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Stem cell research & therapy2014; 5(1); 13; doi: 10.1186/scrt402

Equine bone marrow-derived mesenchymal stromal cells are heterogeneous in MHC class II expression and capable of inciting an immune response in vitro.

Abstract: The horse is a valuable species to assess the effect of allogeneic mesenchymal stromal cells (MSCs) in regenerative treatments. No studies to date have examined recipient response to major histocompatibility complex (MHC)-mismatched equine MSCs. The purposes of this study were to immunophenotype MSCs from horses of known MHC haplotype and to compare the immunogenicity of MSCs with differing MHC class II expression. Methods: MSCs and peripheral blood leukocytes (PBLs) were obtained from Thoroughbred horses (n=10) of known MHC haplotype (ELA-A2, -A3, and -A9 homozygotes). MSCs were cultured through P8; cells from each passage (P2 to P8) were cryopreserved until used. Immunophenotyping of MHC class I and II, CD44, CD29, CD90, LFA-1, and CD45RB was performed by using flow cytometry. Tri-lineage differentiation assays were performed to confirm MSC multipotency. Recombinant equine IFN-γ was used to stimulate MHC class II negative MSCs in culture, after which expression of MHC class II was re-examined. To assess the ability of MHC class II negative or positive MSCs to stimulate an immune response, modified one-way mixed leukocyte reactions (MLRs) were performed by using MHC-matched and mismatched responder PBLs and stimulator PBLs or MSCs. Proliferation of gated CFSE-labeled CD3+ responder T cells was evaluated via CFSE attenuation by using flow cytometry and reported as the number of cells in the proliferating T-cell gate. Results: MSCs varied widely in MHC class II expression despite being homogenous in terms of "stemness" marker expression and ability to undergo trilineage differentiation. Stimulation of MHC class II negative MSCs with IFN-γ resulted in markedly increased expression of MHC class II. MLR results revealed that MHC-mismatched MHC class II-positive MSCs caused significantly increased responder T-cell proliferation compared with MHC-mismatched MHC class II-negative and MHC-matched MSCs, and equivalent to that of the positive control of MHC-mismatched leukocytes. Conclusions: The results of this study suggest that MSCs should be confirmed as MHC class II negative before allogeneic application. Additionally, it must be considered that even MHC class II-negative MSCs could upregulate MHC class II expression if implanted into an area of active inflammation, as demonstrated with in vitro stimulation with IFN-γ.
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Publication Date: 2014-01-24 PubMed ID: 24461709PubMed Central: PMC4055004DOI: 10.1186/scrt402Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research explores how horse mesenchymal stromal cells (MSCs), used in regenerative treatments, can incite an immune response. The team examined if variations in major histocompatibility complex (MHC) class II expression might be the cause and concluded that only MSCs confirmed as MHC class II negative should be used in treatment.

Research Objectives and Methodology

  • The aim of this study was two-fold: Firstly, to immunophenotype MSCs from horses whose MHC haplotype was known and, secondly, to compare the immunogenicity of MSCs with varying MHC class II expression.
  • MSCs and peripheral blood leukocytes (PBLs) were collected and cultured from ten thoroughbred horses, and cells from each passage stage were preserved for analysis.
  • The team used flow cytometry to immunophenotype crucial markers: MHC class I and II, CD44, CD29, CD90, LFA-1, and CD45RB.
  • To confirm the multipotency of the MSCs, tri-lineage differentiation assays were performed.
  • The researchers used recombinant equine IFN-γ to stimulate MHC class II negative MSCs in culture. Once stimulated, the MHC class II expression was evaluated again.
  • Lastly, to assess the ability of MHC class II negative or positive MSCs to stimulate an immune response, modified one-way mixed leukocyte reactions (MLRs) were performed.

Results of the Research

  • Despite the MSCs being homogeneous in terms of “stemness” marker expression, the study found significant variations in MHC class II expression.
  • Stimulation of MHC class II negative MSCs with IFN-γ resulted in a significant increase in MHC class II expression.
  • MLR results highlighted that MHC class II-positive MSCs, when MHC-mismatched, caused an increased T-cell response compared to MHC class II-negative and MHC-matched MSCs. The response was similar to the positive control of MHC-mismatched leukocytes.

