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. 1989 Jul;29(3):179–184. doi: 10.1007/BF00199993

Antibody response after immunization with the gangliosides GM1, GM2, GM3, GD2 and GD3 in the mouse

Philip O Livingston 1,✉, Gerd Ritter 1, Michele Jones Calves 1
PMCID: PMC11038927  PMID: 2731184

Abstract

The gangliosides GM2, GD2 and GD3 are differentiation antigens expressed on the cell surface of human melanomas and other cancers of neuroectodermal origin. We have compared the antibody response after vaccination with gangliosides GM1, GM2, GM3, GD2 and GD3 in the mouse. Purified gangliosides were injected subcutaneously alone or attached to Salmonella minnesota mutant R595 after pretreatment of the mice with low-dose cyclophosphamide. Spontaneous GM1 antibodies, but not antibodies against the other gangliosides, were detected in many mice, the incidence increasing with age. Purified gangliosides injected alone (in saline) induced no antibody response. R595/GM1 and R595/GD3 vaccination induced consistent high-titer antibody responses. R595/GM2 and R595/GD2 induced occasional antibody responses, and R595/GM3 induced no antibody response. Comparison of the antibody responses induced against these gangliosides in the mouse with those in man reveals that GM1, GM3 and GD2 have a similar immunogenicity in both species while the relative immunogenicity of GM2 and GD3 is reversed. To understand better the basis for these differences, the antibody responses against the five gangliosides in man and the mouse were compared with their known expression in normal tissues. No correlation was detected between ganglioside expression in normal brain and immunogenicity, consistent with this being a cloistered site. The antibody responses did correlate inversely with expression in normal non-brain human and murine tissues. Variations between species of ganglioside immunogenicity may reflect variations in ganglioside expression in normal tissues.

Keywords: Melanoma, Cell Surface, Cancer Research, Normal Tissue, Cyclophosphamide

Footnotes

Abbreviations used. The abbreviations for gangliosides are according to the system of Svennerholm [43]; ELISA, enzyme-linked immunosorbent assay; R595, S. minnesota rought mutant R595

