Abstract
Yellow fever virus (YFV), a member of the Flaviviridae family, is an arthropod-borne virus that can cause severe disease in humans with a lethality rate of up to 60%. Since 2017, increases in YFV activity in areas of South America and Africa have been described. Although a vaccine is available, named strain 17D (Theiler and Smith, 1937), it is contraindicated for use in the elderly, expectant mothers, immunocompromised people, among others. To this day there is no antiviral treatment against YFV to reduce the severity of viral infection. Here, we used a circular polymerase extension reaction (CPER)-based reverse genetics approach to generate a full-length reporter virus (YFVhb) by introducing a small HiBit tag in the NS1 protein. The reporter virus replicates at a similar rate to the parental YFV in HuH-7 cells. Using YFVhb, we designed a high throughput antiviral screening luciferase-based assay to identify inhibitors that target any step of the viral replication cycle. We validated our assay by using a range of inhibitors including drugs, immune sera and neutralizing single chain variable fragments (scFv). In light of the recent upsurge in YFV and a potential spread of the virus, this assay is a further tool in the development of antiviral therapy against YFV.
Highlights
Yellow fever virus (YFV), a Flaviviridae family member, is an arthropod borne virus mainly transmitted to vertebrate hosts by mosquitoes of the Aedes and Haemagogus species (Huang, 1986; Monath, 1994; Mutebi and Barrett, 2002; Vasconcelos et al, 2003)
The circular polymerase extension reaction (CPER) approach has been previously successfully used for ZIKV rescue (Setoh et al, 2017) and tagging with HiBiT has been established for other flaviviruses (Tamura et al, 2017, 2019)
Reporter viruses containing a full reporter protein in the capsid region are often unstable, attenuated, or display strong differences in viral growth compared to parental virus (Gadea et al, 2016; Schoggins et al, 2012; Vandergaast et al, 2014; Zou et al, 2011)
Summary
Yellow fever virus (YFV), a Flaviviridae family member, is an arthropod borne virus mainly transmitted to vertebrate hosts by mosquitoes of the Aedes and Haemagogus species (Huang, 1986; Monath, 1994; Mutebi and Barrett, 2002; Vasconcelos et al, 2003). A vaccine against YFV exists, limitations with the vaccine formulation, severe adverse effects, and shortage of supply have boosted the interest in effective yet safer vaccine strategies (Julander et al, 2018; Monath et al, 2010; Pato et al, 2019; Tottey et al, 2018). There is no therapeutic treatment to reduce the severity of YFV infections and patients rely on palliative care. The lethality of YFV has been reported to be as high as 60% (Monath, 2008; Theiler and Smith, 1937). Rapid and sensitive assays that are amenable to high-throughput
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