Advertisement

Preparation and Characterization of Antisera and of Murine Monoclonal Antibodies to Human Glioma-Associated Antigen(s)

  • Masafumi Matsuda
  • Duncan K. Fischer
  • Raj K. Narayan
  • M. Zouhair Atassi
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 303)

Abstract

Human glioma-associated markers can be exploited for the development of new diagnostic strategies and treatment modalities for these malignancies. A goat antiserum was first raised against human anaplastic astrocytoma (AC or AA) and glioblastoma multiforme (GB or GBM) extracts. Extensive sequential absorptions with normal brain tissue, normal serum, and human serum albumin (HSA) gave an antibody fraction specific for glioma. Balb/c mice were subsequently immunized with these glioma extracts. B-cell hybridomas from these mice were then cloned and subcloned by limiting dilution, yielding six monoclonal antibodies (MAbs) that were entirely specific for tumor tissues, and did not react with normal human serum or with normal human brain, liver, kidney, spleen, or muscle. Moreover, the murine MAbs did not cross-react with certain other human tumors, including melanoma. The fully absorbed antiserum and the murine MAbs both identify a polypeptide pattern possibly related to human glial fibrillary acidic protein (GFAP) or other intermediate filament proteins on immunoblots. These immunological reagents could serve as powerful tools for the diagnosis and possibly therapy of these uniformly fatal tumors.

