Abstract
The aim of this study was to investigate the susceptibility of human myotubes to lysis by the two major types of cytotoxic effector cells, CD3+CD8+ cytotoxic T cells (CTL) and CD16+CD56+ natural killer (NK) cells. The myoblasts preparations used as target cells were greater than 90% pure as assessed by immunostaining with the Leu19 monoclonal antibody (MAb) that cross-reacts with the neural cell adhesion molecule N-CAM. Allospecific CTL lines were generated from mixed lymphocyte cultures, and freshly isolated allogeneic and autologous peripheral blood cells were used as a source of NK cells. The cytotoxicity was observed under phase optics and by immunoelectron microscopy, and was quantitated with a chromium release assay. Myotubes were efficiently killed by allospecific CTL and by autologous and allogeneic NK cells. The killing by CTL was inhibited with an anti-class I HLA MAb, and the killing by NK cells was inhibited by depleting peripheral blood cells of CD16+ cells with anti-CD16 MAb and complement. The results have important implications for myoblast transplantation, an experimental therapy of muscular dystrophy.
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Selected References
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- Arahata K., Engel A. G. Monoclonal antibody analysis of mononuclear cells in myopathies. III: Immunoelectron microscopy aspects of cell-mediated muscle fiber injury. Ann Neurol. 1986 Feb;19(2):112–125. doi: 10.1002/ana.410190203. [DOI] [PubMed] [Google Scholar]
- Blau H. M., Webster C. Isolation and characterization of human muscle cells. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5623–5627. doi: 10.1073/pnas.78.9.5623. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chervenak R., Wolcott R. M. Target cell expression of MHC antigens is not (always) a turn-off signal to natural killer cells. J Immunol. 1988 Jun 1;140(11):3712–3716. [PubMed] [Google Scholar]
- Edelman G. M. CAMs and Igs: cell adhesion and the evolutionary origins of immunity. Immunol Rev. 1987 Dec;100:11–45. doi: 10.1111/j.1600-065x.1987.tb00526.x. [DOI] [PubMed] [Google Scholar]
- Gabridge M. G., Dougherty E. P. Improved method for transmission electron microscopy of ciliated cell monolayers maintained on gas-permeable membranes. J Microsc. 1983 Nov;132(Pt 2):165–169. doi: 10.1111/j.1365-2818.1983.tb04268.x. [DOI] [PubMed] [Google Scholar]
- Gutmann D. H., Fischbeck K. H. Molecular biology of Duchenne and Becker's muscular dystrophy: clinical applications. Ann Neurol. 1989 Aug;26(2):189–194. doi: 10.1002/ana.410260202. [DOI] [PubMed] [Google Scholar]
- Hoffman E. P., Brown R. H., Jr, Kunkel L. M. Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell. 1987 Dec 24;51(6):919–928. doi: 10.1016/0092-8674(87)90579-4. [DOI] [PubMed] [Google Scholar]
- Hohlfeld R., Engel A. G. Induction of HLA-DR expression on human myoblasts with interferon-gamma. Am J Pathol. 1990 Mar;136(3):503–508. [PMC free article] [PubMed] [Google Scholar]
- Karpati G., Pouliot Y., Carpenter S. Expression of immunoreactive major histocompatibility complex products in human skeletal muscles. Ann Neurol. 1988 Jan;23(1):64–72. doi: 10.1002/ana.410230111. [DOI] [PubMed] [Google Scholar]
- Karpati G., Pouliot Y., Zubrzycka-Gaarn E., Carpenter S., Ray P. N., Worton R. G., Holland P. Dystrophin is expressed in mdx skeletal muscle fibers after normal myoblast implantation. Am J Pathol. 1989 Jul;135(1):27–32. [PMC free article] [PubMed] [Google Scholar]
- Kärre K., Ljunggren H. G., Piontek G., Kiessling R. Selective rejection of H-2-deficient lymphoma variants suggests alternative immune defence strategy. Nature. 1986 Feb 20;319(6055):675–678. doi: 10.1038/319675a0. [DOI] [PubMed] [Google Scholar]
- Lanier L. L., Le A. M., Civin C. I., Loken M. R., Phillips J. H. The relationship of CD16 (Leu-11) and Leu-19 (NKH-1) antigen expression on human peripheral blood NK cells and cytotoxic T lymphocytes. J Immunol. 1986 Jun 15;136(12):4480–4486. [PubMed] [Google Scholar]
- Lanier L. L., Testi R., Bindl J., Phillips J. H. Identity of Leu-19 (CD56) leukocyte differentiation antigen and neural cell adhesion molecule. J Exp Med. 1989 Jun 1;169(6):2233–2238. doi: 10.1084/jem.169.6.2233. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Leiden J. M., Karpinski B. A., Gottschalk L., Kornbluth J. Susceptibility to natural killer cell-mediated cytolysis is independent of the level of target cell class I HLA expression. J Immunol. 1989 Mar 15;142(6):2140–2147. [PubMed] [Google Scholar]
- Main E. K., Monos D. S., Lampson L. A. IFN-treated neuroblastoma cell lines remain resistant to T cell-mediated allo-killing, and susceptible to non-MHC-restricted cytotoxicity. J Immunol. 1988 Nov 1;141(9):2943–2950. [PubMed] [Google Scholar]
- Mandel J. L. Dystrophin. The gene and its product. Nature. 1989 Jun 22;339(6226):584–586. doi: 10.1038/339584a0. [DOI] [PubMed] [Google Scholar]
- Moretta A., Pantaleo G., Moretta L., Cerottini J. C., Mingari M. C. Direct demonstration of the clonogenic potential of every human peripheral blood T cell. Clonal analysis of HLA-DR expression and cytolytic activity. J Exp Med. 1983 Feb 1;157(2):743–754. doi: 10.1084/jem.157.2.743. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nitta T., Yagita H., Sato K., Okumura K. Involvement of CD56 (NKH-1/Leu-19 antigen) as an adhesion molecule in natural killer-target cell interaction. J Exp Med. 1989 Nov 1;170(5):1757–1761. doi: 10.1084/jem.170.5.1757. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Partridge T. A., Morgan J. E., Coulton G. R., Hoffman E. P., Kunkel L. M. Conversion of mdx myofibres from dystrophin-negative to -positive by injection of normal myoblasts. Nature. 1989 Jan 12;337(6203):176–179. doi: 10.1038/337176a0. [DOI] [PubMed] [Google Scholar]
- Royer H. D., Reinherz E. L. T lymphocytes: ontogeny, function, and relevance to clinical disorders. N Engl J Med. 1987 Oct 29;317(18):1136–1142. doi: 10.1056/NEJM198710293171807. [DOI] [PubMed] [Google Scholar]
- Schubert W., Zimmermann K., Cramer M., Starzinski-Powitz A. Lymphocyte antigen Leu-19 as a molecular marker of regeneration in human skeletal muscle. Proc Natl Acad Sci U S A. 1989 Jan;86(1):307–311. doi: 10.1073/pnas.86.1.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Trinchieri G. Biology of natural killer cells. Adv Immunol. 1989;47:187–376. doi: 10.1016/S0065-2776(08)60664-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zuckermann F. A., Head J. R. Murine trophoblast resists cell-mediated lysis. I. Resistance to allospecific cytotoxic T lymphocytes. J Immunol. 1987 Nov 1;139(9):2856–2864. [PubMed] [Google Scholar]