Interferon-γ-Treated Mesenchymal Stem Cells Modulate the T Cell-Related Chemokines and Chemokine Receptors in an Animal Model of Experimental Autoimmune Encephalomyelitis

Abstract

Background Mesenchymal stem cells (MSCs) modulate immune responses, and their immunomodulatory potential can be enhanced using inflammatory cytokines. Here, the modulatory effects of IFN-γ-licensed MSCs on expression of T cell-related chemokines and chemokine receptors were evaluated using an experimental autoimmune encephalomyelitis (EAE) model.

Material and Methods EAE was induced in 3 groups of C57bl/6 mice and then treated with PBS, MSCs and IFN-γ-treated MSCs. The EAE manifestations were registered daily and finally, the brain and spinal cords were isolated for histopathological and gene expression studies.

Results The clinical scores were lowered in MSCs and IFN-γ-licensed MSCs groups, however, mice treated with IFN-γ-licensed MSCs exhibited lower clinical scores than MSCs-treated mice. Leukocyte infiltration into the brain was reduced after treatment with MSCs or IFN-γ-licensed MSCs compared to untreated group (P<0.05 and P<0.01, respectively). In comparison with untreated EAE mice, treatment with MSCs reduced CCL20 expression (P<0.001) and decreased CXCR3 and CCR6 expression (P<0.02 and P<0.04, respectively). In comparison with untreated EAE mice, treatment with IFN-γ-licensed MSCs reduced CXCL10, CCL17 and CCL20 expression (P<0.05, P<0.05, and P<0.001, respectively) as well as decreased CXCR3 and CCR6 expression (P<0.002 and P<0.02, respectively), whilst promoting expression of CCL22 and its receptor CCR4 (P<0.0001 and P<0.02, respectively). In comparison with MSC-treated group, mice treated with IFN-γ-licensed MSCs exhibited lower CXCL10 and CCR6 expression (P<0.002 and P<0.01, respectively), whereas greater expression of CCL22 and CCR4 (P<0.0001 and P<0.01, respectively).

Conclusion Priming the MSC with IFN-γ can be an efficient approach to enhance the immunomodulatory potential of MSCs.

Key words

Experimental autoimmune encephalomyelitis - IFN-γ-licensed MSCs - chemokines - chemokine receptors

