Antigenicity, stability, and reproducibility of Zika reporter virus particles for long-term applications
| Autor: | Edgar Davidson, J. Charles Whitbeck, M. Javad Aman, Graham Simmons, Lewis J. Stafford, Kathryn Kadash-Edmondson, Benjamin J. Doranz, Ariadna Grinyo-Escuer, Anu Thomas, Frederick W. Holtsberg, Grant C. Liao, Celine Cheng |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
RNA viruses Physiology RC955-962 Dengue virus medicine.disease_cause Pathology and Laboratory Medicine Antibodies Viral Biochemistry Neutralization Zika virus Genes Reporter Immune Physiology Cricetinae Arctic medicine. Tropical medicine Chlorocebus aethiops Medicine and Health Sciences Mass Screening Enzyme-Linked Immunoassays Immune System Proteins biology Zika Virus Infection Luciferase Assay Antibodies Monoclonal Enzymes Titer Infectious Diseases Bioassays and Physiological Analysis Medical Microbiology Viral Pathogens Viruses Pathogens Public aspects of medicine RA1-1270 Oxidoreductases Luciferase Research Article Antigenicity 030106 microbiology Immunology Research and Analysis Methods Microbiology Virus Antibodies Cell Line 03 medical and health sciences Plaque reduction neutralization test Neutralization Tests Virology medicine Animals Humans Antigens Immunoassays Microbial Pathogens Vero Cells Enzyme Assays Biology and life sciences Flaviviruses Public Health Environmental and Occupational Health Organisms Proteins Viral Vaccines Zika Virus Dengue Virus biology.organism_classification Antibodies Neutralizing Viral Replication High-Throughput Screening Assays 030104 developmental biology HEK293 Cells Viral replication Enzymology Immunologic Techniques Biochemical Analysis |
| Zdroj: | PLoS Neglected Tropical Diseases, Vol 14, Iss 11, p e0008730 (2020) PLoS Neglected Tropical Diseases |
| ISSN: | 1935-2735 1935-2727 |
| Popis: | The development of vaccines against flaviviruses, including Zika virus (ZIKV) and dengue virus (DENV), continues to be a major challenge, hindered by the lack of efficient and reliable methods for screening neutralizing activity of sera or antibodies. To address this need, we previously developed a plasmid-based, replication-incompetent DENV reporter virus particle (RVP) production system as an efficient and safe alternative to the Plaque Reduction Neutralization Test (PRNT). As part of the response to the 2015–2016 ZIKV outbreak, we developed pseudo-infectious ZIKV RVPs by modifying our DENV RVP system. The use of ZIKV RVPs as critical reagents in human clinical trials requires their further validation using stability and reproducibility metrics for large-scale applications. In the current study, we validated ZIKV RVPs using infectivity, neutralization, and enhancement assays with monoclonal antibodies (MAbs) and human ZIKV-positive patient serum. ZIKV RVPs are antigenically equivalent to live virus based on binding ELISA and neutralization results and are nonreplicating based on the results of live virus replication assays. We demonstrate reproducible neutralization titer data (NT50 values) across different RVP production lots, volumes, time frames, and laboratories. We also show RVP stability across experimentally relevant time intervals and temperatures. Our results demonstrate that ZIKV RVPs provide a safe, high-throughput, and reproducible reagent for large-scale, long-term studies of neutralizing antibodies and sera, which can facilitate large-scale screening and epidemiological studies to help expedite ZIKV vaccine development. Author summary ZIKV is a mosquito-borne virus that can cause severe birth defects and other disorders. Large outbreaks of ZIKV occurred in 2015 and 2016 and there are still no drugs or vaccines available to protect against ZIKV infection. Vaccine development has been hindered by the lack of safe and efficient methods to screen ZIKV neutralizing properties of patient sera or antibodies, especially in the context of large clinical trials. To address this unmet need, we developed and validated the use of ZIKV reporter virus particles (RVPs), a safe, high-throughput, and quantitative alternative to using live virus for neutralization studies. We show that ZIKV RVPs are stable, show lot-to-lot consistency, and provide reproducible neutralization data that is suitable for the large-scale studies needed for development of a ZIKV vaccine, epidemiologic surveillance, and high-throughput screening. |
| Databáze: | OpenAIRE |
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