References

  • 1.Ando S, Chang N-C, Yu RK. High-performance thinlayer chromatography and densitometric determination of brain ganglioside compositions of several species. Anal Biochem. 1978;89:437–450. doi: 10.1016/0003-2697(78)90373-1. [DOI] [PubMed] [Google Scholar]
  • 2.Brightman MW, Reese TS. Junctions between intimately apposed cell membranes in the vertebrate brain. J Cell Biol. 1969;40:648–677. doi: 10.1083/jcb.40.3.648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Cahan LD, Irie RF, Singh R, Cassidenti A, Paulson JC. Identification of a human neuroectodermal tumor antigen (OFA-I-2) as ganglioside GD2. Proc Natl Acad Sci USA. 1982;79:7629. doi: 10.1073/pnas.79.24.7629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Cheresh DA, Harper JR, Schulz G, Reisfeld RA. Localization of the gangliosides GD2 and GD3 in adhesion plaques and on the surface of human melanoma cells. Proc Natl Acad Sci USA. 1984;81:5767. doi: 10.1073/pnas.81.18.5767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Cheresh DA, Honsik CJ, Staffileno LK, Jung G, Reisfeld RA. Disialoganglioside GD3 on human melanoma serves as a relevant target antigen for monoclonal antibody-mediated tumor cytolysis. Proc Natl Acad Sci USA. 1985;82:5155. doi: 10.1073/pnas.82.15.5155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Cheung N-K V, Saarinen UM, Neely JE, Landmeier B, Donovan D, Coccia PF. Monoclonal antibodies to a glycolipid antigen on human neuroblastoma cells. Cancer Res. 1985;45:2642. [PubMed] [Google Scholar]
  • 7.Cheung N-K V, Lazarus H, Miraldi FD, et al. (1987) Ganglioside GD2 specific monoclonal antibody 3F8: a phase I study in patients with neuroblastoma and malignant melanoma. J Clin Oncol 1430–1440 [DOI] [PubMed]
  • 8.Cordon-Cardo C, Graus F, Andersen N, Houghton A, Cheung NK (1988) Distribution of GD2 ganglioside in the human nervous system detected by a mouse monoclonal antibody: relevance to treatment with anti-ganglioside antibodies. Brain Res (in press) [DOI] [PubMed]
  • 9.Dippold WG, Lloyd KO, Li LTC, Ikeda H, Oettgen HF, Old LJ. Cell surface antigens of human malignant melanoma. Proc Natl Acad Sci USA. 1980;77:6114. doi: 10.1073/pnas.77.10.6114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Ehrlich P. On autoimmunity with special references to cell life. Proc R Soc Lond. 1900;66B:424–448. [Google Scholar]
  • 11.Endo T, Scott DD, Stewart SS, Kundu SK, Marcus DM. Antibodies to glycosphinogolipids in patients with multiple sclerosis and SLE. J Immunol. 1980;132:1793–1797. [PubMed] [Google Scholar]
  • 12.Hamanaka S, Handa S, Yamakawa T. Ganglioside compositions of erythrocytes from various strains of inbred mice. J Biochem. 1979;86:1623. doi: 10.1093/oxfordjournals.jbchem.a132681. [DOI] [PubMed] [Google Scholar]
  • 13.Hashimoto Y, Otsuka H, Yamakawa T. The occurrence of GM4 and GM2 in erythrocytes from inbred strains of mice. J Biochem. 1982;91:1039–1046. doi: 10.1093/oxfordjournals.jbchem.a133753. [DOI] [PubMed] [Google Scholar]
  • 14.Hashimoto Y, Suzuki A, Yamakawa T, Miyashita N, Moriwaki K. Expression of GM1 and GD1a in mouse liver is linked to the H-2 complex on chromosome 17. J Biochem. 1983;94:2043–2048. doi: 10.1093/oxfordjournals.jbchem.a134559. [DOI] [PubMed] [Google Scholar]
  • 15.Hawkes R, Niday E, Gordon J. A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem. 1982;119:142–147. doi: 10.1016/0003-2697(82)90677-7. [DOI] [PubMed] [Google Scholar]
  • 16.Head JR, Billingham RE. Immunologically privileged sites in transplantation immunology and oncology. Perspect Biol Med. 1985;29:115–131. doi: 10.1353/pbm.1985.0038. [DOI] [PubMed] [Google Scholar]
  • 17.Herlyn M, Rodeck U, Mancianti M-L, et al. Expression of melanoma-associated antigens in rapidly dividing human melanocytes in culture. Cancer Res. 1987;47:3057–3061. [PubMed] [Google Scholar]