Keywords

Human Serum Albumin Glial Fibrillary Acidic Protein Glioblastoma Multiforme Anaplastic Astrocytoma Normal Brain Tissue 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Wikstrand, C.J. and Bigner, D.D., Surface antigens of human glioma cells shared with normal adult and fetal brain, Cancer Res. 39:3235 (1979).PubMedGoogle Scholar
  2. 2.
    Birkmayer, G.D. and Stass, H.P., Humoral immune response in glioma patients: a solubilized glioma-associated membrane antigen as a tool for detecting circulating antibodies, Int. J. Cancer 25:445 (1980).PubMedCrossRefGoogle Scholar
  3. 3.
    Schnegg, J.F., de Tribolet, N., Diserens, A.-C, Martin-Ashard, A., and Carrel, S., Characterization of a rabbit anti-human malignant glioma antiserum, Int. J. Cancer 28:265 (1981).PubMedCrossRefGoogle Scholar
  4. 4.
    Lowry, O.H., Rosenbrough, N.J., Farr, A.L., and Randall, R.J., Protein measurement with the Folin phenol reagent, J. Biol Chem. 193:265 (1951).PubMedGoogle Scholar
  5. 5.
    Atassi, M.Z., Preparation of monoclonal antibodies to preselected protein regions, Methods in Enzymol. 121:69 (1986).CrossRefGoogle Scholar
  6. 6.
    Hunter, W.M., 1969, V. 608, In: Handbook of Experimental Immunology, D.M. Weir, ed., Davis, Philadelphia.Google Scholar
  7. 7.
    Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature 227:680 (1970).PubMedCrossRefGoogle Scholar
  8. 8.
    Merril, C.R., Goldman, D., Van Keuren, M.L., Simplified silver protein detection and image enhancement in Polyacrylamide gels, Electrophoresis 3:17 (1982).CrossRefGoogle Scholar
  9. 9.
    Towbin, H., Staehelin, T. and Gordon, J., Electrophoretic transfer of proteins from Polyacrylamide gels to nitrocellulose sheets: procedure and some applications, Proc. Natl Acad. Sci. USA 76:4350 (1979).PubMedCrossRefGoogle Scholar
  10. 10.
    Burnette, W.N., Western blotting: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated Protein A, Anal. Biochem. 112:195 (1981).PubMedCrossRefGoogle Scholar
  11. 11.
    Twining, S.S. and Atassi, M.Z., Use of immunoadsorbents for the study of antibody binding to sperm-whale myoglobin and its synthetic antigenic sites, J. Immunol. Methods 30:139 (1979).PubMedCrossRefGoogle Scholar
  12. 12.
    Sakata, S., and Atassi, M.Z., Immune recognition of serum albumin. 13. Auto-reactivity with rabbit serum albumin of rabbit antibodies against bovine or human serum albumins and autoimmune recognition of rabbit serum albumin, Mol Immunol 18:961 (1981).PubMedCrossRefGoogle Scholar
  13. 13.
    Schmitz, H.E., Atassi, H. and Atassi, M.Z., Production of monoclonal antibodies with preselected submolecular specificities: demonstration with sperm-whale myoglobin, Mol Immunol 19:1699 (1982).PubMedCrossRefGoogle Scholar
  14. 14.
    Fischer, D.K., Miller, G., Gradoville, L., Heston, L., Westrate, M.W., Wright, J., Brandsma, J. and Summers, W.C., Genome of a mononucleosis Epstein-Barr virus contains DNA fragments previously regarded to be unique to Burkitt’s lymphoma isolates, Cell 24:543 (1981).PubMedCrossRefGoogle Scholar
  15. 15.
    Silverberg, E. and Lubera, J.A., Cancer statistics, Ca-A Cancer J. for Clinicians 38:5 (1988).CrossRefGoogle Scholar
  16. 16.
    Walker, M.D., Green, S.B., Byar, D.P., Alexander, E., Batzdorf, U., Brooks, W.H., Hunt, W.E., MacCarty, C.S., Mahaley, M.S., Mealey, J., Owens, G., Ransohoff, J., Robertson, J.T., Shapiro, W.R., Smith, K.R., Wilson, C.B. and Strike, T.A., Randomized comparisons of radiotherapy and nitrosoureas for the treatment of malignant glioma after surgery, N Engl J. tMed. 303:1323 (1980).CrossRefGoogle Scholar
  17. 17.
    Narayan, R.K., Heydorn, W.E., Creed, G.J. and Jacobowitz, D.M., Protein patterns in various malignant human brain tumors by two-dimensional gel electrophoresis, Cancer Res. 46:4685 (1986).PubMedGoogle Scholar
  18. 18.
    Schnegg, J.F., Diserens, A.C., Carrel., S., Accolla, R.S. and de Tribolet, N., Human glioma-associated antigens detected by monoclonal antibodies, Cancer Res. 41:1209 (1981).PubMedGoogle Scholar
  19. 19.
    Cairncross, J.G., Mattes, M.J., Beresford, H.R., Albino, A.P., Houghton, A.N., Lloyd, K.O. and Old, L.J., Cell surface antigens of human astrocytoma defined by mouse monoclonal antibodies: identification of astrocytoma subsets, Proc. Natl. Acad. Sci. USA 79:5641 (1982).PubMedCrossRefGoogle Scholar
  20. 20.
    Carrel, S., de Tribolet, N. and Mach, J.P., Expression of neuroectodermal antigens common to melanomas, gliomas, and neuroblastomas. I. Identification by monoclonal anti-melanoma and anti-glioma antibodies, Acta Neuropathol. 57:158 (1982).PubMedCrossRefGoogle Scholar
  21. 21.
    Bourdon, M.A, Wikstrand, C.J., Furthmayr, H., Matthews, T.J. and Bigner, D.B., Human glioma-mesenchymal extracellular matrix antigen defined by monoclonal antibody, Cancer Res. 43:2796 (1983).PubMedGoogle Scholar
  22. 22.
    Jones, D., Fritschy, J., Garson, J., Nokes, T.J.C., Kemshead, J.T. and Hardistry, R.M., A monoclonal antibody binding to human medulloblastoma cells and to the platelet glycoprotein IIb-IIIa complex, Br. J. Haematol. 57:621 (1984).PubMedGoogle Scholar
  23. 23.
    Fischer, D.K., Chen, T.L., and Narayan, R.K., Immunological and biochemical strategies for the identification of brain tumor-associated antigens, J. Neurosurg. 68:165 (1988).PubMedCrossRefGoogle Scholar
  24. 24.
    Coakham, H, Surface antigen(s) common to human astrocytoma cells, Nature 250:328 (1974).PubMedCrossRefGoogle Scholar
  25. 25.
    Wahlstrom, T., Linder, E., Saksela, E. and Westermark, B., Tumor-specific membrane antigens in established cell lines from gliomas, Cancer 34:274 (1974).PubMedCrossRefGoogle Scholar
  26. 26.
    Coakham, H.B. and Lakshmi, M.S., Tumor-associated surface antigen(s) in human astrocytomas, Oncology 31:233 (1975).CrossRefGoogle Scholar
  27. 27.
    Narayan, R.K., Heydorn, W.E., Creed, G.J., Kornblith, P.L. and Jacobowitz, D.M., 2. Two-dimensional gel electrophoretic protein patterns in high-grade human astrocytomas. In: Biology of Brain Tumours, Walker, M.D. and Thomas, D.G., eds., Martinus Nijhoff Publishers, Boston (1986).Google Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Masafumi Matsuda
    • 2
  • Duncan K. Fischer
    • 2
  • Raj K. Narayan
    • 2
  • M. Zouhair Atassi
    • 1
  1. 1.Department of BiochemistryBaylor College of MedicineHoustonUSA
  2. 2.Department of NeurosurgeryBaylor College of MedicineHoustonUSA

Personalised recommendations