Inhibition of p38 Mitogen-Activated Protein Kinase Impairs Mayaro Virus Replication in Human Dermal Fibroblasts and HeLa Cells.

Madelaine Sugasti-Salazar,Yessica Y Llamas-González,Dalkiria Campos,José González-Santamaría

doi:10.3390/v13061156

Madelaine Sugasti-Salazar, Yessica Y Llamas-González + Show 2 more

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https://doi.org/10.3390/v13061156

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Abstract

Mayaro virus (MAYV) hijacks the host’s cell machinery to effectively replicate. The mitogen-activated protein kinases (MAPKs) p38, JNK, and ERK1/2 have emerged as crucial cellular factors implicated in different stages of the viral cycle. However, whether MAYV uses these MAPKs to competently replicate has not yet been determined. The aim of this study was to evaluate the impact of MAPK inhibition on MAYV replication using primary human dermal fibroblasts (HDFs) and HeLa cells. Viral yields in supernatants from MAYV-infected cells treated or untreated with inhibitors SB203580, SP600125, U0126, or Losmapimod were quantified using plaque assay. Additionally, viral protein expression was analyzed using immunoblot and immunofluorescence. Knockdown of p38⍺/p38β isoforms was performed in HDFs using the PROTACs molecule NR-7h. Our data demonstrated that HDFs are highly susceptible to MAYV infection. SB203580, a p38 inhibitor, reduced MAYV replication in a dose-dependent manner in both HDFs and HeLa cells. Additionally, SB203580 significantly decreased viral E1 protein expression. Similarly, knockdown or inhibition of p38⍺/p38β isoforms with NR-7h or Losmapimod, respectively, affected MAYV replication in a dose-dependent manner. Collectively, these findings suggest that p38 could play an important role in MAYV replication and could serve as a therapeutic target to control MAYV infection.

Highlights

Mayaro virus (MAYV) is an emerging, neglected arbovirus belonging to the Togaviridae family within the Alphavirus genus [1,2]
In order to assess whether primary human dermal fibroblasts (HDFs) are susceptible to MAYV infection, we performed an infection kinetics experiment in HDFs with MAYV at an multiplicity of infection (MOI) of 1 and evaluated the cytopathic effect after viral infection
To determine whether the cytopathic effect observed was associated with an increase in the number of MAYV-infected cells, we decided to study the presence of MAYV antigens in infected HDFs at different times using an immunofluorescence confocal microscope

Summary

Introduction

Mayaro virus (MAYV) is an emerging, neglected arbovirus belonging to the Togaviridae family within the Alphavirus genus [1,2]. MAYV was isolated for the first time from the sera of forest workers in Trinidad and Tobago in 1954 [3]. Peru, French Guiana, Ecuador, Bolivia, Colombia, Venezuela, Haiti, Panama, and Trinidad and Tobago [4,5,6,7,8,9,10,11,12,13,14]. MAYV causes the disease known as Mayaro fever, which is characterized by nonspecific symptoms, including fever, myalgia, rash, headache, retro-orbital pain, diarrhea, vomiting, lymphadenopathy, leukopenia, and arthralgia [15]. MAYV is transmitted through the bites of sylvatic mosquitos, mainly members of the Haemagogus genus [15]. Several laboratory experiments suggest that urban mosquitos, such as Aedes aegypti or

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