Human Synovial Mesenchymal Stem Cells Expressed Immunoregulatory Factors IDO and TSG6 in a Context of Arthritis Mediated by Alphaviruses
Abstract
:1. Introduction
2. Results
2.1. ONNV Can Infect Human Synovial Tissue-Derived MSCs
2.2. ONNV (MOI of 1) Alone or in Combination with Pro-Inflammatory Mediators Was Not Cytotoxic to Human Synovial MSCs and Did Not Affect Cell Proliferation
2.3. ONNV (MOI of 1) Alone or in Combination with Pro-Inflammatory Cytokines Promoted the Expression of the Antiviral ISG15 mRNA
2.4. ONNV (MOI of 1) Alone Failed to Upregulate the Expression of Pro-Inflammatory Factors (IL1β, IL6, CCL2)
2.5. ONNV Alone and/or in the Presence of Pro-Inflammatory Mediators Can Upregulate the Expression of Immunoregulatory Factors IDO and TSG6 by Human Synovial MSCs
2.6. ONNV Infection Increases GAS6 Expression but Has No Effect on PROS1 Expression in Human Synovial Tissue-Derived MSCs
2.7. There Was No Correlation between Pro- and Anti-Inflammatory Responses
2.8. ONNV Infection Downregulated the Expression of Immunoregulatory miR-146a-5p in Human Synovial MSCs Treated with TNF-α
3. Discussion
4. Materials and Methods
4.1. Cell Culture Reagents and Viruses
4.2. Cell Treatments
4.3. Cytotoxicity Assay
4.4. Mitochondrial Metabolic Activity Measurements
4.5. RNA Extraction and qPCR
4.6. mi-RNA Extraction and RT-PCR
4.7. Immunofluorescence
4.8. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Target Gene | Forward Sequence (5′–3′) | Reverse Sequence (3′–5′) |
---|---|---|
GAPDH | TGCGTCGCCAGCCGAG | AGTTAAAAGCAGCCCTGGTGA |
GAS6 | ACGACCCCGAGACGGATTAT | CTTCCTATCGCAGGGGTTGG |
IDO | AGCCCCTGACTTATGAGAACA | AGCTATTTCCAACAGCGCCT |
MXRA8 | TACTGTGGCCTGCACGAAC | CTCTCGGGGACGATGACATT |
ONNV Capsid | TAGAACACGCCCGTCGTATG | GCAACGCCTTCAGAAACGC |
ONNV E1 | CACCGTCCCCGTACGTAAAA | GGCTCTGTAGGCTGATGCAA |
ONNV E2 | CCCCTGACTACACGCTGATG | CCTTCATTGGAGCCGTCACA |
PROS1 | GTCTCAGAGGCAAACTTTTGTT | AGAATTTGCACGACGCTTCC |
TSG6 | TGGCTTTGTGGGAAGATACYGT | TGGAAACCTCCAGCTGTCAC |
Target mi-RNAs | Forward (5′–3′) |
---|---|
hsa-miR-146a-5p | GCAGTGAGAACTGAATTCCATG |
hsa-miR-221-3p | GCAGAGCTACATTGTCTGCT |
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Payet, M.; Septembre-Malaterre, A.; Gasque, P.; Guillot, X. Human Synovial Mesenchymal Stem Cells Expressed Immunoregulatory Factors IDO and TSG6 in a Context of Arthritis Mediated by Alphaviruses. Int. J. Mol. Sci. 2023, 24, 15932. https://doi.org/10.3390/ijms242115932
Payet M, Septembre-Malaterre A, Gasque P, Guillot X. Human Synovial Mesenchymal Stem Cells Expressed Immunoregulatory Factors IDO and TSG6 in a Context of Arthritis Mediated by Alphaviruses. International Journal of Molecular Sciences. 2023; 24(21):15932. https://doi.org/10.3390/ijms242115932
Chicago/Turabian StylePayet, Melissa, Axelle Septembre-Malaterre, Philippe Gasque, and Xavier Guillot. 2023. "Human Synovial Mesenchymal Stem Cells Expressed Immunoregulatory Factors IDO and TSG6 in a Context of Arthritis Mediated by Alphaviruses" International Journal of Molecular Sciences 24, no. 21: 15932. https://doi.org/10.3390/ijms242115932