Response to reviewer 2

Wispelaere, Lien De

doi:10.5194/bg-2016-354-ac2

Cited by 1 publication

(5 citation statements)

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“…1A) in which single-cycle viral yield was quantified as a readout of productive viral entry. Analogous virological assays were used to demonstrate 3-110-22's activity against WNV, JEV, and ZIKV 22 . To ensure that the antiviral effects we observed is due to inhibition of E's function in viral entry and not due to other onor off-target effects, we limit inhibitor treatment to pre-incubation with the viral inoculum and the first hour of cellular infection, then wash the cells to remove extracellular virus and inhibitor prior to the addition of fresh medium.…”

Section: Resultsmentioning

confidence: 99%

“…Vero cells (ATCC, RRID:CVCL_0059), baby hamster kidney-derived BHK-21 cells (laboratory of Eva Harris, University of California, Berkeley), C6/36 cells derived from Aedes albopictus (Diptera: Culicidae) embryonic tissue (ATCC, RRID:CVCL_Z230), Spodoptera frugiperda cells (Sf9) (laboratory of Stephen C. Harrison, Cat # B825-01 Invitrogen/Thermo Fisher Scientific), and Trichoplusia ni cells (High Five ™ , laboratory of Stephen C. Harrison, Invitrogen/Thermo Fisher Scientific) were maintained as previously described [21][22] .…”

Section: Cell Linesmentioning

confidence: 99%

“…All work with infectious virus was performed in a biosafety level 2 (BSL2) laboratory using additional safety practices as approved by the Harvard Committee on Microbiological Safety. Dengue virus serotype 2 strain New Guinea C (DENV2 NGC; Lee Gehrke, MIT), dengue virus serotype 2 strain S221 (DENV2 S221; Sujan Shresta, La Jolla Institute for Allergy and Immunology), and Zika virus (ZIKV) strain PF-251013-18 (Didier Musso, Institut Louis Malardé, Tahiti Island, French Polynesia) were propagated in C6/36 cells as previously described [21][22] . The Japanese encephalitis virus (JEV) vaccine strain SA14-14-2 was kindly provided by Gregory Gromowski (Walter Reed Army Institute of Research).…”

Section: Virusesmentioning

confidence: 99%

“…DENV2 and JEV titers were quantified by plaque-formation assay (PFA) on BHK-21 cells, and ZIKV titers were quantified by PFA on Vero cells following published procedures [21][22] . Incubation times for plaque formation at 37 °C were 2.5 days (ZIKV), 3 days (JEV), or 4.5 days (DENV2).…”

Section: Plaque-forming Assay (Pfa) and Focus-forming Assaymentioning

confidence: 99%

“…The soluble DENV2 prefusion E dimer (sE 2 ) bearing a 6His-N-terminal purification tag and AviTag TdeM4 C-terminal tag as previously described [21][22] . Equilibrium dissociation constant for binding of inhibitor to sE 2 was measured by biolayer interferometry using an Octet RED384 system (FortéBio) using superstreptavidin (SSA) biosensors.…”

Section: Expression Purification and Biotinylation Of Recombinant Smentioning

confidence: 99%

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Small Molecules Targeting the Flavivirus E Protein with Broad-Spectrum Activity and Antiviral Efficacy in Vivo

Jang

Hsia

et al. 2019

ACS Infect. Dis.

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Vaccines and antivirals to combat dengue, Zika, and other flavivirus pathogens present a major, unmet medical need. Vaccine development has been severely challenged by the antigenic diversity of these viruses and the propensity of non-neutralizing, cross-reactive antibodies to facilitate cellular infection and increase disease severity. As an alternative, direct-acting antivirals targeting the flavivirus envelope protein, E, have the potential to act via an analogous mode of action without the risk of antibody-dependent enhancement of infection and disease. We previously discovered that structurally diverse small molecule inhibitors of the dengue virus E protein exhibit varying levels of antiviral activity against other flaviviruses in cell culture. Here we demonstrate that the broad-spectrum activity of several cyanohydrazones against dengue, Zika, and Japanese encephalitis viruses is due to specific inhibition of E-mediated membrane fusion during viral entry and provide proof of concept for pharmacological inhibition of E as an antiviral strategy in vivo.

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Section: Resultsmentioning

confidence: 99%

Section: Cell Linesmentioning

confidence: 99%

Section: Virusesmentioning

confidence: 99%

Section: Plaque-forming Assay (Pfa) and Focus-forming Assaymentioning

confidence: 99%

Section: Expression Purification and Biotinylation Of Recombinant Smentioning

confidence: 99%

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