Summary
Implantation of skeletal muscle precursor cells is a potential means of cell-mediated gene therapy. One unresolved question is the degree of immunogenicity of such myoblasts. We designed the extreme situation of implanting cells of a non-histocompatible myoblast cell line into cryodamaged, but regeneration-capable, muscles of adult mice. Without immuno-suppression donor cells are rejected within the first weeks. Immunosuppression with Cyclosporin A prevented invasion of T-lymphocytes and allowed differentiation of implanted myoblasts into myofibres as well as down-regulation of MHC expression. Still, withdrawal of Cyclosporin A after 4 weeks triggered lymphocyte invasion and cytotoxic cell reactions with rejection of donor tissue. Although the vast majority of muscle fibres was MHC-negative 1–4 days after Cyclosporin A withdrawal, single small desmin-positive profiles were weakly positive for donor MHC. Parallel with the increase in the number of lymphocytes, larger numbers of small and large muscle fibres expressed high levels of either donor, host or both, class I — but not class II — molecules. Surprisingly, immune reactions continued over several months, causing gradual loss of muscle tissue. Donor class I molecules persisted for more than 6 months after Cyclosporin A withdrawal, clearly indicating survival of donor muscle fibres despite ongoing rejection. Indirect evidence on the other hand suggests additional loss of host fibres, possibly caused by cytokine release from the immune cells (bystander damage). We conclude that transient treatment with Cyclosporin A induced a kind of tolerance related to the maturation and down-regulation of class I antigens in donor muscle fibres. It is suggested that the start of immune reaction following Cyclosporin A withdrawal is initiated by remaining small amounts of donor MHC molecules, possibly related to the continuous proliferation of the cell-lined-derived donor myoblasts.
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References
Appleyard, S. T., Dunn, M. J., Dubowitz, V. &Rose, M. L. (1985) Increased expression of HLA ABC class I antigens by muscle fibres in Duchenne muscular dystrophy, inflammatory myopathy, and other neuromuscular disorders.Lancet 361–3 (Feb 16).
Arahata, K. &Engel, A. G. (1988) Monoclonal antibody analysis of mononuclear cells in myopathies. IV: Cell-mediated cytotoxicity and muscle fiber necrosis.Annals of Neurology 23, 168–73.
Bao, S., King, N. J. C. &Dos Remedios, C. G. (1990) Elevated MHC class I and II antigens in cultured human embryonic myoblasts following stimulation with gamma-interferon.Immunology and Cell Biology 68, 235–42.
Baroffio, A., Aubry, J. P., Kaelin, A., Krause, R. M., Hamann, M. &Bader, C. R. (1993) Purification of human muscle satellite cells by flow cytometry.Muscle and Nerve 16, 498–505.
Beauchamp, J. R., Abraham, D. J., Bou-Gharios, G., Partridge, T. A. &Olsen, K. J. (1992) Expression and function of heterotypic adhesion molecules during differentiation of human skeletal muscle in culture.American Journal of Pathology 140, 387–401.
Bornemann, A. &Schmalbruch, H. (1992) Desmin and vimentin in regenerating muscles.Muscle and Nerve 15, 14–20.
Dhawan, J., Pan, L. C., Pavlath, G. K., Travis, M. A., Lanctot, A. M. &Blau, H. M. (1991) Systemic delivery of human growth hormone by injection of genetically engineered myoblasts.Science 254, 1509–12.
Dialynas, D. P., Quan, Z. S., Wall, K. A., Pierres, A., Quintans, J., Loken, M. R., Pierres, M. &Fitch, F. W. (1983) Characterization of the murine T cell surface molecule, designated L3T4, identified by monoclonal antibody GK 1.5: similarity of L3T4 to the human Leu-3/T4 molecule.Journal of Immunology 13, 2445–51.
Flores, E. A., Bistrian, B. R., Pomposelli, J. J., Dinarello, C. A., Blackburn, G. L. &Istfan, N. W. (1989) Infusion of tumor necrosis factor/cachetin promotes muscle catabolism in the rat.Journal of Clinical Investigation 83, 1614–22.
Goebels, N., Michaelis, D., Wekerle, H. &Hohlfeld, R. (1992) Human myoblasts as antigen-presenting cells.Journal of Immunology 149, 661–7.
Goodman, M. N. (1991) Tumor necrosis factor induces skeletal muscle protein breakdown in rats.American Journal of Physiology 260, E727–30.
