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Antigen Presenting Cells: Detection and Quantification of a Cytochrome c Determinant Important for Activation of T-Cells on Bone Marrow Derived Macrophages by using Specific Anti Cytochrome c Monoclonal Antibody

  • Stéphane Demotz
  • Claudio Vita
  • Giampietro Corradin
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 225)

Summary

The region of the horse cytochrome c molecule recognized by flab SJL2-4 specific for the denatured form of the protein was located around residues 22–28. Binding studies on antigen pulsed macrophages were also performed.Surprisingly, heme peptide 1–65 was not recognised by Mab when bound on macrophages. This correlates with the incapacity of the same peptide to activate the T-cell clone 2–16. Binding sites on antigen pulsed macrophages varied between 0.5−2×106 per cell depending on the conditions used. The expression of the antigenic determinant as detected by Mab was also followed under different conditions (chloroquine, trypsin treatment) and time. Kinetics parameters of the antigen-antibody reaction in solution and on antigen bound macrophages were also determined and are dramatically different. This is correlated with a different structure of the peptide in solution and on macrophage cell surface.

Keywords

Association Constant Antigenic Peptide Antigenic Site Scatchard Analysis Candida Krusei 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Allen, P. M., Beller, D. I., Braun, J., and Unanue, E. R., 1984, The handling of Listeria monocytogenes by macrophages: The search for an immunogenic molecule in antigen presentation, J. Immunol., 132: 323–331.PubMedGoogle Scholar
  2. Atherton, E., Logan, C. J., and Sheppard, R. C., 1979, Peptide synthesis. Part 2. Procedure for solid phase synthesis using N -fluorenylmethoxycarbonylamino acids on polyamide supports. Synthesis of substance P and of acyl carrier 65–74 decapeptide, Biorganic Chem., 8: 351–370.Google Scholar
  3. Babbitt, B. P., Allen, P. M., Matsueda, G., Haber, E., and Unanue, E. R., 1985, Binding of immunogenic peptides to la histocompatibility molecules. Nature, 317: 359–361.PubMedCrossRefGoogle Scholar
  4. Babbitt, B. P., Matsueda, G., Haber, E., Unanue, R. R., and Allen, P. M., 1985, Antigenic competition at the level of peptide-Ia binding. Proc. Natl. Acad. Sci. USA, 83: 4509–4513.CrossRefGoogle Scholar
  5. Buus, S., Colon, S., Smith, C., Freed, J. H.., Miles, C., and Grey, H. M., 1986, Interaction between a “processes” ovalbumin peptide and la molecules, Proc. Natl. Acad. Sci. USA, 83: 3968–3971.PubMedCrossRefGoogle Scholar
  6. Chesnut, R. W., Colon, S. M., and Grey, H. M., 1982, Requirements for the processing of antigens by antigen-presenting B cells. I. Functional comparison of B cell tumors and macrophages, J. Immunol., 129: 2382–2388.PubMedGoogle Scholar
  7. Corradin, G., and Harbury, H. A., 1970, Cleavage of cytochrome c with cyanogen bromide. Biochim. Biophys. Acta, 221: 489–496.PubMedCrossRefGoogle Scholar
  8. Corradin, G., Etlinger, E. H., and Chiller, J. M., 1977, Lymphocyte specificity to protein antigens. I. Characterization of the antigen-induced in vitro T cell-dependent proliferative response with lymph node cells from primed mice. J. Immunol., 119: 1048–1053.PubMedGoogle Scholar
  9. Corradin, G., Juillerat, M. A., Vita, C., and Engers, H. D., 1983, Fine specificity of a Balb/c T cell clone directed against beef apocytochrome c. Mol. Immunol., 20: 763–768.PubMedCrossRefGoogle Scholar
  10. Corradin, G., and Engers, H. D., 1984. Inhibition of antigen induced T-cell clone proliferation by antigen-specific antibodies. Nature, 308: 547–548.PubMedCrossRefGoogle Scholar
  11. Corradin, G., Juillerat, M. A., and Engers, H. D., 1984, Differential effects of two anti-apo-cytochrome c-specific monoclonal antibodies on the function of apo-cytochrome c-specific murine T cell clones. J. Immunol., 133: 2915–2919.PubMedGoogle Scholar