Conclusions of the Research

  • The research concludes that for allogeneic applications, only MSCs confirmed as MHC class II negative should be used.
  • The researchers also warn that even MHC class II-negative MSCs could potentially upregulate MHC class II expression if implanted into a region with active inflammation. This conclusion was based on the in-vitro stimulation with IFN-γ.

Cite This Article

APA
Schnabel LV, Pezzanite LM, Antczak DF, Felippe MJ, Fortier LA. (2014). Equine bone marrow-derived mesenchymal stromal cells are heterogeneous in MHC class II expression and capable of inciting an immune response in vitro. Stem Cell Res Ther, 5(1), 13. https://doi.org/10.1186/scrt402

Publication

Stem cell research & therapy
ISSN: 1757-6512
NlmUniqueID: 101527581
Country: England
Language: English
Volume: 5
Issue: 1
Pages: 13

Researcher Affiliations

Schnabel, Lauren V
    Pezzanite, Lynn M
      Antczak, Douglas F
        Felippe, M Julia Bevilaqua
          Fortier, Lisa A

            MeSH Terms

            • Animals
            • Cells, Cultured
            • Genes, MHC Class II / genetics
            • Genes, MHC Class II / immunology
            • Horses
            • Immunophenotyping
            • Interferon-gamma / pharmacology
            • Leukocytes / immunology
            • Mesenchymal Stem Cells / cytology
            • Mesenchymal Stem Cells / drug effects
            • Mesenchymal Stem Cells / immunology

