Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Binding specificity of two monoclonal antiglioma antibodies

Immunocytochemical studies using a new tissue embedding technique


The binding specificities of two monoclonal antiglioma antibodies (MAB) derived from hybrids GE2 and BF7 (Schnegg et al. 1981) were tested by indirect immunofluorescence (IF) and two immunoperoxidase (IP) methods. Studies were done on biopsies from 33 human CNS tumors, human derived glioma cells, and N-ethylnitrosourea-induced neurogenic rat cells in culture. The immunohistochemical reactions of MAB were investigated in snap-frozen tumor material, conventionally paraffin-embedded material, and tumors embedded in formol sucrose/gum sucrose/paraffin (FSGSP) by the new tissue processing technique of Bolton and Mesnard (1982), which preserves and enhances the antigenicity of tissues. The FSGSP processing offered a better immunocytochemical staining with MAB as compared to cryostat material, while the conventionally embedded paraffin sections of tumors did not stain at all. The binding of MAB revealed an affinity to both glial tumor cells and normal astrocytes. The techniques described are suitable for the identification of an astrocytic subpopulation within gliomas, and may improve the understaining of antigen expression in various stages of astrocytic dedifferentiation.

This is a preview of subscription content, log in to check access.


  1. Bissell MG, Rubinstein LJ, Bignami A, Herman MM (1974) Characteristic of the rat C-6 glioma maintained in organ culture systems. Production of glial fibrillary acidic protein in the absence of gliofibrillogenesis. Brain Res 82:77–89

  2. Bolton WK, Mesnard RB (1982) Methods in laboratory investigation. New technique of kidney tissue processing for immunofluorescence microscopy: formol sucrose/gum sucrose/paraffin. Lab Invest 47:206–213

  3. Chronwall BM, McKeever PE, Kornblith PL (1983) Glial and on-glial neoplasms evaluated on frozen section by double immunofluorescence for fibronectin and glial fibrillary acidic protein. Acta Neuropathol (Berl) 59:283–287

  4. Coakham HB, Kornblith PL, Quindlen EA, Pollack LA, Wood WC, Hartnett LC (1980) Autologous humoral response to human gliomas and analysis of certain cell surface antigens: in vitro study with the use of microcytotoxicity and immune adherence assays. J Natl Cancer Inst 64:223–233

  5. Diserens AC, de Tribolet N, Martin-Achard A, Gaide AC, Schnegg JF, Carrel S (1981) Characterisation of an established human malignant glioma cell line: LN-18. Acta Neuropathol (Berl) 53:21–28

  6. Eggers AE, Tarmin L (1984) In vitro immunisation against autologous glioblastoma cells coupled to adjuvant peptide. J Neurol Sci 63:147–151

  7. Frame MC, Freshney RI, Vaughan PFT, Graham DI, Shaw R (1984) Interrelationship between differentiation and malignancy-associated properties in glioma. Br J Cancer 49:269–280

  8. Gilden DH, Wróblewska Z, Eng LF, Rorke LB (1976) Human brain in tissue culture. Part 5. Identification of glial cells by immunofluorescence. J Neurol Sci 29:177–184

  9. Hitchock MH, Hollinshead AC, Chretien P, Rizzoli HV (1977) Soluble membrane antigens of brain tumors. I. Controlled testing for cell-mediated immune response in a long-surviving glioblastoma multiforme patient. Cancer 40:660–666

  10. Hsu SM, Raine L (1981) Protein A, avidin and biotin in immunohistochemistry. J Histochem Cytochem 29:1349–1353

  11. Jessen KR, Thorpe R, Mirsky R (1984) Molecular identity, distribution and heterogenity of glial fibrillary acidic protein: an immunoblotting and immunohistochemical study of Schwann cells, satellite cells, enteric glia and astrocytes. J Neurocytol 13:187–200

  12. Ko L, Köstner A, Wechsler W (1980) Morphological characterisation of nitrosourea-induced glioma cell lines and clones. Acta Neuropathol (Berl) 51:23–31

  13. Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256: 495–497

  14. Martin-Achard A, Diserens AC, de Tribolet N, Carrel S (1980) Evaluation of the humoral response of glioma patients to a possible common tumor-associated antigen(s). Int J Cancer 25:219–224