  • 18.Houghton AN, Mintzer D, Cordon-Cardo C, et al. Mouse monoclonal IgG3 antibody detecting GD3 ganglioside: a phase I trial in patients with malignant melanoma. Proc Natl Acad Sci USA. 1985;82:1242. doi: 10.1073/pnas.82.4.1242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Iwamori M, Harpin ML, Lachapelle F, Baumann N. Brain gangliosides of quaking and shiverer mutants: qualitative and quantitative changes of monosialogangliosides in the quaking brain. J Neurochem. 1985;45:73–78. doi: 10.1111/j.1471-4159.1985.tb05476.x. [DOI] [PubMed] [Google Scholar]
  • 20.Kawashima I, Tada N, Ikegami S, Nakamura S, Ueda R, Tai T. Mouse monoclonal antibodies detecting disialogangliosides on mouse and human T lymphomas. Int J Cancer. 1988;41:267–274. doi: 10.1002/ijc.2910410218. [DOI] [PubMed] [Google Scholar]
  • 21.Kemp JD, Weinfeld HF, Koerner TAW. A quantitative analysis of H-2 linked effects on hepatic ganglioside composition. Immunogenetics. 1987;26:130–137. doi: 10.1007/BF00365901. [DOI] [PubMed] [Google Scholar]
  • 22.Livingston PO. Experimental and clinical studies with active specific immunotherapy. In: Mitchell MS, editor. Immunity to cancer II. New York: Liss; 1989. pp. 309–322. [PubMed] [Google Scholar]
  • 23.Livingston PO, DeLeo AB, Jones M, Oettgen HF, Old LJ. Comparison of approaches for augmenting the serological response to the Meth A antigen. Pretreatment with cyclophosphamide is most effective. J Immunol. 1983;131:2601. [PubMed] [Google Scholar]
  • 24.Livingston PO, Jones M, DeLeo AB, Oettgen HF, Old LJ. The serological response to Meth A sarcoma vaccines after cyclophosphamide treatment is further increased by various adjuvants. J Immunol. 1985;135:1505–1509. [PubMed] [Google Scholar]
  • 25.Livingston PO, Kaelin E, Pinsky CM, Oettgen HF, Old LJ. IV. Serological response in stage II melanoma patients receiving allogeneic melanoma cell vaccines. Cancer. 1985;56:2194–2220. doi: 10.1002/1097-0142(19851101)56:9<2194::aid-cncr2820560910>3.0.co;2-l. [DOI] [PubMed] [Google Scholar]
  • 26.Livingston PO, Jones Calves M, Natoli EJ., Jr Approaches to augmenting the immunogenicity of the ganglioside GM2 in mice: Purified GM2 is superior to whole cells. J Immunol. 1987;138:1524–1529. [PubMed] [Google Scholar]
  • 27.Livingston PO, Natoli EJ, Jr, Jones Calves M, Stockert E, Oettgen HF, Old LJ. Vaccines containing purified GM2 ganglioside elicit GM2 antibodies in melanoma patients. Proc Natl Acad Sci USA. 1987;84:2911–2915. doi: 10.1073/pnas.84.9.2911. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Mahadik S, Laev H, Rapport MM. Preparation and specificity of 11 monoclonal antibodies to GM1 ganglioside. J Neurochem. 1986;47:1172–1175. doi: 10.1111/j.1471-4159.1986.tb00736.x. [DOI] [PubMed] [Google Scholar]
  • 29.Milgrom F. Allotransplantation and autoimmunity. Transplant Proc. 1980;12:557–565. [PubMed] [Google Scholar]
  • 30.Miyake M, Ito M, Hitomi S, et al. Generation of two murine monoclonal antibodies that can discriminate N-glycolyl and N-acetyl neuraminic acid residues of GM2 gangliosides. Cancer Res. 1988;48:6154–6160. [PubMed] [Google Scholar]
  • 31.Nakabayashi H, Iwamori M, Nagai Y. Analysis and quantitation of gangliosides as p-bromophenacyl derivatives by high-performance liquid chromatography. J Biochem. 1984;96:977–984. [PubMed] [Google Scholar]
  • 32.Nakamura K, Hashimoto Y, Yamakawa T, Suzuki A. Genetic polymorphism of ganglioside expression in mouse organs. J Biochem. 1988;103:201–208. doi: 10.1093/oxfordjournals.jbchem.a122232. [DOI] [PubMed] [Google Scholar]
  • 33.Natoli EJ, Jr, Livingston PO, Cordon-Cardo C, et al. A murine monoclonal antibody detecting N-acetyl and N-glycolyl GM2: characterization of cell surface reactivity. Cancer Res. 1986;46:4116–4120. [PubMed] [Google Scholar]