  12. Corradin, G. P., Wallace, C. J. A., Proudfoot, A. E. I., and Baumhüter, S., 1987, Murine T cell response specific for cytochrome c, “CRC”, Ed. J.A. Berzofsky and E. Sercarz, (in press).Google Scholar
  13. De Lisi, C., and Berzofsky, J. A., 1985, T cell antigenic site tend to be amphipathic structures, Proc. Natl. Acad. Sci. USA, 82: 7048–7052.CrossRefGoogle Scholar
  14. Demotz, S., Mozzarelli, A., Baumhüter, S., and Corradin, G., 1984, Biochemical aspects of antigen degradation and presentation by macrophages, in Abstracts “6th European Immunology Meeting”, Interlaken, pp. 370.Google Scholar
  15. Falo, L. D., Benacerraf, Jr. B., and Rock, K. L., 1986, Phospholipase treatment of accessory cells that have been exposed to antigen selectively inhibits antigen-specific Ia-restricted, but not allo-specific, stimulation of T lymphocytes. Proc. Natl. Sci. USA, 83: 6994–6997.CrossRefGoogle Scholar
  16. Fanger, M. W., and Harbury, H. A., 1965, Trifluoroacetylated cytochrome c. Biochemistry, 4: 2541–2545.CrossRefGoogle Scholar
  17. Fisher, W. R. H., Taniuchi, H., and Anfinsen, C. B., 1973, On the role of the heure in the formation of the structure of cytochrome c. J. Biol. Chem., 248: 3188–3195.PubMedGoogle Scholar
  18. Harbury, H. A., and Loach, P. A., 1960, Oxidation-linked proton functions in heure octa-and undecapeptides from mammalian cytochrome c, J. Biol. Chem., 2345: 3640–3645.Google Scholar
  19. Karush, F., 1978, Immunoglobulins, “Comprehensive Immunology”, G. Litman and R.A. Good, eds, Plenum Publishing Corp. New York, Vol. V, pp 85–116.Google Scholar
  20. Koehler, G., and Milstein, C., 1975, Continous cultures of fused cells secreting antibodies of predefined specificity. Nature, 256: 495–497.CrossRefGoogle Scholar
  21. Margoliash, E., and Wasalek, 0. F., 1967, Cytochrome c from vertebrate and invertebrate sources, in: “Methods in Enzymology”, R. W. Estabrook and M. E. Pullman, eds, Academic Press, Inc., New York, Vol. 10, p. 339.Google Scholar
  22. Meerpohl, H.-G., Lohmann-Matthes, M.-L., and Fischer, H., 1976, Studies on the activation of mouse bone marrow derived macrophages by the macrophage cytotoxicity factor, Eur. J. Immunol., 6: 213–217.PubMedCrossRefGoogle Scholar
  23. Parikh, I., March, S., Cuatrecasas, P., 1974, “Methods in enzymology”, W. B., Jakoby and M. Wilchek, eds, Academic Press, New York, Vol. 34, pp 77–102.Google Scholar
  24. Pincus, M. R., Gerewitz, F., Schwartz, R. H., and Sheraga, H. A., 1983, Correlation between the conformation of cytochrome c peptides and their stimulatory activity in a T-lymphocyte prolifération assay, Proc. Natl. Acad. Sci. USA, 80: 3297–3300.PubMedCrossRefGoogle Scholar
  25. Sela, M., 1969, Antigenicity: Some molecular aspects, Science, 166: 1365–1374.PubMedCrossRefGoogle Scholar
  26. Shimonkevitz, R., Kappler, J., Marrack, P., and Grey, H., 1983, Antigen recognition by H-2-restricted T cells. I. Cell-free antigen processing. J. Exp. Med., 158: 303–316.PubMedCrossRefGoogle Scholar
  27. Shimonkevitz, R., Colon, S., Kappier, J. W., Marrack, P., Grey, H. M., 1984, Antigen recognition by H-2 restricted T cell. II. A tryptic ovalbumin peptide that substitues for processed antigen. J. Immunol., 4: 2067–2074.Google Scholar
  28. Wu, C. S. C., Ikeda, K., and Yang, J. T., 1981, Ordered conformation of polypeptides and proteins in acidic dodecyl sulfate solution, Biochemistry, 20: 566–570.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Stéphane Demotz
    • 1
  • Claudio Vita
    • 2
  • Giampietro Corradin
    • 1
  1. 1.Institut de BiochimieUniversité de LausanneEpalingesSwitzerland
  2. 2.Istituto di Chimica OrganicaUniversita di PadovaPadovaItaly

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