Quercetin can reduce viral RNA level of O\u2019nyong-nyong virus and resulting innate immune cytokine responses in cultured human synovial fibroblasts

Axelle Septembre-Malaterre,Jimmy Sélambarom,Philippe Gasque,Yosra Bedoui,Pascale Guiraud,Claude Giry

doi:10.1038/s41598-021-85840-z

Axelle Septembre-Malaterre, Jimmy Sélambarom + Show 4 more

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https://doi.org/10.1038/s41598-021-85840-z

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Abstract

O’nyong-nyong virus is an alphavirus closely related to chikungunya virus, causing arthralgia, rash and fever. Alphaviruses mainly target synovial fibroblasts and persists in the joints of patients, possibly leading to chronic arthritis. To date, no specific antiviral treatment is available for ONNV infection and induced-inflammation. Primary human synovial fibroblasts cells were used to assess infection by ONNV and the resulting cytokine responses. Phenolics (gallic acid, caffeic acid and chlorogenic acid, curcumin and quercetin) and a curcuminoids-rich extract from turmeric were tested for their antiviral and anti-inflammatory capacities. We showed that infection occurred in HSF cells and increased gene expression and protein secretion of two major proinflammatory CCL-2 and IL-1β markers. In ONNV-infected HSF cells (MOI 1), we found that non-cytotoxic concentrations of phenolics (10 µM) reduced the level of viral RNA (E1, E2, nsP1, nsP2) and downregulated CCL-2 and IL-1β expression and secretion. These results highlighted the high value of the flavonol quercetin to reduce viral RNA levels and inflammatory status induced by ONNV in HSF cells.

Highlights

Abbreviations CaA Caffeic acid ChA Chlorogenic acid curcuminoid-rich extract (CRE) Curcuminoids-rich extract CU Curcumin chikungunya virus (CHIKV) Chinkungunya virus GaA Gallic acid DAPI 4′,6-Diamidino-2-phenylindole human synovial fibroblasts cells (HSF) Human synovial fibroblasts lactate dehydrogenase (LDH) Lactate dehydrogenase MOI Multiplicity of infection ONNV O’nyong-nyong virus PFU Plaque Forming Unit QU Quercetin RT-PCR Real-time polymerase chain reaction qRT-PCR Quantitative RT-PCR ELISA Enzyme-linked immunosorbent assay
Escalating MOI ( 10–3 to 1) of ONNV did not significantly affect LDH release from HSF cells when compared to the basal release (CTRL)
Significant LDH release compared to untreated cells (CTRL) was observed for the highest doses of gallic acid (100 μM; p < 0.01), caffeic acid (> 25 μM; p < 0.001), chlorogenic acid (> 50 μM; p < 0.01 and 0.001), quercetin (> 50 μM; p < 0.01 and 0.001) and curcumin (100 μM; p < 0.01), while CRE led to an insignificant effect on LDH release for all the tested concentrations

Summary

Introduction

Abbreviations CaA Caffeic acid ChA Chlorogenic acid CRE Curcuminoids-rich extract CU Curcumin CHIKV Chinkungunya virus GaA Gallic acid DAPI 4′,6-Diamidino-2-phenylindole HSF Human synovial fibroblasts LDH Lactate dehydrogenase MOI Multiplicity of infection ONNV O’nyong-nyong virus PFU Plaque Forming Unit QU Quercetin RT-PCR Real-time polymerase chain reaction qRT-PCR Quantitative RT-PCR ELISA Enzyme-linked immunosorbent assay. O’nyong-nyong virus (ONNV) is a mosquito-borne virus belonging to the genus Alphavirus in the Togaviridae family This RNA virus, closely related antigenically, genetically and clinically to chikungunya virus (CHIKV), Scientific Reports | (2021) 11:6369. ONNV causes symptoms similar to those caused by CHIKV including fever, rash and a rthralgia[6,7,8] Due to their antigenic and clinical similarities, the development of vaccines or drugs may serve in both cases[9], as recently reported with a serogrouping-protection vaccine using mouse models[10]. Synovial fibroblasts are enrolled in cell signaling via interactions with other major immune cells including monocytes and macrophages[17] These synovial-derived cells have gained growing interest during the past decade to study in vitro alphavirus infection and management, with a recent attempt to clarify their r ole[18]

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