Expression of genes involved in immune response and in vitro immunosuppressive effect of equine MSCs.
Abstract: The immunomodulatory capacities of mesenchymal stem cells (MSCs) have made them the subject of increased clinical interest for tissue regeneration and repair. We have studied the immunomodulatory capacity of equine MSCs derived from bone marrow (BM-MSCs) and adipose tissue (AT-MSCs) in cocultures with allogeneic peripheral blood mononuclear cells (PBMCs). Different isoforms and concentrations of phytohaemaglutinin (PHA) were tested to determine the best stimulation conditions for PBMC proliferation and a proliferation assay was performed for 7 days to determine the optimal day of stimulation of PBMCs. The effect of the dose and source of MSCs was evaluated in cocultures of 10(5) PBMCs with different ratios of AT- and BM-MSCs (1:1, 1:10, 1:20 and 1:50). Proliferation rates of the PBMCs were evaluated using BrdU ELISA colorimetric assay. PHA stimulated equine PBMCs reached their peak of growth after 3 days of culture. The immunoassay showed a decrease of the PBMCs growth at high ratio cocultures (1:1 and 1:10). Equine BM-MSCs and AT-MSCs demonstrated an ability to suppress the proliferation of stimulated PBMCs. Although MSCs derived from both sources displayed immunosuppressive effects, AT-MSCs were slightly more potent than BM-MSCs. In addition, the expression of 26 genes coding for different molecules implicated in the immune response was analyzed in cocultures of BM-MSCs and PHA stimulated PBMSCs by reverse transcriptase real time quantitative PCR (RT-qPCR). An upregulation in genes associated with the production of interleukins and cytokines such as TNF-α and TGF-β1 was observed except for IFN-γ whose expression significantly decreased. The variations of interleukins and cytokine receptors showed no clear patterns. COX-1 and COX-2 showed similar expression patterns while INOs expression significantly decreased in the two cell types present in the coculture. Cyclin D2 and IDO-1 showed an increased expression and CD90, ITG-β1 and CD44 expression decreased significantly in BM-MSCs cocultured with PHA stimulated PBMCs. On the contrary, CD6 and VCAM1 expression increased in these cells. With regard to the expression of the five genes involved in antigen presentation, an upregulation was observed in both cocultured MSCs and stimulated PBMCs. This study contributes to the knowledge of the immunoregulatory properties of equine MSCs, which are notably important for the treatment of inflammation processes, such as tendinitis and osteoarthritis.
Copyright © 2015 Elsevier B.V. All rights reserved.
Publication Date: 2015-04-21 PubMed ID: 25977164DOI: 10.1016/j.vetimm.2015.04.004Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research paper explores the immunomodulatory capacities of equine mesenchymal stem cells (MSCs) derived from bone marrow and adipose tissue, by studying their interaction with allogeneic peripheral blood mononuclear cells (PBMCs). The study shows that both types of MSCs can suppress the proliferation of PBMCs, but adipose tissue MSCs were slightly more effective. The study also analyzed the expression of genes related to the immune response.
Understanding the Research
- This study focuses on the immunomodulatory capacities of equine MSCs collected from bone marrow (BM-MSCs) and adipose tissue (AT-MSCs). This stems from the growing interest in MSCs in the field of tissue regeneration and repair due to their ability to modulate the immune response.
- Preliminary testing was conducted in order to determine the best stimulation conditions for PBMC proliferation using different isoforms and concentrations of phytohaemaglutinin (PHA). Proliferation rates of the PBMCs were then tracked using a BrdU ELISA colorimetric assay.
- Once the optimal day of stimulation was determined, the effect of the dose and source of MSCs on PBMC proliferation was evaluated. PBMCs were cocultured with AT- and BM-MSCs at different ratios, with results indicating that PBMC growth decreases at high ratio cocultures.
- Equine BM-MSCs and AT-MSCs demonstrated their ability to suppress the proliferation of stimulated PBMCs though AT-MSCs were slightly more efficient.
Gene Expression Findings
- The study also investigated the expression of 26 genes associated with molecules involved in the immune response using reverse transcriptase real-time quantitative PCR (RT-qPCR).