            Grant Funding

            • K08 AR060875 / NIAMS NIH HHS
            • 1K08AR060875-01A1 / NIAMS NIH HHS
            • S10RR023781 / NCRR NIH HHS

            References

            This article includes 41 references
            1. Griffin MD, Ryan AE, Alagesan S, Lohan P, Treacy O, Ritter T. Anti-donor immune responses elicited by allogeneic mesenchymal stem cells: what have we learned so far?. Immunol Cell Biol 2013 Jan;91(1):40-51.
              doi: 10.1038/icb.2012.67pubmed: 23207278google scholar: lookup
            2. Stagg J, Galipeau J. Immune plasticity of bone marrow-derived mesenchymal stromal cells.. Handb Exp Pharmacol 2007;(180):45-66.
              doi: 10.1007/978-3-540-68976-8_3pubmed: 17554504google scholar: lookup
            3. Stagg J. Immune regulation by mesenchymal stem cells: two sides to the coin.. Tissue Antigens 2007 Jan;69(1):1-9.
              pubmed: 17212702doi: 10.1111/j.1399-0039.2006.00739.xgoogle scholar: lookup
            4. Griffin MD, Ritter T, Mahon BP. Immunological aspects of allogeneic mesenchymal stem cell therapies.. Hum Gene Ther 2010 Dec;21(12):1641-55.
              doi: 10.1089/hum.2010.156pubmed: 20718666google scholar: lookup
            5. Eliopoulos N, Stagg J, Lejeune L, Pommey S, Galipeau J. Allogeneic marrow stromal cells are immune rejected by MHC class I- and class II-mismatched recipient mice.. Blood 2005 Dec 15;106(13):4057-65.
              doi: 10.1182/blood-2005-03-1004pubmed: 16118325google scholar: lookup
            6. Potian JA, Aviv H, Ponzio NM, Harrison JS, Rameshwar P. Veto-like activity of mesenchymal stem cells: functional discrimination between cellular responses to alloantigens and recall antigens.. J Immunol 2003 Oct 1;171(7):3426-34.
              pubmed: 14500637doi: 10.4049/jimmunol.171.7.3426google scholar: lookup
            7. Zangi L, Margalit R, Reich-Zeliger S, Bachar-Lustig E, Beilhack A, Negrin R, Reisner Y. Direct imaging of immune rejection and memory induction by allogeneic mesenchymal stromal cells.. Stem Cells 2009 Nov;27(11):2865-74.
              doi: 10.1002/stem.217pubmed: 19750539google scholar: lookup
            8. Badillo AT, Beggs KJ, Javazon EH, Tebbets JC, Flake AW. Murine bone marrow stromal progenitor cells elicit an in vivo cellular and humoral alloimmune response.. Biol Blood Marrow Transplant 2007 Apr;13(4):412-22.
              doi: 10.1016/j.bbmt.2006.12.447pmc: PMC1892590pubmed: 17382248google scholar: lookup
            9. Nauta AJ, Westerhuis G, Kruisselbrink AB, Lurvink EG, Willemze R, Fibbe WE. Donor-derived mesenchymal stem cells are immunogenic in an allogeneic host and stimulate donor graft rejection in a nonmyeloablative setting.. Blood 2006 Sep 15;108(6):2114-20.
              doi: 10.1182/blood-2005-11-011650pmc: PMC1895546pubmed: 16690970google scholar: lookup
            10. Jones EA, Kinsey SE, English A, Jones RA, Straszynski L, Meredith DM, Markham AF, Jack A, Emery P, McGonagle D. Isolation and characterization of bone marrow multipotential mesenchymal progenitor cells.. Arthritis Rheum 2002 Dec;46(12):3349-60.
              doi: 10.1002/art.10696pubmed: 12483742google scholar: lookup
            11. Fortier LA. Making progress in the what, when and where of regenerative medicine for our equine patients.. Equine Vet J 2012 Sep;44(5):511-2.
              doi: 10.1111/j.2042-3306.2012.00628.xpubmed: 22888773google scholar: lookup
            12. Fortier LA, Travis AJ. Stem cells in veterinary medicine.. Stem Cell Res Ther 2011 Feb 23;2(1):9.
              doi: 10.1186/scrt50pmc: PMC3092149pubmed: 21371354google scholar: lookup
            13. Frisbie DD, Smith RK. Clinical update on the use of mesenchymal stem cells in equine orthopaedics.. Equine Vet J 2010 Jan;42(1):86-9.
              pubmed: 20121921doi: 10.2746/042516409X477263google scholar: lookup
            14. Frisbie DD, Stewart MC. Cell-based therapies for equine joint disease.. Vet Clin North Am Equine Pract 2011 Aug;27(2):335-49.
              doi: 10.1016/j.cveq.2011.06.005pubmed: 21872762google scholar: lookup
            15. Schnabel LV, Fortier LA, McIlwraith CW, Nobert KM. Therapeutic use of stem cells in horses: which type, how, and when?. Vet J 2013 Sep;197(3):570-7.
