Cytokines Differently Define the Immunomodulation of Mesenchymal Stem Cells from the Periodontal Ligament
Abstract
1. Introduction
2. Materials and Methods
2.1. Ethics
2.2. hPDLSCs Culture and Verification of MSC Surface Marker Expression
2.3. CD4+ T Lymphocyte Isolation
2.4. Experimental Protocols
2.4.1. hPDLSCs Treatment
2.4.2. hPDLSCs/CD4+ T Cell Indirect Co-Culture
2.4.3. Immunomediators Expression
2.5. Analysis of Co-Culture Experiments
2.5.1. CD4+ T Cell Proliferation and Apoptosis
2.5.2. Immunomediators Expression Analysis in hPDLSCs
Quantitative Polymerase Chain Reaction (qPCR)
IDO-1 Immunostaining
Measuring L-Kynurenine Levels
PD-L1 and PD-L2 Immunostaining
Prostaglandin E2 ELISA
2.6. Statistical Analysis
3. Results
3.1. Mesenchymal and Hematopoietic Surface Marker Expression in hPDLSCs
3.2. hPDLSC Mediated Effect of Different Inflammatory Stimuli on CD4+ T Lymphocytes
3.3. Effect of Different Inflammatory Cytokines on the Expression of Immunomodulatory Proteins in hPDLSCs
3.3.1. IDO-1 Expression and Activity
3.3.2. PD-L1/2 Expression
3.3.3. PTGS-2/PGE2 Expression
3.4. Effect of IDO-1, PD-L1, and PTGS-2 Inhibition in hPDLSCs on CD4+ T Lymphocyte Proliferation in the Presence of Different Inflammatory Stimuli
3.5. Effect of IDO-1, PD-L1, and PTGS-2 Inhibition in hPDLSCs on CD4+ T Lymphocyte Apoptosis in the Presence of Different Inflammatory Stimuli
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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MSC Marker | Hematopoietic Marker | |
---|---|---|
CD29 | 97.7 ± 0.2 | - |
CD73 | 96.1 ± 0.2 | - |
CD90 | 97.9 ± 0.2 | - |
CD105 | 97.1 ± 0.6 | - |
CD146 | 61.3 ± 5.7 | - |
CD31 | - | 0.5 ± 0.1 |
CD34 | - | 0.6 ± 0.2 |
CD45 | - | 2.7 ± 0.2 |
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Behm, C.; Blufstein, A.; Gahn, J.; Nemec, M.; Moritz, A.; Rausch-Fan, X.; Andrukhov, O. Cytokines Differently Define the Immunomodulation of Mesenchymal Stem Cells from the Periodontal Ligament. Cells 2020, 9, 1222. https://doi.org/10.3390/cells9051222
Behm C, Blufstein A, Gahn J, Nemec M, Moritz A, Rausch-Fan X, Andrukhov O. Cytokines Differently Define the Immunomodulation of Mesenchymal Stem Cells from the Periodontal Ligament. Cells. 2020; 9(5):1222. https://doi.org/10.3390/cells9051222
Chicago/Turabian StyleBehm, Christian, Alice Blufstein, Johannes Gahn, Michael Nemec, Andreas Moritz, Xiaohui Rausch-Fan, and Oleh Andrukhov. 2020. "Cytokines Differently Define the Immunomodulation of Mesenchymal Stem Cells from the Periodontal Ligament" Cells 9, no. 5: 1222. https://doi.org/10.3390/cells9051222
APA StyleBehm, C., Blufstein, A., Gahn, J., Nemec, M., Moritz, A., Rausch-Fan, X., & Andrukhov, O. (2020). Cytokines Differently Define the Immunomodulation of Mesenchymal Stem Cells from the Periodontal Ligament. Cells, 9(5), 1222. https://doi.org/10.3390/cells9051222