Summary
Monoclonal antibodies (mAbs) recognizing the disialoganglioside II3(NeuAc)2GgOse3Cer (GD2) were produced by immunizing mice with the GD2-expressing neuroblastoma cell line LAN-1 and a prefusion boost with purified GD2 coupled to Salmonella minnesota. Two IgM mAbs were isolated which demonstrated high levels of reactivity (binding ratios in excess of 100) with GD2 by solid-phase radioimmunoassay and positivity in high-performance thin-layer chromatography (HPTLC) immunostain; only one (DMAb-20) was subsequently shown by analysis with a panel of defined ganglioside species to be specific for the minimum epitope of GD2, GalNAcβ1-4(NeuAcα2-8NeuAcα2-3)Gal-. DMAb-20 was used to evaluate the expression of GD2 by malignant glioma and medulloblastoma cell lines using cell surface radioimmunoassay, indirect membrane immunofluorescence, HPTLC immunostain, and densitometric analysis of extracted gangliosides from selected cell lines. Sixteen of 20 (80%) malignant glioma and 5 of 5 medulloblastoma cell lines reacted with DMAb-20; in agreement with previous studies, 5 of 5 neuroblastoma and 2 of 3 melanoma cell lines also reacted with DMAb-20. GD2 was proportionally increased in the glioma and medulloblastoma cell lines relative to levels in normal brain, as determined by densitometric analysis. In a phenotypic survey of malignant glioma biopsies, tumor cells in 24 of 30 (80%) cases stained positively with DMAb-20. Reactive astrocytes, both within and adjacent to tumors, were frequently intensely stained. Among the morphological variants of glioblastoma examined, the most intense staining with DMAb-20 was observed in neoplastic gemistocytes, with the weakest or absent staining in small cell glioblastomas. As GD2 is a commonly expressed surface antigen of gliomas and medulloblastomas, expression of which is retained in tissue culture, DMAb-20 will be useful in determining the functional role of GD2 in cell-cell interaction, adhesion, and invasion, and in defining altered growth control mechanisms of central nervous system neoplasms in in vitro models.
Similar content being viewed by others
Abbreviations
- xxGangliosides have been designated according to CBN recommendations [22] and to the coding system of Svennerholm [35] GD2:
-
II3(NeuAc)2GgOse3Cer
- GM3:
-
II3NeuAc-LacCer
- GD3:
-
II3(NeuAc)2-LacCer
- GM2:
-
II3NeuAcGgOse3Cer
- GM1:
-
II3NeuAcGgOse4Cer
- GD1a:
-
II3NeuAcIV3NeuAcGgOse4Cer
- GD1b:
-
II3(NeuAc)2GgOse4Cer
- GT1b:
-
IV3NeuAcII3(NeuAc)2GgOse4Cer
- GQ1b:
-
IV3(NeuAc)2II3(NeuAc)2GgOse4Cer
- 3′,8′-LD1:
-
IV3(NeuAc)2nLeOse4Cer
References
Ando S, Chang N-C, Yu RK (1978) High-performance thinlayer chromatography and densitometric determination of brain ganglioside composition of several species. Anal Biochem 89:437–450
Bigner DD, Bigner SH, Pontén J, Westermark B, Mahaley MS Jr, Ruoslahti E, Herschman H, Eng LF, Wikstrand CJ (1981) Heterogeneity of genotypic and phenotypic characteristics of fifteen permanent cell lines derived from human gliomas. J Neuropathol Exp Neurol 40:201–229
Bigner SH, Friedman HS, Vogelstein B, Oakes WJ, Bigner DD (1990) Amplification of the c-myc gene in medulloblastoma cell lines and xenografts. Cancer Res 50:2347–2350
Bosslet K, Mennel HD, Rodden F, Bauer BL, Wagner F, Altmannsberger A, Sedlacek HH, Wiegandt H (1989) Monoclonal antibodies against epitopes on ganglioside GD2 and its lactones: markers for gliomas and neuroblastomas. Cancer Immunol Immunother 29:171–178
Bourdon MA, Wikstrand CJ, Furthmayr H, Matthews TJ, Bigner DD (1983) Human glioma-mesenchymal extracellular matrix antigen defined by monoclonal antibody. Cancer Res 43:2796–2805