- The genes evaluated included those associated with the production of interleukins, cytokines such as TNF-α and TGF-β1, COX-1, COX-2, INOs, Cyclin D2 and IDO-1, as well as genes involved in antigen presentation. Most showed an increase in expression except for the gene coding for IFN-γ, and INOs whose expression significantly decreased.
- Genes associated with the cell surface markers CD90, ITG-β1, and CD44 showed decreased expression while those for CD6 and VCAM1 indicated an increase when BM-MSCs were cocultured with PHA stimulated PBMCs.
Significance of the Findings
- The results from this study contribute greatly to the understanding of the immunoregulatory properties of equine MSCs. These findings could prove to be crucial in the treatment of inflammatory conditions such as tendinitis and osteoarthritis.
Cite This Article
APA
Remacha AR, Barrachina L, Álvarez-Arguedas S, Ranera B, Romero A, Vázquez FJ, Zaragoza P, Yañez R, Martín-Burriel I, Rodellar C.
(2015).
Expression of genes involved in immune response and in vitro immunosuppressive effect of equine MSCs.
Vet Immunol Immunopathol, 165(3-4), 107-118.
https://doi.org/10.1016/j.vetimm.2015.04.004 Publication
Researcher Affiliations
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain; Hospital Veterinario, Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain; Hospital Veterinario, Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
- Division of Hematopoietic Innovative Therapies, CIEMAT/CIBERER/FJD, Madrid, Spain.
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
- Laboratorio de Genética Bioquímica (LAGENBIO), Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain. Electronic address: rodellar@unizar.es.
MeSH Terms
- Animals
- Chemokine CXCL1 / physiology
- Genes, MHC Class II / genetics
- Horses / genetics
- Horses / immunology
- Immunosuppression Therapy / methods
- Immunosuppression Therapy / veterinary
- In Vitro Techniques
- Interferon-gamma / physiology
- Interleukin-10 / physiology
- Interleukin-6 / physiology
- Leukocytes, Mononuclear / immunology
- Leukocytes, Mononuclear / physiology
- Mesenchymal Stem Cells / immunology
- Mesenchymal Stem Cells / physiology
- Receptors, CXCR3 / physiology
- Receptors, CXCR4 / physiology
- Transcriptome / immunology
- Transforming Growth Factor beta1 / physiology
- Tumor Necrosis Factor-alpha / physiology
Citations
This article has been cited 14 times.- Phyo H, Aburza A, Mellanby K, Esteves CL. Characterization of canine adipose- and endometrium-derived Mesenchymal Stem/Stromal Cells and response to lipopolysaccharide. Front Vet Sci 2023;10:1180760.
- Cequier A, Vázquez FJ, Romero A, Vitoria A, Bernad E, García-Martínez M, Gascón I, Barrachina L, Rodellar C. The immunomodulation-immunogenicity balance of equine Mesenchymal Stem Cells (MSCs) is differentially affected by the immune cell response depending on inflammatory licensing and major histocompatibility complex (MHC) compatibility. Front Vet Sci 2022;9:957153.
- Cequier A, Romero A, Vázquez FJ, Vitoria A, Bernad E, Fuente S, Zaragoza P, Rodellar C, Barrachina L. Equine Mesenchymal Stem Cells Influence the Proliferative Response of Lymphocytes: Effect of Inflammation, Differentiation and MHC-Compatibility. Animals (Basel) 2022 Apr 11;12(8).
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- Shahsavari A, Weeratunga P, Ovchinnikov DA, Whitworth DJ. Pluripotency and immunomodulatory signatures of canine induced pluripotent stem cell-derived mesenchymal stromal cells are similar to harvested mesenchymal stromal cells. Sci Rep 2021 Feb 10;11(1):3486.
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- Yaneselli K, Ávila G, Rossi A, Rial A, Castro S, Estradé MJ, Suárez G, Algorta A. Impact of different formulations of platelet lysate on proliferative and immune profile of equine mesenchymal stromal cells. Front Vet Sci 2024;11:1410855.
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