title: Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons.
abstract: Zika Virus (ZIKV) is an emerging pathogen that is linked to fetal developmental abnormalities such as microcephaly, eye defects, and impaired growth. ZIKV is an RNA virus of the Flaviviridae family. ZIKV is mainly transmitted by mosquitoes, but can also be spread by maternal to fetal vertical transmission as well as sexual contact. To date, there are no reliable treatment or vaccine options available to protect those infected by the virus. The development of a reproducible, effective Zika virus infectious cell culture system is critical for studying the molecular mechanisms of ZIKV replication as well as drug and vaccine development. In this regard, a protocol describing a mammalian cell-based in vitro Zika virus culture system for viral production and growth analysis is reported here. Details on the formation of plaques by Zika virus on a cell monolayer and plaque assay for measuring viral titer are presented. Viral genome replication kinetics and double-stranded RNA genome replicatory intermediates are determined. This culture platform was utilized to screen against a library of a small set of cytokines resulting in the identification of interferon-α (IFN-α), IFN-β and IFN-γ as potent inhibitors of Zika viral growth. In summary, an in vitro infectious Zika viral culture system and various virological assays are demonstrated in this study, which has the potential to greatly benefit the research community in elucidating further the mechanisms of viral pathogenesis and the evolution of viral virulence. Antiviral IFN-alpha can further be evaluated as a prophylactic, post-exposure prophylactic, and treatment option for Zika virus infections in high-risk populations, including infected pregnant women.
authors: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center.
authors: Deisy
authors: D
authors: Contreras
authors: Board of Governors Regenerative Medicine Institute, Cedars-Sinai Medical Center; Department of Surgery, Cedars-Sinai Medical Center; Department of Surgery, David Geffen School of Medicine, University of California, Los Angeles; arumugaswami@cshs.org.
authors: Vaithilingaraja
authors: V
authors: Arumugaswami
mesh_terms: D000818:Animals; D002460:Cell Line; D005260:Female; D016679:Genome, Viral; D006801:Humans; D018445:Infectious Disease Transmission, Vertical; D007372:Interferons / Q000494:pharmacology*; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D019562:Viral Load; D010948:Viral Plaque Assay; D014779:Virus Replication; D000071244:Zika Virus; D000071243:Zika Virus Infection / Q000517:prevention & control* / Q000821:virology
chemical_list: D012330:RNA, Double-Stranded; D012367:RNA, Viral; D007372:Interferons