Cahan LD, Irie RF, Singh R, Cassidenti A, Paulson JC (1982) Identification of a human neuroectodermal tumor antigen (OFA-I-2) as ganglioside GD2. Proc Natl Acad Sci USA 79:7629–7633
Cheresh DA, Harper JR, Schulz G, Reisfeld RA (1984) Localization of the gangliosides GD2 and GD3 in adhesion plaques and on the surface of human melanoma cells. Proc Natl Acad Sci USA 81:5767–5771
Cheung N-KV, Saarinen UM, Neely JE, Landmeier B, Donovan D, Coccia PF (1985) Monoclonal antibodies to a glycolipid antigen on human neuroblastoma cells. Cancer Res 45:2642–2649
Cohen ME, Duffner PK (1984) Brain tumors in children. Principles of diagnosis and treatment. Raven Press, New York, pp 1–8
Dawson G, Stefansson K (1984) Gangliosides of human spinal cord: aberrant composition of cords from patients with amyotrophic lateral sclerosis. J Neurosci Res 12:213–220
Distler JJ, Jourdian GW (1973) The purification and properties of β-galactosidase from bovine testes. J Biol Chem 248:6672–6680
Fredman P, Magnani JL, Nirenberg M, Ginsburg V (1984) Monoclonal antibody A2B5 reacts with many gangliosides in neuronal tissue. Arch Biochem Biophys 233:661–666
Fredman P, von Holst H, Collins VP, Ammar A, Dellheden B, Wahren B, Granholm L, Svennerholm L (1986) Potential ganglioside antigens associated with human gliomas. Neurol Res 8:123–126
Fredman P, M»nsson J-E, Bigner S, Wikstrand C, Bigner D, Svennerholm L (1990) Gangliosides in the human ghoma cell line U-118 MG grown in culture or as xenografts in nude rats. Biochim Biophys Acta 1045:239–244
Friedman HS, Burger PC, Bigner SH, Trojanowski JW, Wikstrand CJ, Halperin EC Bigner DD (1985) Establishment and characterization of the human medulloblastoma cell line and transplantable xenograft D283 Med. J Neuropathol Exp Neurol 44:592–605
Friedman HS, Burger PC, Bigner SH, Trojanowski JW, Brodeur GM, He X, Wikstrand CJ, Kurtzberg J, Berens ME, Halperin EC, Bigner DD (1988) Phenotypic and genotypic analysis of a human medulloblastoma cell line and transplantable xenograft (D341 Med) demonstrating amplification of c-myc. Am J Pathol 130:472–484
Gillard BK, Thomas JW, Nell LJ, Marcus DM (1989) Antibodies against ganglioside GT3 in the sera of patients with type 1 diabetes mellitus. J Immunol 142:3826–3832
Gross N, Beck D, Portoukalian J, Favre S, Carrel S (1989) New anti-GD2 monoclonal antibodies produced from gammainterferon treated neuroblastoma cells. Int J Cancer 43:665–671
Hakomori S (1985) Aberrant glycosylation in cancer cell membranes as focused on glycolipids: overview and perspectives. Cancer Res 45:2405–2414
Heiner JP, Miraldi F, Kallick S, Makley J, Neely J, Smith-Mensah WH, Cheung N-KV (1987) Localization of GD2-specific monoclonal antibody 3F8 in human osteosarcoma. Cancer Res 47:5377–5381
Hsu S-M, Raine L, Fanger H (1981) Use of avidin-biotinperoxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabelled antibody (PAP) procedures. J Histochem Cytochem 29:577–580
IUPAC-IUB Commission on Biochemical Nomenclature (CBN) (1977) The nomenclature of lipids. Eur J Biochem 79:11–21
Jacobsen PF, Jenkyn DJ, Papadimitriou JM (1985) Establishment of a human medulloblastoma cell line and its heterotransplantation into nude mice. J Neuropathol Exp Neurol 44:472–485
Kawashima I, Tada N, Ikegami S, Nakamura S, Ueda R, Tai T (1988) Mouse monoclonal antibodies detecting disialogangliosides on mouse and human T lymphomas. Int J Cancer 41:267–274
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Mujoo K, Cheresh DA, Yang HM, Reisfeld RA (1987) Disialoganglioside GD2 on human neuroblastoma cells: target antigen for monoclonal antibody-mediated cytolysis and suppression of tumor growth. Cancer Res 47:1098–1104
