Honokiol and Alpha-Mangostin Inhibit Mayaro Virus Replication through Different Mechanisms

Abstract

Mayaro virus (MAYV) is an emerging arbovirus with an increasing circulation across the Americas. In the present study, we evaluated the potential antiviral activity of the following natural compounds against MAYV and other arboviruses: Sanguinarine, (R)-Shikonin, Fisetin, Honokiol, Tanshinone IIA, and α-Mangostin. Sanguinarine and Shikonin showed significant cytotoxicity, whereas Fisetin, Honokiol, Tanshinone IIA, and α-Mangostin were well tolerated in all the cell lines tested. Honokiol and α-Mangostin treatment protected Vero-E6 cells against MAYV-induced damage and resulted in a dose-dependent reduction in viral progeny yields for each of the MAYV strains and human cell lines assessed. These compounds also reduced MAYV viral RNA replication in HeLa cells. In addition, Honokiol and α-Mangostin disrupted MAYV infection at different stages of the virus life cycle. Moreover, Honokiol and α-Mangostin decreased Una, Chikungunya, and Zika viral titers and downmodulated the expression of E1 and nsP1 viral proteins from MAYV, Una, and Chikungunya. Finally, in Honokiol- and α-Mangostin-treated HeLa cells, we observed an upregulation in the expression of type I interferon and specific interferon-stimulated genes, including IFNα, IFNβ, MxA, ISG15, OAS2, MDA-5, TNFα, and IL-1β, which may promote an antiviral cellular state. Our results indicate that Honokiol and α-Mangostin present potential broad-spectrum activity against different arboviruses through different mechanisms.

Differential susceptibility of human microglia HMC3 cells and brain microvascular endothelial HBEC-5i cells to Mayaro and Una virus infection

“…The Mayaro (MAYV, AVR 0565, Peru; MAYV, Guyane, French Gianna; and MAYV, TRVL 4675, Trinidad and Tobago) and Una (UNAV, BT 1495-3, Panama; UNAV, 788382, Trinidad and Tobago; and UNAV, CoAr 2518, Colombia) strains used in this work were obtained from the World Reference Center for Emerging Viruses and Arboviruses (WRCEVA) at University of Texas Medical Branch (UTMB, USA) and kindly provided by Dr. Scott Weaver. All virus strains were produced in Vero-E6 cells and then titrated, aliquoted and stored as previously reported [21].…”

“…HMC3 and HBEC-5i cells were grown in 12-well or 96-well plates and then infected with MAYV or UNAV at a multiplicity of infection (MOI) of 1 for a period of 24 to 72 h. After that, images were captured with an inverted microscope to analyze cell morphology. For the cell viability experiments, we used the MTT method as previously described [21]. After 24, 48 or 72 h of infection, 5 mg/ml of 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT, Sigma-Aldrich, St. Louis, MI, USA) solution was added to the cells and incubated for an additional 4 h. Formazan crystals were dissolved in a solution of 4 mM HCl and 10% Triton X-100 in isopropanol, and absorbance was determined at 570 nm using a microplate reader spectrophotometer (BioTeK, Winooski, VT, USA).…”

“…Virus progeny production in cell supernatants was determined using plaque-forming assays as previously performed [21]. For this end, 10-fold serial dilutions of MAYV- or UNAV-infected samples were used to infect confluent Vero-E6 cells grown in 6-well plates.…”

“…HCM3 or HBEC-5i cells grown on glass coverslips in 24-well plates were infected with MAYV or UNAV at an MOI of 1 for 48 h. Next, the cells were fixed, permeabilized, and blocked as previously reported [21]. After that, the cells were stained overnight at 4 °C with E1 or nsP1 primary rabbit antibodies (both antibodies previously validated in our laboratory) [22], followed by Alexa Flour 568 goat anti-rabbit secondary antibody (Invitrogen, Carlsbad, CA, USA).…”

An update on the development of antiviral against Mayaro virus: from molecules to potential viral targets

“…Finally, Valdés-Torres and collaborators investigated the potential of Honokiol and α-Mangostin against MAYV (Valdés-Torres et al 2022 ). Primary human dermal fibroblasts (HDFs), Vero-E6, and HeLa cells were infected with MAYV at an MOI of 1 for 1 h, and then treated with Honokiol (5 to 10 μM) or α-Mangostin (1 to 5 μM) for 24 h. Honokiol and α-Mangostin reduced MAYV-induced cytopathic effects in a dose-dependent manner and inhibited the production of viral progeny in pretreatment assay.…”