Publication History

Received: 19 July 2022

Accepted: 28 November 2022

Article published online:
08 February 2023

• References

• 1 Kornek B, Lassmann H. Neuropathology of multiple sclerosis—new concepts. Brain Research Bulletin 2003; 61: 321-326 DOI: 10.1016/S0361-9230(03)00095-9.
• 2 Jafarzadeh A, Nemati M, Khorramdelazad H. et al. The Toll-like Receptor 2 (TLR2)-related Immunopathological Responses in the Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis. Iran J Allergy Asthma Immunol 2019; 18: 230-250 DOI: 10.18502/ijaai.v18i3.1117.
• 3 Ransohoff RM. Animal models of multiple sclerosis: the good, the bad and the bottom line. Nat Neurosci 2012; 15: 1074-1077 DOI: 10.1038/nn.3168.
• 4 Jafarzadeh A, Nemati M. Therapeutic potentials of ginger for treatment of Multiple sclerosis: A review with emphasis on its immunomodulatory, anti-inflammatory and anti-oxidative properties. J Neuroimmunol 2018; 324: 54-75 DOI: 10.1016/j.jneuroim.2018.09.003.
• 5 Etesam Z, Nemati M, Ebrahimizadeh MA. et al. Altered Expression of Specific Transcription Factors of Th17 (RORγt, RORα) and Treg Lymphocytes (FOXP3) by Peripheral Blood Mononuclear Cells from Patients with Multiple Sclerosis. J Mol Neurosci 2016; 60: 94-101 DOI: 10.1007/s12031-016-0789-5.
• 6 Greenwood J, Heasman SJ, Alvarez JI. et al. Review: Leucocyte–endothelial cell crosstalk at the blood–brain barrier: A prerequisite for successful immune cell entry to the brain. Neuropathology and Applied Neurobiology 2011; 37: 24-39 DOI: 10.1111/j.1365-2990.2010.01140.x.
• 7 Jafarzadeh A, Nemati M, Jafarzadeh S. The important role played by chemokines influence the clinical outcome of Helicobacter pylori infection. Life Sci 2019; 231: 116688 DOI: 10.1016/j.lfs.2019.116688.
• 8 Fallahi P, Ferrari SM, Ragusa F. et al. Th1 Chemokines in Autoimmune Endocrine Disorders. J Clin Endocrinol Metab 2020; 105 DOI: 10.1210/clinem/dgz289.
• 9 Jafarzadeh A, Azizi SV, Arabi Z. et al. Vitamin D down-regulates the expression of some Th17 cell-related cytokines, key inflammatory chemokines, and chemokine receptors in experimental autoimmune encephalomyelitis. Nutr Neurosci 2019; 22: 725-737 DOI: 10.1080/1028415x.2018.1436237.
• 10 Jafarzadeh A, Bagherzadeh S, Ebrahimi HA. et al. Higher circulating levels of chemokine CCL20 in patients with multiple sclerosis: evaluation of the influences of chemokine gene polymorphism, gender, treatment and disease pattern. J Mol Neurosci 2014; 53: 500-505 DOI: 10.1007/s12031-013-0214-2.
• 11 Scheu S, Ali S, Ruland C. et al. The C-C Chemokines CCL17 and CCL22 and Their Receptor CCR4 in CNS Autoimmunity. Int J Mol Sci 2017; 18: 2306 DOI: 10.3390/ijms18112306.
• 12 Jafarzadeh A, Ebrahimi HA, Bagherzadeh S. et al. Lower serum levels of Th2-related chemokine CCL22 in women patients with multiple sclerosis: a comparison between patients and healthy women. Inflammation 2014; 37: 604-610 DOI: 10.1007/s10753-013-9775-z.
• 13 Lee K, Majumdar MK, Buyaner D. et al. Human Mesenchymal Stem Cells Maintain Transgene Expression during Expansion and Differentiation. Molecular Therapy 2001; 3: 857-866 DOI: 10.1006/mthe.2001.0327.
• 14 Krampera M, Glennie S, Dyson J. et al. Bone marrow mesenchymal stem cells inhibit the response of naive and memory antigen-specific T cells to their cognate peptide. Blood 2003; 101: 3722-3729 DOI: 10.1182/blood-2002-07-2104.
