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Generation and characterization of mouse monoclonal antibodies to the myelin-associated glycoprotein (MAG)

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Abstract

A panel of mouse monoclonal antibodies to rat and human myelin-associated glycoprotein (MAG) was developed. Normal mice were unresponsive to rat MAG, and successful immunization with rat MAG was only achieved in autoimmune NZB mice. By contrast, all strains of mice were responsive to human MAG. The monoclonal antibodies developed differ with respect to immunoglobulin type, their specificity for human and/or rat MAG, and their recognition of protein or carbohydrate epitopes in MAG. In general, the antibodies that react with the protein backbone recognize both rat and human MAG, whereas a large number of the monoclonal antibodies recognize a carbohydrate determinant in human MAG that is not in rat MAG. Immunocytochemical staining of adult human spinal cord with the monoclonal antibodies resulted in periaxonal staining of myelin sheaths, similar to that produced by well-defined, rabbit, polyclonal anti-MAG serum. In addition, the antibodies recognizing, carbohydrate determinants in human MAG strongly stained oligodendrocyte cytoplasm. These monoclonal antibodies will be of value for the further chemical and biological characterization of MAG.

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References

  1. Quarles, R. H., Everly, J. L., andBrady, R. O. 1973. Evidence for the close association of a glycoprotein with myelin in rat brain. J. Neurochem. 21:1177–1191.

    PubMed  Google Scholar 

  2. Figlewicz, D. A., Quarles, R. H., Johnson, D., Barbarash G. R., andSternberger, N. H. 1981. Biochemical demonstration of the myelin-associated glycoprotein in the peripheral nervous system. J. Neurochem. 37:749–758.

    PubMed  Google Scholar 

  3. Sternberger, N. H., Quarles, R. H., Itoyama, Y. andWebster H. DeF. 1979. Myelin-associated glycoprotein demonstrated immunocytochemically in myelin and myelin-forming cells of developing rat. Proc. Natl. Acad. Sci., USA 76:1510–1514.

    Google Scholar 

  4. Trapp, D. B., andQuarles, R. H. 1982. Presence of myelin-associated glycoprotein correlates with alterations in the periodicity of peripheral myelin. J. Cell Biol. 92:877–882.

    PubMed  Google Scholar 

  5. Dobersen, M. J., MacIntosh, T. D., Quarles, R. H., andBrady, R. O. 1983. Antibody 7E10: A monoclonal antibody to myelin associated glycoprotein (MAG). J. Neurochem. 41:S59A (abstr.)

    Google Scholar 

  6. Quarles, R. H., andPasnak, C. L. 1977. A rapid procedure for selectively isolating the major glycoprotein from purified rat brain myelin. Biochem. J. 163:635–637.

    PubMed  Google Scholar 

  7. Quarles, R. H., Barbarash, G. R., Figlewicz, D. A., andMcIntyre, L. J. 1983. Biochim. Biophys. Acta 757:140–143.

    PubMed  Google Scholar 

  8. Prabhakar, B. S., Haspel, M. V. andNotkins, A. 1984. Monoclonal antibody techniques applied to viruses.in C. Maramorosch andH. Koprowski (eds.), Methods in Virology, Academic Press, New York, in press.

    Google Scholar 

  9. Galfre, G., andMilstein, C. 1981. Preparation of monoclonal antibodies: Strategies and procedures. Meth. Enzymol. 73:3–46.

    PubMed  Google Scholar 

  10. Johnson, D., Quarles, R. H., andBrady, R. O. 1982. A radioimmunoassay for the myelin-associated glycoprotein. J. Neurochem. 39:1356–1362.

    PubMed  Google Scholar 

  11. Edge, A. S. B., Faltynek, C. R., Hof, L., Reichert, L. E., andWeber, P. 1981. Deglycosylation of glycoproteins by trifluoromethanesulfonic acid. Anal. Biochem. 118:131–137.