  • 34.Nilsson O, Svennerholm L. Characterization and quantitative determination of gangliosides and neutral glycosphingolipids in human liver. J Lipid Res. 1982;23:327–334. [PubMed] [Google Scholar]
  • 35.Nores GA, Dohi T, Taniguchi M, Hakomori S-I. Density-dependent recognition of cell surface GM3 by a certain anti-melanoma antibody, and GM3 lactone as a possible immunogen: requirements for tumor-associated antigen and immunogen. J Immunol. 1987;139:3171–3176. [PubMed] [Google Scholar]
  • 36.Nudelman ES, Hakomori S, Kannagi R, Levery S, Yeh M-H, Hellstrom KE, Hellstrom I. Characterization of a human melanoma-associated antigen defined by a monoclonal antibody, 4.2. J Biol chem. 1982;257:12752. [PubMed] [Google Scholar]
  • 37.Portoukalian J, Zwingelstein N A-M, Dore J-F. Alteration of gangliosides in plasma and red cells of humans bearing melanoma tumors. Biochem Biophys Commun. 1978;85:916–920. doi: 10.1016/0006-291x(78)90630-7. [DOI] [PubMed] [Google Scholar]
  • 38.Rauvala H. Isolation and partial characterization of human kidney gangliosides. Biochim Biophys Acta. 1976;424:284–295. doi: 10.1016/0005-2760(76)90196-x. [DOI] [PubMed] [Google Scholar]
  • 39.Real FX, Houghton AN, Albino AP, Cordon-Cardo C, Melamed MR, Oettgen HF, Old LJ. Surface antigens of melanomas and melanocytes defined by mouse monoclonal antibodies: specificity analysis and comparison of antigen expression in cultured cells and tissues. Cancer Res. 1985;45:4401–4411. [PubMed] [Google Scholar]
  • 40.Richards RL, Alving CR. Immune reactivities of antibodies against glycolipids. Am Cancer Soc. 1980;27:461–473. [Google Scholar]
  • 41.Rose NR. Autoimmune diseases. Sci Am. 1981;244:80–103. doi: 10.1038/scientificamerican0281-80. [DOI] [PubMed] [Google Scholar]
  • 42.Seyfried TN, Glaser GH, Yu RK. Genetic variability for regional brain gangliosides in five strains of young mice. Biochem Genet. 1979;17:43. doi: 10.1007/BF00484473. [DOI] [PubMed] [Google Scholar]
  • 43.Svennerholm L. Chromatographic separation of human brain gangliosides. J Neurochem. 1963;10:613–623. doi: 10.1111/j.1471-4159.1963.tb08933.x. [DOI] [PubMed] [Google Scholar]
  • 44.Tai T, Cahan LD, Tsuchida T, Saxton RE, Irie RF, Morton DL. Immunogenicity of melanoma-associated gangliosides in cancer patients. Int J Cancer. 1985;35:607. doi: 10.1002/ijc.2910350507. [DOI] [PubMed] [Google Scholar]
  • 45.Taniguchi M, Wakabayashi S. Shared antigenic determinant expressed on various mammalian melanoma cells. Gan. 1984;75:418. [PubMed] [Google Scholar]
  • 46.Towbin H, Schoenenberger C, Ball R, Braun DG, Roselfelder G. Glycosphingolipid-blotting: an immunological detection procedure after separation by thin layer chromatography. J Immunol Methods. 1984;72:471–479. doi: 10.1016/0022-1759(84)90015-2. [DOI] [PubMed] [Google Scholar]
  • 47.Tsuchida T, Saxton RE, Irie RF. Gangliosides of human melanoma: GM2 and tumorigenicity. J Natl Cancer Inst. 1987;78:55–59. doi: 10.1093/jnci/78.1.55. [DOI] [PubMed] [Google Scholar]
  • 48.Welte K, Miller G, Chapman PB, Natoli E, Kunicka JE, Cordon-Cardo C, Old LJ, Houghton AN. Stimulation of T lymphocyte proliferation by monoclonal antibodies against GD3 ganglioside. J Immunol. 1987;139:1763–1771. [PubMed] [Google Scholar]
  • 49.Werkmeister JA, Triglia T, Mackay IR, Dowling JP, Varigos GA, Morstyn G, Burns GF. Fluctuations in the expression of a glycolipid antigen associated with differentiation of melanoma cells monitored by a monoclonal antibody. Leo Mel 31 . Cancer Res. 1987;47:225–230. [PubMed] [Google Scholar]
  • 50.Wiegandt H. Gangliosides of extraneural organs. Hoppe-Seyler's Z Physiol Chem. 1973;354:1049–1056. doi: 10.1515/bchm2.1973.354.2.1049. [DOI] [PubMed] [Google Scholar]

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