Green, C. J. (1975) Neuroleptanalgesic drug combinations in the anaesthetic management of small laboratory animals.Laboratory Animals 9, 161–78.
Harrison, S. A., Mondino, B. J. &Kagan, J. M. (1990) Modulation of HLA antigen expression on conjunctival fibroblasts by gamma-interferon.Investigative Ophthalmology and Visual Science 31, 163–7.
Hoffman, E. P. (1993) Myoblast transplantation: what's going on?Cell Transplantation 2, 49–57.
Hohlfeld, R. &Engel, A. G. (1990) Induction of HLA-DR expression on human myoblasts with interferon-gamma.American Journal of Pathology 136, 503–8.
Hohlfeld, R. &Engel, A. G. (1994) The immunobiology of muscle.Immunology Today 15, 269–74.
Huard, J., Bouchard, J. P., Roy, R., Labrecque, C., Dansereau, G., Lemieux, B. &Tremblay, J. P. (1991) Myoblast transplantation produced dystrophin-positive muscle fibres in a 16-year-old patient with Duchenne muscular dystrophy.Clinical Science 81, 287–8.
Huard, J., Roy, R., Bouchard, J. P., Malouin, F., Richards, C. L. &Tremblay, J. P. (1992) Human myoblast transplantations between immunohistocompatible donors and recepients produce immune reactions.Transplantation Proceedings 24, 3049–51.
Irintchev, A. &Wernig, A. (1987) Muscle damage and repair in voluntarily running mice: strain and muscle differences.Cell and Tissue Research 249, 509–21.
Irintchev, A., Draguhn, A. &Wernig, A. (1990) Reinnervation and recovery of mouse soleus muscle after long-term denervation.Neuroscience 39, 231–43.
Irintchev, A., Zeschnigk, M., Starzinski-Powitz, A. &Wernig, A. (1994) Expression pattern of M-cadherin in normal, denervated and regenerating mouse muscles.Developmental Dynamics 199, 326–37.
Jiao, S., Gurevich, V. &Wolff, J. A. (1993) Long-term correction of rat model of Parkinson's disease by gene therapy.Nature 362, 450–3.
Karpati, G. (1990) Immunological aspects of histoincompatible myoblast transfer into non-tolerant hosts. InMyoblast Transfer Therapy (edited byGriggs, R. C. &Karpati, G.) p. 31–4. New York: Plenum Press.
Karpati, G., Pouilot, Y. &Carpenter, S. (1988) Expression of immunoreactive major histocompatibility complex products in human skeletal muscles.Annals of Neurology 23, 64–72.
King, N. J. C., Sinickas, V. G. &Blanden, R. V. (1986) H-2D and H-2K antigens are independently regulated in mouse embryo fibroblasts.Experimental and Clinical Immunogenetics 2, 206–14.
Klaus, G. G. B. &Kunkl, A. (1983) Effects of cyclosporine on the immune system of the mouse. II. Cyclosporine inhibits the effector function of primary T helper cells, but not helper cell priming.Transplantation 36, 80–4.
Koch, S., Koch, N., Robinson, P. &Hämmerling, G. (1993) Comparison of allogeneic and xenogeneic determinants on the H-2Kk molecule.Transplantation 36, 177–80.
Ledbetter, J. A. &Herzenberg, J. A. (1970) Xenogeneic monoclonal antibodies to mouse lymphoid differentiation antigens.Immunology Reviews 47, 63–90.
Mantegazza, R., Hughes, S. M., Mitchell, D., Travis, M. A., Blau, H. M. &Steinman, L. (1991) Modulation of MHC class II antigen expression in human myoblasts after treatment with IFN-gamma.Neurology 41, 1128–32.
Michaelis, D., Goebels, N. &Hohlfeld, R. (1993) Constitutive and cytokine-induced expression of human leukocyte antigens and cell adhesion molecules by human myotubes.American Journal of Pathology 143, 1142–9.
Milton, A. D. &Fabre, J. W. (1985) Massive induction of donor-type class I and class II major histocompatibility complex antigens in rejecting cardiac allografts in the rat.Journal of Experimental Medicine 161, 98–112.
Momburg, F., Koch, N., Möller, P., Moldenhauer, G., Butcher, G. W. &Hämmerling, G. J. (1986a) Differential expression of Ia and Ia-associated invariant chain in mouse tissues afterin vivo treatment with IFN-gamma.Journal of Immunology 136, 940–8.