              doi: 10.1016/j.tvjl.2013.04.018pubmed: 23778257google scholar: lookup
            16. De Schauwer C, Van de Walle GR, Van Soom A, Meyer E. Mesenchymal stem cell therapy in horses: useful beyond orthopedic injuries?. Vet Q 2013 Dec;33(4):234-41.
              pubmed: 23697553doi: 10.1080/01652176.2013.800250google scholar: lookup
            17. Guest DJ, Ousey JC, Smith MR. Defining the expression of marker genes in equine mesenchymal stromal cells.. Stem Cells Cloning 2008;1:1-9.
              pmc: PMC3781685pubmed: 24198500doi: 10.2147/sccaa.s3824google scholar: lookup
            18. De Schauwer C, Meyer E, Van de Walle GR, Van Soom A. Markers of stemness in equine mesenchymal stem cells: a plea for uniformity.. Theriogenology 2011 May;75(8):1431-43.
              doi: 10.1016/j.theriogenology.2010.11.008pubmed: 21196039google scholar: lookup
            19. Carrade DD, Lame MW, Kent MS, Clark KC, Walker NJ, Borjesson DL. Comparative Analysis of the Immunomodulatory Properties of Equine Adult-Derived Mesenchymal Stem Cells().. Cell Med 2012;4(1):1-11.
              doi: 10.3727/215517912X647217pmc: PMC3495591pubmed: 23152950google scholar: lookup
            20. Guest DJ, Smith MR, Allen WR. Monitoring the fate of autologous and allogeneic mesenchymal progenitor cells injected into the superficial digital flexor tendon of horses: preliminary study.. Equine Vet J 2008 Mar;40(2):178-81.
              doi: 10.2746/042516408X276942pubmed: 18267891google scholar: lookup
            21. Schnabel LV, Lynch ME, van der Meulen MC, Yeager AE, Kornatowski MA, Nixon AJ. Mesenchymal stem cells and insulin-like growth factor-I gene-enhanced mesenchymal stem cells improve structural aspects of healing in equine flexor digitorum superficialis tendons.. J Orthop Res 2009 Oct;27(10):1392-8.
              doi: 10.1002/jor.20887pubmed: 19350658google scholar: lookup
            22. Godwin EE, Young NJ, Dudhia J, Beamish IC, Smith RK. Implantation of bone marrow-derived mesenchymal stem cells demonstrates improved outcome in horses with overstrain injury of the superficial digital flexor tendon.. Equine Vet J 2012 Jan;44(1):25-32.
              doi: 10.1111/j.2042-3306.2011.00363.xpubmed: 21615465google scholar: lookup
            23. Wilke MM, Nydam DV, Nixon AJ. Enhanced early chondrogenesis in articular defects following arthroscopic mesenchymal stem cell implantation in an equine model.. J Orthop Res 2007 Jul;25(7):913-25.
              doi: 10.1002/jor.20382pubmed: 17405160google scholar: lookup
            24. Ferris DJ, Frisbie DD, Kisiday JD, McIlwraith CW, Hague BA, Major MD, Schneider RK, Zubrod CJ, Kawcak CE, Goodrich LR. Clinical outcome after intra-articular administration of bone marrow derived mesenchymal stem cells in 33 horses with stifle injury.. Vet Surg 2014 Mar;43(3):255-65.
              pubmed: 24433318doi: 10.1111/j.1532-950X.2014.12100.xgoogle scholar: lookup
            25. Frisbie DD, Kisiday JD, Kawcak CE, Werpy NM, McIlwraith CW. Evaluation of adipose-derived stromal vascular fraction or bone marrow-derived mesenchymal stem cells for treatment of osteoarthritis.. J Orthop Res 2009 Dec;27(12):1675-80.
              doi: 10.1002/jor.20933pubmed: 19544397google scholar: lookup
            26. McIlwraith CW, Frisbie DD, Rodkey WG, Kisiday JD, Werpy NM, Kawcak CE, Steadman JR. Evaluation of intra-articular mesenchymal stem cells to augment healing of microfractured chondral defects.. Arthroscopy 2011 Nov;27(11):1552-61.
              doi: 10.1016/j.arthro.2011.06.002pubmed: 21862278google scholar: lookup
            27. Lazary S, Antczak DF, Bailey E, Bell TK, Bernoco D, Byrns G, McClure JJ. Joint Report of the Fifth International Workshop on Lymphocyte Alloantigens of the Horse, Baton Rouge, Louisiana, 31 October-1 November 1987.. Anim Genet 1988;19(4):447-56.
              pubmed: 2466424doi: 10.1111/j.1365-2052.1988.tb00836.xgoogle scholar: lookup
            28. Tallmadge RL, Campbell JA, Miller DC, Antczak DF. Analysis of MHC class I genes across horse MHC haplotypes.. Immunogenetics 2010 Mar;62(3):159-72.
              doi: 10.1007/s00251-009-0420-9pmc: PMC2872545pubmed: 20099063google scholar: lookup
