Broad-spectrum antiviral brincidofovir inhibits Epstein-Barr virus and related gammaherpesvirus in human and nonhuman primate cells
Abaigeal Donaldson, Madeleine R. Druker, Maria Chiara Monaco, Emily H. Stack, Paige Zimmerman, Amanda Lee, Izabela Bialuk, William Frazier, Irene Cortese, Heather Narver, Masatoshi Hazama, Fuminori Yoshida, Xiaofan Li, Laurie T. Krug, Stacey L. Piotrowski, Steven Jacobson
Abaigeal Donaldson, Madeleine R. Druker, Maria Chiara Monaco, Emily H. Stack, Paige Zimmerman, Amanda Lee, Izabela Bialuk, William Frazier, Irene Cortese, Heather Narver, Masatoshi Hazama, Fuminori Yoshida, Xiaofan Li, Laurie T. Krug, Stacey L. Piotrowski, Steven Jacobson
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Research Article Neuroscience Virology

Broad-spectrum antiviral brincidofovir inhibits Epstein-Barr virus and related gammaherpesvirus in human and nonhuman primate cells

Abstract

Epstein-Barr virus (EBV) is of growing interest for its potential role in neurodegenerative diseases such as multiple sclerosis (MS) and its possible utility as a therapeutic target in herpesvirus-associated chronic diseases. The effects of brincidofovir (BCV) on EBV reactivation were evaluated in vitro using EBV-infected spontaneous lymphoblastoid cell lines (SLCLs) and peripheral blood mononuclear cells (PBMCs) derived from patients with MS and healthy controls. In addition, a B lymphoblastoid cell line and PBMCs from common marmosets (Callithrix jacchus) naturally infected with an EBV-related gammaherpesvirus (Callitrichine herpesvirus 3, CalHV-3) were used to measure BCV efficacy in a nonhuman primate model. BCV significantly inhibited gammaherpesvirus reactivation, with decreased lytic and latent viral transcript expression. These results suggest that BCV may be a useful antiviral for inhibiting EBV activity in patients with MS. Additionally, this work further validates the utility of CalHV-3 in marmosets as a translational model for the investigation of successful EBV-targeting therapeutics.

Authors

Abaigeal Donaldson, Madeleine R. Druker, Maria Chiara Monaco, Emily H. Stack, Paige Zimmerman, Amanda Lee, Izabela Bialuk, William Frazier, Irene Cortese, Heather Narver, Masatoshi Hazama, Fuminori Yoshida, Xiaofan Li, Laurie T. Krug, Stacey L. Piotrowski, Steven Jacobson

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Figure 5

CalHV-3–infected marmoset cells treated with brincidofovir show decreases in CalHV-3 DNA and mRNA following viral reactivation assay.

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CalHV-3–infected marmoset cells treated with brincidofovir show decrease...
(A–C) Quantification of Callitrichine herpesvirus 3 (CalHV-3) DNA and mRNA expression in CalHV-3 infected cell line, CJ0149, with (dark blue bars, right) or without (light blue bar, left) NaB+TPA to induce viral reactivation. (A) CalHV-3 DNA expression; (B) ORF39 mRNA expression; (C) ORF45 mRNA expression. (D–F) Representative ddPCR plots of ORF39 mRNA expression in marmoset #3 PBMCs cultured (D) without NaB+TPA and no BCV, (E) with NaB+TPA and no BCV, and (F) with NaB+TPA and 250nM BCV. Y axis represents FAM-MGBNFQ fluorescent amplitude, which corresponds to ORF39 mRNA detection (blue dots + orange dots, upper quadrants). X axis represents the VIC-MGBNFQ fluorescent amplitude, which corresponds to the CJ-TBP housekeeping gene (green dots, bottom right quadrant). Lower left quadrant represents droplets negative for both ORF39 and CJ-TBP (gray dots). (G–I) Mean detection ± SEM of CalHV-3 gene expression in marmoset PBMCs (n = 4, circles) cultured with BCV and with (dark blue bars, right) or without (light blue bar, left) NaB+TPA to induce viral reactivation. (G) CalHV-3 DNA expression; (H) ORF39 mRNA expression; (I) ORF45 mRNA expression. Data analyzed using (G) Kruskal-Wallis test with Dunn’s comparisons, (H) a Welch’s ANOVA, and (I) 1-way ANOVA with Tukey’s multiple comparisons, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. Control, cells treated with only media; BCV, brincidofovir.