    PubMed  Google Scholar 

  12. Elder, J. H., andAlexander, S. 1982. endo-B-N-Acetylglucosaminidase F: Endoglycosidase fromFlavobacterium menningosepticum that cleaves both high mannose and complex glycoproteins. Proc. Natl. Acad. Sci. USA 79:4540–4544.

    PubMed  Google Scholar 

  13. Laemmli, V. K., andFavre, M. 1973. Maturation of the head of bacteriophage T4. 1. DNA packing events. J. Mol. Biol. 80:575–600.

    PubMed  Google Scholar 

  14. Towbin, H., Staehelin, T., andGordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci., USA 76:4350–4354.

    Google Scholar 

  15. Quarles, R. H., Johnson, D., Brady, R. O., andSternberger, N. H. 1981. Preparation and characterization of antisera to the myelin-associated glycoprotein. Neurochem. Res. 6:1115–1127.

    PubMed  Google Scholar 

  16. Ilyas, A. A., Quarles, R. H., MacIntosh T. D., Dobersen, M. J., Trapp, B. D., Dalakas, M. C., andBrady, R. O. 1984. IgM in a human neuropathy related to paraproteinemia binds to a carbohydrate determinant in the myelin-associated glycoprotein and to a ganglioside. Proc. Natl. Acad. Sci., USA 81:1225–1229.

    Google Scholar 

  17. Itoyama, Y., Sternberger, N. H., Webster, H. DeF., Quarles, R. H., Cohen, S. R., andRichardson, E. P. Jr. 1980. Immunocytochemical observations on the distribution of myelin-associated glycoprotein and myelin basic protein in multiple sclerosis lesions. Ann Neurol. 7:167–177.

    PubMed  Google Scholar 

  18. Howie, J. B., andHelyer, B. J. 1968. The immunology and pathology of NZB mice. Adv. Immunol. 9:215–266.

    PubMed  Google Scholar 

  19. Sato, S., Quarles, R. H., andBrady, R. O. 1982. Susceptibility of the myelin-associated glycoprotein and basic protein to a neutral protease in highly purified myelin from human and rat brain. J. Neurochem. 39:97–105.

    PubMed  Google Scholar 

  20. Inuzuka, T., Quarles, R. H., Noronha, A. B., Dobersen, M. J., andBrady, R. O., 1984. A human lymphocyte antigen is shared with a group of glycoproteins in peripheral nerve, Neurosci. Let., 51:105–111.

    Google Scholar 

  21. Nobile-Orazio, E., Hays, A. P., Latov, N., Perman, G., Golier, J., Shy, M., andFreddo, L. 1984. Specificity of mouse and human monoclonal antibodies to the myelin-associated glycoprotein (MAG). Neurology 34:1336–1342.

    PubMed  Google Scholar 

  22. Abo, T., andBalch, C. M. 1981. A differentiation antigen of human NK and K cells identified by a monoclonal antibody (HNK-1). J. Immunol. 127:1024–1029.

    PubMed  Google Scholar 

  23. Tilden, A. B., Abo, T., andBalch, C. M. 1983. Suppressor cell function of human granular lymphocytes identified by the HNK-1 (Leu 7) monoclonal antibody. J. Immunol. 130:11781–11785.

    Google Scholar 

  24. McGarry, R. C., Helfand, S. L., Quarles, R. H., andRoder, J. C. 1983. Recognition of myelin-associated glycoprotein by the monoclonal antibody HNK-1. Nature 306:376–378.

    PubMed  Google Scholar 

  25. Sato, S., Baba, H., Tanaka, M., Yanagisawa, K., andMiyatake, Y. 1983. Antigenic determinant shared between myelin-associated glycoprotein from human brain and natural killer cells. Biomed. Res. 4:489–494.

    Google Scholar 

  26. Murray, N., andSteck, A. J. 1984. Indication of a possible role in a demyelinating neuropathy for an antigen shared between myelin and NK cells. Lancet 1(8379):711–713.