            29. Tallmadge RL, Lear TL, Antczak DF. Genomic characterization of MHC class I genes of the horse.. Immunogenetics 2005 Nov;57(10):763-74.
              doi: 10.1007/s00251-005-0034-9pubmed: 16220348google scholar: lookup
            30. Tseng CT, Miller D, Cassano J, Bailey E, Antczak DF. Identification of equine major histocompatibility complex haplotypes using polymorphic microsatellites.. Anim Genet 2010 Dec;41 Suppl 2(Suppl 2):150-3.
              pmc: PMC3289534pubmed: 21070289doi: 10.1111/j.1365-2052.2010.02125.xgoogle scholar: lookup
            31. Flaminio MJ, Antczak DF. Inhibition of lymphocyte proliferation and activation: a mechanism used by equine invasive trophoblast to escape the maternal immune response.. Placenta 2005 Feb-Mar;26(2-3):148-59.
              doi: 10.1016/j.placenta.2004.05.008pubmed: 15708116google scholar: lookup
            32. Radcliffe CH, Flaminio MJ, Fortier LA. Temporal analysis of equine bone marrow aspirate during establishment of putative mesenchymal progenitor cell populations.. Stem Cells Dev 2010 Feb;19(2):269-82.
              doi: 10.1089/scd.2009.0091pmc: PMC3138180pubmed: 19604071google scholar: lookup
            33. Watts AE, Ackerman-Yost JC, Nixon AJ. A comparison of three-dimensional culture systems to evaluate in vitro chondrogenesis of equine bone marrow-derived mesenchymal stem cells.. Tissue Eng Part A 2013 Oct;19(19-20):2275-83.
              doi: 10.1089/ten.tea.2012.0479pmc: PMC3761431pubmed: 23725547google scholar: lookup
            34. Chan WK, Lau AS, Li JC, Law HK, Lau YL, Chan GC. MHC expression kinetics and immunogenicity of mesenchymal stromal cells after short-term IFN-gamma challenge.. Exp Hematol 2008 Nov;36(11):1545-55.
              doi: 10.1016/j.exphem.2008.06.008pubmed: 18715686google scholar: lookup
            35. Romieu-Mourez R, François M, Boivin MN, Stagg J, Galipeau J. Regulation of MHC class II expression and antigen processing in murine and human mesenchymal stromal cells by IFN-gamma, TGF-beta, and cell density.. J Immunol 2007 Aug 1;179(3):1549-58.
              pubmed: 17641021doi: 10.4049/jimmunol.179.3.1549google scholar: lookup
            36. Crovace A, Lacitignola L, De Siena R, Rossi G, Francioso E. Cell therapy for tendon repair in horses: an experimental study.. Vet Res Commun 2007 Aug;31 Suppl 1:281-3.
              doi: 10.1007/s11259-007-0047-ypubmed: 17682895google scholar: lookup
            37. Crovace A, Lacitignola L, Rossi G, Francioso E. Histological and immunohistochemical evaluation of autologous cultured bone marrow mesenchymal stem cells and bone marrow mononucleated cells in collagenase-induced tendinitis of equine superficial digital flexor tendon.. Vet Med Int 2010;2010:250978.
              pmc: PMC2859019pubmed: 20445779doi: 10.4061/2010/250978google scholar: lookup
            38. Antczak DF, Bailey E, Barger B, Guerin G, Lazary S, McClure J, Mottironi VD, Symons R, Templeton J, Varewyck H. Joint report of the Third International Workshop on Lymphocyte Alloantigens of the Horse, Kennett Square, Pennsylvania, 25-27 April 1984.. Anim Genet 1986;17(4):363-73.
              pubmed: 3826760doi: 10.1111/j.1365-2052.1986.tb00730.xgoogle scholar: lookup
            39. Delling U, Lindner K, Ribitsch I, Jülke H, Brehm W. Comparison of bone marrow aspiration at the sternum and the tuber coxae in middle-aged horses.. Can J Vet Res 2012 Jan;76(1):52-6.
              pmc: PMC3244288pubmed: 22754095
            40. Baxter MA, Wynn RF, Jowitt SN, Wraith JE, Fairbairn LJ, Bellantuono I. Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion.. Stem Cells 2004;22(5):675-82.
              doi: 10.1634/stemcells.22-5-675pubmed: 15342932google scholar: lookup
            41. Majors AK, Boehm CA, Nitto H, Midura RJ, Muschler GF. Characterization of human bone marrow stromal cells with respect to osteoblastic differentiation.. J Orthop Res 1997 Jul;15(4):546-57.
              doi: 10.1002/jor.1100150410pubmed: 9379264google scholar: lookup

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            This article has been cited 77 times.
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