Summary
The hybridoma technique was used to generate monoclonal antibodies against a wide spectrum of melanoma-associated surface antigens. Mice were immunized against the human melanoma lines Mel A-375, SK Mel-25, and Mel S-5 (subclone of SK Mel-25), which differ with respect to a number of biological and biochemical properties. Spleen cells were fused with P3×63-AG8.653 myeloma cells. Twenty hybridomas producing antibodies that were negative on platelets, leukocytes, and monocytes but positive on melanoma cells were isolated and recloned. The specificity of antibodies was investigated on 30 human melanoma and nonmelanoma lines. Five groups of antibodies could be distinguished by their reactivity (1) with few melanoma lines and embryonic fibroblasts; (2) with melanoma, neuroblastoma, and teratoma; (3) with melanoma, neuroblastoma, glioblastoma, teratoma, and carcinoma; (4) with melanoma, teratoma, and carcinoma; and (5) with melanoma, neuroblastoma, teratoma, glioblastoma, carcinoma, embryonic fibroblasts, and B-lymphoblastoid cells. The antigen expression was qualitatively and quantitatively different from cell line to cell line. No evidence for melanoma-specific antigens was found. Eight antibodies were isolated detecting phenotypic differences on sublines of SK Mel-25.
Similar content being viewed by others
References
Brüggen J, Sorg C, Macher E (1978) Membrane-associated antigens of human malignant melanoma. V. Serological typing of cell lines using antisera from nonhuman primates. Cancer Immunol Immunother 5:53
Brüggen J, Macher E, Sorg C (1981a) Expression of surface antigens and its relation to parameters of malignancy in human malignant melanoma. Cancer Immunol Immunother 10:121
Brüggen J, Fogh J, Sorg C (1981b) Tumor production in the nude mouse fibrinolytic activity and cross-reactivity with anti-melanoma sera of various human tumor cell lines. J Cancer Res Clin Oncol 102:141
Carrel S, Accolla RS, Carmagnola AL, Mach J-P (1980) Common human melanoma-associated antigen(s) detected by monoclonal antibodies. Cancer Res 40:2523
Dippold WG, Lloyd KO, Li LTC, Ikeda H, Oettgen HF, Old LJ (1980) Cell surface antigens of human malignant melanoma: Definition of six antigenic systems with mouse monoclonal antibodies. Prog Natl Acad Sci USA 77:6114
Feige U, Sorg C (1982) Collection of large quantitites of mononuclear cells from individual donors by continuous flow centrifugation leukapheresis. Clin Exp Immunol
Fogh J, Fogh H (1969) Procedures for control of mycoplasma contamination of tissue cultures. Ann NY Acad Sci 172:15
Gaynor R, Irie R, Morton D, Herschman HR (1980) S100 protein is present in cultured human malignant melanomas. Nature 286:400
Gefter ML, Margulies DH, Scharff MO (1977) A simple method for polyethyleneglycol promoted hybridization of mouse myeloma cells. Somatic Cell Genet 3:231
Herlyn M, Clark WH Jr, Mastrangelo MJ, Guerry D IV, Elder DE, LaRossa D, Hamilton R, Bondi E, Tuthill R, Steplewski Z, Koprowski H (1980) Specific immunoreactivity of hybridoma-secreted monoclonal anti-melanoma antibodies to cultured cells and freshly derived human cells. Cancer Res 40:3602
Imai K, Ng A-K, Ferrone S (1981) Characterization of monoclonal antibodies to human melanoma-associated antigens. J Natl Cancer Inst 66:489
Johnson JP, Demmer-Dieckmann M, Meo T, Hadam MR, Riethmüller G (1981) Surface antigens of human melanoma cells defined by monoclonal antibodies. I. Biochemical characterization of two antigens found on cell lines and fresh tumors of diverse tissue origin. Eur J Immunol 11:825
Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495
Müller C, Sorg C (1975) Use of formalin-fixed melanoma cells for the detection of antibodies against surface antigens by a micro-immune adherence technique. Eur J Immunol 5:175
Natali PG, Wilson BS, Imai K, Bigotti A, Ferrone S (1982) Tissue distribution, molecular profile, and shedding of a cytoplasmic antigen identified by the monoclonal antibody 465.12S to human melanoma cells. Cancer Res 42:583
Nicolson GL, Birdwell CR, Brunson KW, Robbins JC, Beattie L, Fidler IJ (1977) Cell interactions in the metastatic process: some cell surface properties associated with successful bloodborne tumor spread. In: Lash JW, Burger MM (eds) Cell and tissue interactions. Raven Press, New York, p 225
Pierce GB, Wallace C (1971) Differentiation of malignant to benign cells. Cancer Res 31:127
Poste G, Fidler JJ (1980) The pathogenesis of cancer metastasis. Nature 283:139
Seeger RC, Rosenblatt HM, Imai K, Ferrone S (1981) Common antigenic determinants on human melanoma, glioma, neuroblastoma, and sarcoma cells defined with monoclonal antibodies. Cancer Res 41:2714
Shiku H, Takahashi OF, Oettgen HF, Old LJ (1976) Cell surface antigens of human malignant melanoma. II. Serological typing with immune adherence assays and definition of two new surface antigens. J Exp Med 144:873
Sorg C, Brüggen J, Seibert E, Macher E (1978) Membrane-associated antigens of human malignant melanoma. IV. Changes in expression of antigens on cultured melanoma cells. Cancer Immunol Immunother 3:259
Stenzinger W, Brüggen J, Macher E, Sorg C (1983) Tumor angiogenic activity (TAA) production in vitro and growth in the nude mouse by human malignant melanoma. Eur J Cancer Clin Oncol 19:649
Stuhlmiller GM, Seigler HF (1975) Characterization of a chimpanzee antihuman melanoma antiserum. Cancer Res 35:2132
Suter L, Brüggen J, Sorg C (1980) Use of an enzyme-linked immunosorbent assay (ELISA) for screening of hybridoma antibodies against cell surface antigens. J Immunol Meth 39:407
Wilkstrand 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
Wilson BS, Indiveri F, Bellegrino MA, Ferrone S (1979) DR (Ia-like) antigens on human melanoma cells: serological detection and immunological characterization. J Exp Med 149:658
Winchester RJ, Wang CY, Gebofsky A, Vunkel HG, Lloyd KO, Old LJ (1978) Expression of the Ia-like antigens on cultured human malignant melanoma cell lines. Proc Natl Acad Sci USA 75:6235
Woodbury RG, Brown JP, Yeh, M-Y, Hellström I, Hellström KE (1980) Identification of a cell surface protein, p 97, in human melanomas and certain other neoplasms. Proc Natl Acad Sci USA 77:2183
Yeh M-Y, Hellström I, Brown JP, Warner GA, Hansen JA, Hellström KE (1979) Cell surface antigens of human melanoma identified by monoclonal antibody. Proc Natl Acad Sci USA 76:2927
Yeh M-Y, Hellström I, Hellström KE (1981) Clonal variation in expression of a human melanoma antigen defined by a monoclonal antibody. J Immunol 126:1312
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Brüggen, J., Sorg, C. Detection of phenotypic differences on human malignant melanoma lines and their variant sublines with monoclonal antibodies. Cancer Immunol Immunother 15, 200–205 (1983). https://doi.org/10.1007/BF00199165
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00199165