Immunomodulatory drug methotrexate used to treat patients with chronic inflammatory rheumatisms post-chikungunya does not impair the synovial antiviral and bone repair responses.

Yosra Bedoui,Philippe Gasque,Marie-Christine Jaffar-Bandjee,Jimmy Selambarom,Pascale Guiraud,Claude Giry

doi:10.1371/journal.pntd.0006634

Abstract

Chikungunya virus (CHIKV) is a mosquito-transmitted RNA alphavirus causing major outbreaks of infectious chronic inflammatory rheumatisms (CIR). Recently, methotrexate (MTX), a disease modifying anti-rheumatic drug has been used successfully to treat patients suffering from rheumatoid-like arthritis post-CHIK but its immunomodulatory activity in the context of viral persistence has been a matter of concerns. We herein used a model of primary human synovial fibroblasts (HSF) and the synthetic molecule polyriboinosinic:polyribocytidylic acid (PIC) to mimic chronic infectious settings in the joints of CHIKV infected patients. The innate antiviral immune and inflammatory responses were investigated in response to MTX used at the therapeutic concentration of 1 μM. We found that MTX did not affect cellular viability as indicated by the LDH release assay. By quantitative RT-PCR, we observed that HSF responded robustly to PIC by increasing ISG15 and IFNβ mRNA levels. Furthermore, PIC upregulated the mRNA expression of two of the major pattern recognition receptors, RIG-I and MDA5 involved in the innate immune detection of viral RNA. MTX did not impact the antiviral response of PIC on ISG15, IFNβ, RIG-I and MDA5 mRNA expressions. MTX alone or combined with PIC did not affect the expression of proinflammatory CCL2 and CXCL8 chemokines. PIC strongly upregulated the mRNA and protein expression of osteoclastogenic factors (IL-6, GM-CSF but not RANKL). Critically, MTX treatment alone or combined with PIC did not affect the expression of all three tested osteoclastogenic cytokines. We found that MTX alone did not increase the capacity of CHIKV to infect and replicate in HSF. In conclusion, our study argues for a beneficial effect of MTX to treat CIR post-CHIKV given that it does not critically impact the antiviral, the proinflammatory and the bone tissue remodeling responses of synovial cells.

Highlights

Alphaviruses, transmitted by bites of infected mosquitoes, are globally distributed and capable to cause significant inflammatory diseases including arthritis and encephalitis [1]
Using a model of primary human joint fibroblasts (HSF), we investigated the capacity of the MTX immunosuppressive drug to affect the immune antiviral and inflammatory responses essential to clear the virus while allowing bone tissue repair
This study is important given that Chikungunya virus (CHIKV) and its RNA were shown to persist in the joint for months to years post infection and leading to injuries through ill-characterized mechanisms

Summary

Introduction

Alphaviruses, transmitted by bites of infected mosquitoes, are globally distributed and capable to cause significant inflammatory diseases including arthritis and encephalitis [1]. Old World alphaviruses, such as Chikungunya virus (CHIKV), Ross River virus (RRV) and O’NyongNyong virus (ONNV), are associated with rheumatic diseases in humans which can be chronic and severely debilitating [2]. Severe complications in adults such as persistent arthralgia and destructive arthritis have been reported, consistent with chronic inflammatory rheumatisms (CIR)[5,6,7,8]. The immunopathological mechanisms responsible for CHIK-CIR are poorly understood but may be due to viral persistence leading to chronic expression of viral RNA and with local inflammatory responses to drive osteoclastogenic activities [9,10,11,12]

Methods

Results

Discussion

Conclusion