Mujoo K, Kipps TJ, Yang HM, Cheresh DA, Wargalla U, Sander DJ, Reisfeld RA (1989) Functional properties and effect on growth suppression of human neuroblastoma tumors by isotype switch of monoclonal anti-ganglioside GD2 antibody 14.18. Cancer Res 49:2857–2861
Olsson Y, Sourander P, Svennerholm L (1966) Experimental studies on the pathogenesis of leucodystrophies. I. The effect of intracerebrally injected sphingolipids in the rat's brain. Acta Neuropathol (Berl) 6:153–163
Richter A, Sanford KK, Evans VJ (1972) Influence of oxygen and culture media on plating efficiency of some mammalian tissue cells. J Natl Cancer Inst 49:1705–1712
Rosengren B, M»nsson J-E, Svennerholm L (1987) Composition of gangliosides and neutral glycosphingolipids of brain in classical Tay-Sachs and Sandhoff disease: more lyso-GM2 in Sandhoff disease?. J Neurochem 49:834–840
Schulz G, Cheresh DA, Varki NM, Yu A, Staffileno LK, Reisfeld GA (1984) Detection of ganglioside GD2 in tumor tissues and sera of neuroblastoma patients. Cancer Res 44:5914–5920
Slagel DE, Dittmer JC, Wilson CB (1967) Lipid composition of human glial tumor and adjacent brain. J Neurochem 14:789–798
Suzuki K (1965) The pattern of mammalian brain gangliosides. III. Regional and developmental differences. J Neurochem 12:969–979
Svennerholm L (1957) Quantitative estimation of sialic acids. II. A colorimetric resorcinol-hydrochloric acid method. Biochim Biophys Acta 24:604–611
Svennerholm L (1963) Chromatographic separation of human brain gangliosides. J Neurochem 10:613–623
Svennerholm L (1970) Gangliosides. In: Lajtha A (ed) Handbook of neurochemistry, vol 3, Plenum Press, New York, pp 425–452
Svennerholm L, Fredman P (1980) A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta 617:97–109
Tai T, Kawashima I, Tada N, Dairiki K (1988) Different fine binding specificities of monoclonal antibodies to disialoganglioside GD2. J Biochem 103:682–687
Thurin J, Thurin M, Kimoto Y, Herlyn M, Lubeck MD, Elder DE Smereczynski M, Karlson K-A, Clark WM, Steplewski Z, Koprowski H (1987) Monoclonal antibody-defined correlations in melanoma between levels of GD2 and GD3 antigens and antibody-mediated cytotoxicity. Cancer Res 47:1229–1233
Traylor TD, Hogan EL (1980) Gangliosides of human cerebral astrocytomas. J Neurochem 34:126–131
Vrionis FD, Wikstrand CJ, Fredman P, M»nsson J-E, Svennerholm L, Bigner DD (1989) Five new epitope-defined monoclonal antibodies reactive with GM2 and human glioma and medulloblastoma cell lines. Cancer Res 49:6645–6651
Wikstrand CJ, Bigner DD (1982) Expression of human fetal brain antigens by human tumors of neuroectodermal origin as defined by monoclonal antibodies. Cancer Res 42:267–272
Wikstrand CJ, Mahaley MS, Bigner DD (1977) Surface antigenic characteristics of human glial brain tumor cells. Cancer Res 37:4267–4275
Author information
Authors and Affiliations
Additional information
Supported in part by NIH Grants R37 CA 11898, NS 20023, CA 32672, and T32-NS 07304, and by a grant from the Swedish Medical Research Council (Project 03X-627). Dr. Longee is an Association of Medical School Pediatric Department Chairmen, Inc., Pediatric Scientist Training Program Fellow supported by St. Jude Children's Research Hospital
Rights and permissions
About this article
Cite this article
Longee, D.C., Wikstrand, C.J., M»nsson, J.E. et al. Disialoganglioside GD2 in human neuroectodermal tumor cell lines and gliomas. Acta Neuropathol 82, 45–54 (1991). https://doi.org/10.1007/BF00310922
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00310922