• 15 Burt RK, Cohen BA, Russell E. et al. Hematopoietic stem cell transplantation for progressive multiple sclerosis: failure of a total body irradiation – based conditioning regimen to prevent disease progression in patients with high disability scores. Blood 2003; 102: 2373-2378 DOI: 10.1182/blood-2003-03-0877.
• 16 Draper JS, Smith K, Gokhale P. et al. Recurrent gain of chromosomes 17q and 12 in cultured human embryonic stem cells. Nature Biotechnology 2004; 22: 53-54 DOI: 10.1038/nbt922.
• 17 Augello A, Tasso R, Negrini SM. et al. Bone marrow mesenchymal progenitor cells inhibit lymphocyte proliferation by activation of the programmed death 1 pathway. European Journal of Immunology 2005; 35: 1482-1490 DOI: 10.1002/eji.200425405.
• 18 Aksu AE, Horibe E, Sacks J. et al. Co-infusion of donor bone marrow with host mesenchymal stem cells treats GVHD and promotes vascularized skin allograft survival in rats. Clin Immunol 2008; 127: 348-358 DOI: 10.1016/j.clim.2008.02.003.
• 19 Lim JH, Kim JS, Yoon IH. et al. Immunomodulation of delayed-type hypersensitivity responses by mesenchymal stem cells is associated with bystander T cell apoptosis in the draining lymph node. J Immunol 2010; 185: 4022-4029 DOI: 10.4049/jimmunol.0902723.
• 20 Ghannam S, Pène J, Moquet-Torcy G. et al. Mesenchymal stem cells inhibit human Th17 cell differentiation and function and induce a T regulatory cell phenotype. J Immunol 2010; 185: 302-312 DOI: 10.4049/jimmunol.0902007.
• 21 Matysiak M, Orlowski W, Fortak-Michalska M. et al. Immunoregulatory function of bone marrow mesenchymal stem cells in EAE depends on their differentiation state and secretion of PGE2. Journal of Neuroimmunology 2011; 233: 106-111 DOI: 10.1016/j.jneuroim.2010.12.004.
• 22 Coleman MA, Steptoe RJ. Induction of antigen-specific tolerance through hematopoietic stem cell-mediated gene therapy: the future for therapy of autoimmune disease. Autoimmun Rev 2012; 12: 195-203 DOI: 10.1016/j.autrev.2011.08.012.
• 23 Baker M. Stem cells and neurodegenerative disease: cool science and scepticism. Nature Reports Stem Cells 2009; DOI: 10.1038/stemcells.2009.54.
• 24 Lee BC, Kang KS. Functional enhancement strategies for immunomodulation of mesenchymal stem cells and their therapeutic application. Stem cell research & therapy 2020; 11: 397 DOI: 10.1186/s13287-020-01920-3.
• 25 English K, Barry FP, Field-Corbett CP. et al. IFN-γ and TNF-α differentially regulate immunomodulation by murine mesenchymal stem cells. Immunology Letters 2007; 110: 91-100 DOI: 10.1016/j.imlet.2007.04.001.
• 26 Chen L. Co-inhibitory molecules of the B7-CD28 family in the control of T-cell immunity. Nat Rev Immunol 2004; 4: 336-347 DOI: 10.1038/nri1349.
• 27 Polchert D, Sobinsky J, Douglas G. et al. IFN-γ activation of mesenchymal stem cells for treatment and prevention of graft versus host disease. European Journal of Immunology 2008; 38: 1745-1755 DOI: 10.1002/eji.200738129.
• 28 Ryan JM, Barry F, Murphy JM. et al. Interferon-gamma does not break, but promotes the immunosuppressive capacity of adult human mesenchymal stem cells. Clin Exp Immunol 2007; 149: 353-363 DOI: 10.1111/j.1365-2249.2007.03422.x.
• 29 Ren G, Zhang L, Zhao X. et al. Mesenchymal stem cell-mediated immunosuppression occurs via concerted action of chemokines and nitric oxide. Cell Stem Cell 2008; 2: 141-150 DOI: 10.1016/j.stem.2007.11.014.
• 30 Constantinescu CS, Farooqi N, O’Brien K. et al. Experimental autoimmune encephalomyelitis (EAE) as a model for multiple sclerosis (MS). British journal of pharmacology 2011; 164: 1079-1106 DOI: 10.1111/j.1476-5381.2011.01302.x.