    PubMed  Google Scholar 

  27. Ilyas, A., Quarles, R. H., andBrady, R. O. 1984. The monoclonal antibody HNK-1 reacts with a human peripheral nerve ganglioside. Biochem. Biophys. Res. Commun. 122:1206–1211.

    PubMed  Google Scholar 

  28. Dobersen, M. J., Gascon, P., Trost, S., Hammer, J. A., Goodman, S., Noronha, A. B., O'Shannessy, D. J., Brady, R. O., andQuarles, R. H., 1985. Murine monoclonal antibodies to the myelin-associated glycoprotein react with large granular lymphocytes of human blood. Proc. Natl. Acad. Sci. USA, 82:552–555.

    PubMed  Google Scholar 

  29. Liebowitz, S., Gregson, N. A., Kennedy, M., andKahn, S. N. 1983. IgM paraproteins with immunological specificity for a Schwann cell component and peripheral nerve myelin in patients with polyneuropathy. J. Neurol. Sci. 59:153–165.

    PubMed  Google Scholar 

  30. Dellagi, K., Depouey, P., Brouet, J. C., Billecocq, A., Gomez, D., Clauvel, J. P., andSeligmann, M. 1983. Waldenstrom's macroglobulinemia and peripheral neuropathy: a clinical and immunological study of 25 patients. Blood 62:280–285.

    PubMed  Google Scholar 

  31. Abrams, G. M., Latov, N., Hays, A. P., Sherman, W., andZimmerman, E. A. 1982. Immunocytochemical studies of human peripheral nerve with serum from patients with polyneuropathy and paraproteinemia. Neurology 32:821–826.

    PubMed  Google Scholar 

  32. Schober, R., Itoyama, Y., Sternberger, N. H., Trapp, B. D., Richardson, E. P., Asbury, A. K., Quarles, R. H., andWebster, H. DeF. 1981. Immunocytochemical study of PO glycoprotein, P1 and P2 basic proteins, and myelin-associated glycoprotein (MAG) in lesions of idiopathic polyneuritis. Neuropath. Applied Neurobiol. 7:437–451.

    Google Scholar 

  33. Webster, H. DeF., Palkovits, C. G., Stoner, G. L., Favilla, J. T., Frail, D. E., andBraun P. E. 1983. Myelin associated glycoprotein-Electron microscopic immunocytochemical localization in compact developing and adult central nervous system myelin. J. Neurochem. 41:1469–1479.

    PubMed  Google Scholar 

  34. Favilla, J. T., Frail, D. E., Palkovitz, C. G., Stoner, G. L., Braun, P. E., andWebster, H. DeF. 1984. Myelin-associated glycoprotein, distribution in human central nervous tissue studied immunocytochemically with monoclonal antibody. J. Neuroimmunol. 6:19–30.

    PubMed  Google Scholar 

  35. Trapp, B. D., andQuarles, R. H. 1984. Immunocytochemical localization of the myelin-associated glycoprotein: Fact or artifact? J. Neuroimmunol. 6:231–249.

    PubMed  Google Scholar 

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Author notes

  1. Michael J. Dobersen

    Present address: Barbara Davis Center for Childhood Diabetes, University of Colorado Medical Center, 4200 East 9th Avenue, Box # B-140, 80262, Denver, Colorado

Authors and Affiliations

  1. Developmental and Metabolic Neurology Branch and Laboratory of Molecular Genetics NINCDS, NIH, Park Bldg., Rm. 425, 20205, Bethesda, Maryland

    Michael J. Dobersen, Jeffrey A. Hammer, Antonio B. Noronha, Tracy D. MacIntosh, Bruce D. Trapp, Roscoe O. Brady &  Richard H. Quarles

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  1. Michael J. Dobersen
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Dobersen, M.J., Hammer, J.A., Noronha, A.B. et al. Generation and characterization of mouse monoclonal antibodies to the myelin-associated glycoprotein (MAG). Neurochem Res 10, 499–513 (1985). https://doi.org/10.1007/BF00964654

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