cpr by a replicon-encoded viral protease. Coordinate expression of viral structural proteins by the replicon and packaging cells led to the incorporation of the replicon into viral particle to produce DENV PIVs. Toward to develop a ZIKV PIV, we have constructed a full-length cDNA clone of ZIKV by chemical synthesis. Based upon these highly promising results, the Specific Aims of this Phase I STTR proposal are: 1) construction of subgenome for ZIKV 2) development of stable packaging cell lines based on FDA approved Vero cells, 3) rapid and efficient harvesting of ZIKV PIV from infected packaging lines, and 4) preliminary analysis of the safety and immunogenicity of the proposed vaccine in mice. Successful completion of this program will enable a Phase II research including development of a cost-effective GMP process for PIVs production at industrial scale and a pre-clinical animal study of immunity to ZIKV infection. Public Health Relevance Statement Pseudoinfectious virus (PIV) may induce immunity profile of live viral vaccine with the safety character of subunit vaccine. In the proposed project, the Zika virus (ZIKV) PIV vaccine will be produced in highly efficient Vero-based packaging cells. The immunogenicity of ZIKV PIV will be investigated in a novel mice model.
Project Terms: Aedes; Americas; Animals; immunogen; ATGN; Antigens; virus antigen; Viral Antigens; Asia; Parturition; Birth; Brazil; cDNA; Complementary DNA; cultured cell line; Strains Cell Lines; CellLine; Cell Line; Cell Body; Cells; chemical synthesis; breakbone fever; Dengue Fever; Dengue; Breakbone Fever Virus; Dengue Virus; Outbreaks; Disease Outbreaks; drug/agent; Pharmaceutic Preparations; Medication; Drugs; Pharmaceutical Preparations; Elements; Japanese B Viral Encephalitis; Japanese B Encephalitis; Japanese Encephalitis; Ergastoplasm; Endoplasmic Reticulum; Epidemic; Group B Arbovirus; Flaviviridae; Flavivirus; Foundations; French Polynesia; Genes; Genome; Goals; postinfectious polyneuritis; acute postinfectious polyneuropathy; acute idiopathic polyneuritis; Landry-Guillain-Barre Syndrome; Landry's paralysis; Guillaine-Barre Syndrome; Acute Inflammatory Polyradiculoneuropathy; Acute Inflammatory Polyneuropathy; Acute Inflammatory Demyelinating Polyradiculoneuropathy; Acute Infective Polyneuritis; Acute Autoimmune Neuropathy; AIDP; Guillain-Barre Syndrome; individuals with disabilities; disabled people; disabled individual; disabled; Persons with Disabilities; People with Disabilities; Handicapped; Disabled Population; Disabled Persons; Helper Viruses; Immunity; In Vitro; Industrialization; Infant; Infection; Lead; heavy metal lead; heavy metal Pb; Pb element; Lethal Dose 50; LD50; LD-50; Membrane Proteins; Surface Proteins; Membrane-Associated Proteins; Membrane Protein Gene; Microcephaly; microencephaly; micrencephaly; Micronesia; Culicidae; Mosquitoes; Mus; Murine; Mice Mammals; Mice; Pan American Health Organization; PAHO; Peptide Hydrolases; Proteolytic Enzymes; Proteinases; Proteases; Protease Gene; Peptidases; Esteroproteases; Production; Protein C; Proteins; Public Health; Replicon; Replication Unit; Research; Safety; Serotyping; Peptide Signal Sequences; protein signal sequence; Signal Sequences; Signal Peptide; Syndrome; Teratogens; Teratogenicity; Teratogenic; Uganda; United States; Vaccines; Vero Cells; virus genome; Viral Genome; virus protein; Viral Gene Proteins; Viral Gene Products; Viral Proteins; Viral Structural Proteins; Viral Vaccines; Virus Particle; Virion; virus-induced disease; virus infection; viral infection; Viral Diseases; Virus Diseases; General Viruses; Virus; West Nile; WNV; Egypt 101 virus; West Nile virus; Yellow Fever; Yellow fever virus; Titrations; base; genetic manipulation; Phase; Medical; Series; Infectious Agent; infectious organism; Attenuated; DNA Vaccines; Recombinant DNA Vaccines; Naked DNA Vaccines; programs; Investigation; Neuropathy; neuropathic; System; Location; Country; Viral; experience; particle; novel; Reporting; response; Subunit Vaccines; Formalin; Capsid Proteins; Viral Outer Coat Protein; Viral Coat Proteins; Coat Proteins; Length; Preventive; Harvest; International; Recombinants; in vivo; STTR; Small Business Technology Transfer Research; Process; developmental; Development; preclinical; pre-clinical; vector mosquito; immunogenicity; cost effective; cost-effective; pathogen; mouse model; murine model; FDA approved; vaccine candidate; public health emergency; public health priorities; co-infection; coinfection; Zika Virus; zikav; ZIKV; ZIKA; inducible gene expression; inducible expression; Structural Protein; Zika virus vaccine; Zika vaccine