• 31 Baker D, Amor S. Experimental autoimmune encephalomyelitis is a good model of multiple sclerosis if used wisely. Multiple sclerosis and related disorders 2014; 3: 555-564 DOI: 10.1016/j.msard.2014.05.002.
• 32 Bunnell BA, Flaat M, Gagliardi C. et al. Adipose-derived stem cells: isolation, expansion and differentiation. Methods 2008; 45: 115-120 DOI: 10.1016/j.ymeth.2008.03.006.
• 33 Rafei M, Birman E, Forner K. et al. Allogeneic mesenchymal stem cells for treatment of experimental autoimmune encephalomyelitis. Mol Ther 2009; 17: 1799-1803 DOI: 10.1038/mt.2009.157.
• 34 Ling X, Wang T, Han C. et al. IFN-γ-Primed hUCMSCs Significantly Reduced Inflammation via the Foxp3/ROR-γt/STAT3 Signaling Pathway in an Animal Model of Multiple Sclerosis. Frontiers in immunology 2022; 13: 835345-835345 DOI: 10.3389/fimmu.2022.835345.
• 35 Nozari P, Mokhtari P, Nemati M. et al. Investigation of the effect of IFN-γ/TNF-α-treated mesenchymal stem cells on Th9- and Treg cell-related parameters in a mouse model of ovalbumin-induced allergic asthma. Immunopharmacology and immunotoxicology 2022; 1-26 DOI: 10.1080/08923973.2022.2082977.
• 36 Takeshita K, Motoike S, Kajiya M. et al. Xenotransplantation of interferon-gamma-pretreated clumps of a human mesenchymal stem cell/extracellular matrix complex induces mouse calvarial bone regeneration. Stem cell research & therapy 2017; 8: 101-101 DOI: 10.1186/s13287-017-0550-1.
• 37 Rovira Gonzalez YI, Lynch PJ, Thompson EE. et al. In vitro cytokine licensing induces persistent permissive chromatin at the Indoleamine 2,3-dioxygenase promoter. Cytotherapy 2016; 18: 1114-1128 DOI: 10.1016/j.jcyt.2016.05.017.
• 38 Wang L, Zhao Y, Liu Y. et al. IFN-γ and TNF-α synergistically induce mesenchymal stem cell impairment and tumorigenesis via NFκB signaling. Stem cells 2013; 31: 1383-1395
• 39 Putra A, Ridwan FB, Putridewi AI. et al. The Role of TNF-α induced MSCs on Suppressive Inflammation by Increasing TGF-β and IL-10. Open Access Maced J Med Sci 2018; 6: 1779-1783 DOI: 10.3889/oamjms.2018.404.
• 40 Gimble JM, Bunnell BA, Chiu ES. et al. Concise review: Adipose-derived stromal vascular fraction cells and stem cells: let’s not get lost in translation. Stem Cells 2011; 29: 749-754 DOI: 10.1002/stem.629.
• 41 Zhang J, Li Y, Cui Y. et al. Erythropoietin treatment improves neurological functional recovery in EAE mice. Brain Research 2005; 1034: 34-39 DOI: 10.1016/j.brainres.2004.11.036.
• 42 Mix E, Meyer-Rienecker H, Hartung HP. et al. Animal models of multiple sclerosis--potentials and limitations. Prog Neurobiol 2010; 92: 386-404 DOI: 10.1016/j.pneurobio.2010.06.005.
• 43 Zhang J, Rong Y, Luo C. et al. Bone marrow mesenchymal stem cell-derived exosomes prevent osteoarthritis by regulating synovial macrophage polarization. Aging (Albany NY) 2020; 12: 25138-25152 DOI: 10.18632/aging.104110.
• 44 Kanai R, Nakashima A, Doi S. et al. Interferon-γ enhances the therapeutic effect of mesenchymal stem cells on experimental renal fibrosis. Sci Rep 2021; 11: 850 DOI: 10.1038/s41598-020-79664-6.
• 45 Uccelli A, Laroni A, Freedman MS. Mesenchymal stem cells for the treatment of multiple sclerosis and other neurological diseases. The Lancet Neurology 2011; 10: 649-656 DOI: 10.1016/S1474-4422(11)70121-1.
• 46 Saccardi R, Kozak T, Bocelli-Tyndall C. et al. Autologous stem cell transplantation for progressive multiple sclerosis: update of the European Group for Blood and Marrow Transplantation autoimmune diseases working party database. Mult Scler 2006; 12: 814-823 DOI: 10.1177/1352458506071301.