  15. Osborn M, Weber K (1983) Biology of disease. Tumor diagnosis by intermediate filament typing: a novel tool for surgical pathology. Lab Invest 48:372–394

  16. Pfeiffer SE, Wechsler W (1972) Biochemically differentiated neoplastic clone of Schwann cells. Proc Natl Acad Sci USA 69:2885–2889

  17. Pfreundschuh M, Shiku H, Takahashi T, Ueda R, Ransohoff J, Oettgen HF, Old LJ (1978) Serological analysis of cell surface antigens of malignant human brain tumors. Proc Natl Acad Sci USA 75:5122–5126

  18. Rosenblum ML, Gerosa MA (1984) Stem cell sensitivity. Prog Exp Tumor Res 28:1–17

  19. Schnegg JF, Diserens AC, Carell S, Accolla RS, de Tribolet N (1981) Human glioma-associated antigens detected by monoclonal antibodies. Cancer Res 41:1209–1213

  20. Stavrou D, Süss C, Bilzer T, Kummer U, de Tribolet N (1983) Monoclonal antibodies reactive with glioma cell lines derived from experimental brain tumors. Eur J Cancer Clin Oncol 19:1439–1449

  21. Stavrou D, Süss C, Bilzer T, Kummer U, de Tribolet N (1984) Glioma-associated antigens detected by monoclonal antibodies. Anticancer Res [Suppl 1] 4:15–18

  22. Sternberger LA, Hardy PH Jr, Cuculis FF, Meyer HG (1970) The unlabelled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antiperoxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333

  23. Tardieu M, Noseworth JH, Perry L, Che M, greene MI, Weiner HL (1983) Generation of a monoclonal antibody (Epenl) which bind selectively to murine ependymal cells. Brain Res 277:339–346

  24. Thompson JJ, Herlyn MF, Elder DE, Clark WH, Steplewski Z, Koprowski H (1982) Use of monoclonal antibodies in detection of melanoma-associated antigens in intact human tumors. Am J Pathol 107:357–361

  25. de Tribolet N, Carrel S, Mach J-P (1984a) Brain tumor-associated antigens. Prog Exp Tumor Res 27:118–131

  26. de Tribolet N, de Muralt B, Buchegger F, Mach J-P, Schreyer M, Carrel S (1984b) Monoclonal antibodies to glioma tumor antigens. In: Wright GL (ed) Monoclonal antibodies and cancer. Dekker, New York Basel, pp 81–120

  27. Wahlström T, Linder E, Saksela E, Westermark B (1974) Tumor-specific membrane antigens in established cell lines from gliomas. Cancer 34:272–279

  28. Weinrauder H, Krajewski S (1979) Localisation of organ-specific brain antigens with homo- and heterologous antisera by immunofluorescence technique. Neuropathol Pol 17:273–285

  29. Weizsäcker M, Rosenblum ML (1982) Chemosensitivity testing of human brain tumors in vitro. In: Brock M (ed) Modern neurosurgery. Springer, Berlin Heidelberg New York, pp 144–151

  30. Westermark B, Ponten J, Hugosson R (1973) Determinants for establishment of permanent tissue culture lines from human gliomas. Acta Pathol Microbiol Scand 81:791

  31. Wikstrand CJ, Bigner DD (1979) Surface antigens of human glioma cells shared with normal adult and fetal brain. Cancer Res 39:3235–3243

Download references

Author information

Correspondence to S. Krajewski or W. Wechsler.

Additional information

Supported by grant no. SFB 200 from the DFG

Dept. of Neuropathology, Medical Research Centre Polish Academy of Sciences, PL-00-784 Warsaw, 3 Dworkowa Str., Poland

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Krajewski, S., Schwendemann, G., Weizsäcker, M. et al. Binding specificity of two monoclonal antiglioma antibodies. Acta Neuropathol 69, 124–131 (1986).

Download citation

Key words

  • Immunocytochemistry
  • Formol sucrose/gum sucrose/paraffin tissue processing
  • Monoclonal antiglioma antibodies