Momburg, F., Koch, N., Möller, P., Moldenhauer, G. &Hämmerling, G. J. (1986b)In vivo induction of H-2K/D antigens by recombinant interferon-gamma.European Journal of Immunology 16, 551–7.
Morgan, J. E. &Partridge, T. A. (1992) Cell transplantation and gene therapy in muscular dystrophy.BioEssays 14, 641–5.
Nathan, C. F. (1987) Secretory products of macrophages.Journal of Clinical Investigation 79, 319–26.
Orimo, S., Hiyamuta, E., Arahata, K. &Sugita, H. (1991) Analysis of inflammatory cells and complement C3 in bupivacaine-induced myonecrosis.Muscle and Nerve 14, 515–20.
Ozato, K., Mayer, N. M. &Sachs, D. H. (1982) Monoclonal antibodies to mouse major histocompatibility complex antigens. IV. A series of hybridoma clones producing anti-H-2d antibodies and an examination of expression of H-2d antigens on the surface of these cells.Transplantation 34, 113–20.
Partridge, T. A. (1991) Myoblast transfer: a possible therapy for inherited myopathies?Muscle and Nerve 14, 197–212.
Ponder, B. A. J., Wilkinson, M. M., Wood, M. &Westwood, J. H. (1983) Immunohistochemical demonstration of H2 antigens in mouse tissue sections.Journal of Histochemistry and Cytochemistry 31, 911–19.
Rando, T. A. &Blau, H. M. (1994) Primary mouse myoblast purification, characterization, and transplantation for cell-mediated gene therapy.Journal of Cell Biology 125, 1275–87.
Round, J. M., Jones, D. A. &Cambridge, G. (1987) Cellular infiltrates in human skeletal muscle: exercise induced damage as a model for inflammatory muscle disease?Journal of the Neurological Sciences 82, 1–11.
Roy, R., Dansereau, G., Tremblay, J. P., Belles-Isles, M., Huard, J., Labrecque, C. &Bouchard, J. P. (1991) Expression of major histocompatibility complex antigens on human myoblasts.Transplantation Proceedings 23, 799–801.
Sandvig, S., Laskay, T., Andersson, J., De Lay, M. &Andersson, U. (1987) Gamma-interferon is produced by CD3+ and CD3− lymphocytes.Immunology Reviews 97, 51–65.
Sofroniew, M. V. &Schrell, U. (1982) Long-term storage and regular repeated use of diluted antisera in glass staining jars for increased sensitivity, reproducibility, and convenience of single- and two-color light microscopic immunocytochemistry.Journal of Histochemistry and Cytochemistry 30, 504–11.
Unanue, E. R. &Allen, P. M. (1987) The basis for the immunoregulatory role of macrophages and other accessory cells.Science 236, 551–7.
Wang, Y. C., Herskowitz, A., Gu, L. B., Kanter, K., Lattouf, O., Sell, K. W. &Ahmed-Ansari, A. (1991) Influence of cytokines and immunosuppressive drugs on major histocompatibility complex class I/II expression by human cardiac myocytesin vitro.Human Immunology 31, 123–33.
Warrens, A. N., Zhang, J. Y., Sidhu, S., Watt, D. J., Lombardi, J., Sewry, C. A. &Lechler, R. I. (1994) Myoblasts fail to stimulate T cells but induce tolerance.International Immunology 6, 847–53.
Watt, D. J., Morgan, J. E. &Partridge, T. A. (1991) Allografts of muscle precursor cells persist in the nontolerized host.Neuromuscular Disorders 1, 345–55.
Wernig, A. &Irintchev, A. (1995) ‘Bystander’ damage of host muscle caused by implantation of MHC-compatible myogenic cells.Journal of the Neurological Sciences, in press.
Wernig, A., Irntchev, A., Härtling, A., Stephan, G., Zimmermann, K. &Starzinski-Powitz, A. (1991) Formation of new muscle fibres and tumours after injection of cultured myogenic cells.Journal of Neurocytology 20, 982–97.
Wernig, A., Irintchev, A. &Lange, G. (1995) Functional effects of myoblast implantation into histoincompatible mice with or without immunosuppression.Journal of Physiology, in press.
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Irintchev, A., Zweyer, M. & Wernig, A. Cellular and molecular reactions in mouse muscles after myoblast implantation. J Neurocytol 24, 319–331 (1995). https://doi.org/10.1007/BF01186543
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DOI: https://doi.org/10.1007/BF01186543