• 47 Musiał-Wysocka A, Kot M, Majka M. The Pros and Cons of Mesenchymal Stem Cell-Based Therapies. Cell Transplant 2019; 28: 801-812 DOI: 10.1177/0963689719837897.
• 48 Si Z, Wang X, Sun C. et al. Adipose-derived stem cells: Sources, potency, and implications for regenerative therapies. Biomedicine & Pharmacotherapy 2019; 114: 108765 DOI: 10.1016/j.biopha.2019.108765.
• 49 Tsuji W, Rubin JP, Marra KG. Adipose-derived stem cells: Implications in tissue regeneration. World J Stem Cells 2014; 6: 312-321 DOI: 10.4252/wjsc.v6.i3.312.
• 50 Li CY, Wu XY, Tong JB. et al. Comparative analysis of human mesenchymal stem cells from bone marrow and adipose tissue under xeno-free conditions for cell therapy. Stem cell research & therapy 2015; 6: 55 DOI: 10.1186/s13287-015-0066-5.
• 51 Torkaman M, Ghollasi M, Mohammadnia-Afrouzi M. et al. The effect of transplanted human Wharton’s jelly mesenchymal stem cells treated with IFN-γ on experimental autoimmune encephalomyelitis mice. Cellular immunology 2017; 311: 1-12 DOI: 10.1016/j.cellimm.2016.09.012.
• 52 Duijvestein M, Wildenberg ME, Welling MM. et al. Pretreatment with Interferon-γ Enhances the Therapeutic Activity of Mesenchymal Stromal Cells in Animal Models of Colitis. Stem Cells 2011; 29: 1549-1558 DOI: 10.1002/stem.698.
• 53 Kim DS, Jang IK, Lee MW. et al. Enhanced Immunosuppressive Properties of Human Mesenchymal Stem Cells Primed by Interferon-γ. EBioMedicine 2018; 28: 261-273 DOI: 10.1016/j.ebiom.2018.01.002.
• 54 Oh J-W, Schwiebert LM, Benveniste EN. Cytokine regulation of CC and CXC chemokine expression by human astrocytes. Journal of Neurovirology 1999; 5: 82-94 DOI: 10.3109/13550289909029749.
• 55 Li H, Wang W, Wang G. et al. Interferon-γ and tumor necrosis factor-α promote the ability of human placenta-derived mesenchymal stromal cells to express programmed death ligand-2 and induce the differentiation of CD4(+)interleukin-10(+) and CD8(+)interleukin-10(+)Treg subsets. Cytotherapy 2015; 17: 1560-1571 DOI: 10.1016/j.jcyt.2015.07.018.
• 56 Zappia E, Casazza S, Pedemonte E. et al. Mesenchymal stem cells ameliorate experimental autoimmune encephalomyelitis inducing T-cell anergy. Blood 2005; 106: 1755-1761 DOI: 10.1182/blood-2005-04-1496.
• 57 Rostami M, Haidari K, Amini H. et al. Genetically Engineered Mesenchymal Stem Cell Therapy Against Murine Experimental Autoimmune Encephalomyelitis. Molecular neurobiology 2022; 59: 3449-3457 DOI: 10.1007/s12035-022-02774-x.
• 58 Dang S, Xu H, Xu C. et al. Autophagy regulates the therapeutic potential of mesenchymal stem cells in experimental autoimmune encephalomyelitis. Autophagy 2014; 10: 1301-1315 DOI: 10.4161/auto.28771.
• 59 Chinnadurai R, Copland IB, Patel SR. et al. IDO-independent suppression of T cell effector function by IFN-γ-licensed human mesenchymal stromal cells. Journal of immunology (Baltimore, Md : 1950) 2014; 192: 1491-1501 DOI: 10.4049/jimmunol.1301828.
• 60 English K, Barry FP, Field-Corbett CP. et al. IFN-gamma and TNF-alpha differentially regulate immunomodulation by murine mesenchymal stem cells. Immunology letters 2007; 110: 91-100 DOI: 10.1016/j.imlet.2007.04.001.
• 61 Wright A, Arthaud-Day ML, Weiss ML. Therapeutic Use of Mesenchymal Stromal Cells: The Need for Inclusive Characterization Guidelines to Accommodate All Tissue Sources and Species. Frontiers in cell and developmental biology 2021; 9: 632717 DOI: 10.3389/fcell.2021.632717.