Summary
In studies on antitumor antibody-cytotoxic drug conjugates as potential tumor-selective cytotoxic agents, methotrexate (MTX) was conjugated via its active ester derivative with a murine monoclonal antibody (aMM46) to a mouse mammary tumor antigen (MM antigen) on syngeneic, ascitic C3H/He mouse mammary tumor MM46 cells. The conjugate retained full antibody activity, as assayed by complement-dependent cytolysis. The target-selective cytotoxicity of aMM46-MTX was verified by the observations that this conjugate showed greater cytotoxicity than the corresponding normal mouse immunoglobulin (nIg) conjugate to MM46 cells, neither aMM46 nor nIg being cytotoxic, and that it showed less cytotoxicity to MM antigen negative mouse mammary tumor MM48 cells than to MM46 cells, its cytotoxicity to MM48 cells being similar to that of the nIg conjugate. From the results of assays of cell binding and uptake of 131I-labeled aMM46 and aMM46-3H-MTX, aMM46 and aMM46-MTX were internalized after their binding to MM46 cell surface antigen. Leupeptin, an inhibitor of the lysosomal endopeptidase cathepsin, decreased the cytotoxicity of aMM46-MTX, supporting the involvement of lysosomal degradation of the conjugate in its action.
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Blair AH, Ghose TI (1983) Linkage of cytotoxic agents to immunoglobulins. J Immunol Methods 59:129
Bradford NM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248
Ehrlich P (1913) Chemotherapy. In: Himmelweit F (ed) (1960) The collected papers of Paul Ehrlich, vol III. Pergamon Press, London, p 510
Embleton MJ, Garnett MC (1985) Antibody targeting of anticancer agents. In: Baldwin RW, Byers VS (eds) Monoclonal antibodies for cancer detection and therapy. Academic Press, New York, p 317
Garnett MC, Baldwin RW (1986) An improved synthesis of a methotrexate-791T/36 monoclonal antibody conjugate cytotoxic to human osteogenic sarcoma cell lines. Cancer Res 46:2407
Garnett MC, Embleton MJ, Jacobs E, Baldwin RW (1983) Preparation and properties of a drug carrier antibody conjugate showing selective antibody-directed cytotoxicity in vitro. Int J Cancer 31:661
Ghose T, Blair AH (1978) Antibody-linked cytotoxic agents in the treatment of cancer: current status and future prospects. J Natl Cancer Inst 61:657
Halpern SE, Hagan PL, Garver PR, Koziol JA, Chen AWN, Frincke JM, Bartholomew RM, David GS, Adams TH (1983) Stability, characterization, and kinetics of 111In-labeled monoclonal antitumor antibodies in normal animals and nude mouse human tumor models. Cancer Res 43:5347
Huang A, Kennel SJ, Huang L (1983) Interactions of immunoliposomes with target cells. J Biol Chem 258:14034
Irie RF (1971) Antigenic cross-reactivity between primary spontaneous mouse mammary tumors and their transplantable ascites tumors. Cancer Res 31:1682
Irie RF, Nishioka K, Tachibana T, Takeuchi S (1969) Immunological studies on mouse mammary tumors. IV. Extraction and solubilization of transplantation antigen of mouse mammary tumor. Int J Cancer 4:150
Kanellos J, Pietersz GA, McKenzie FC (1985) Studies of methotrexate monoclonal antibody conjugates for immunotherapy. J Natl Cancer Inst 75:319
Kulkarni PN, Blair AH, Ghose TI (1981) Covalent binding of methotrexate to immunoglobulins and the effect of antibody-linked drug on tumor growth in vivo. Cancer Res 41:2700
Manabe Y, Tsubota T, Haruta Y, Kataoka K, Okazaki M, Haisa S, Nakamura K, Kimura I (1984) Production of a monoclonal antibody-methotrexate conjugate utilizing dextran T-40 and its biological activity. J Lab Clin Med 104:445
Order SE, Klein JL, Leichner PK (1981) Antiferritin IgG antibody for isotopic cancer therapy. Oncology 38:154
Royston I, Majda JA, Baird SM, Meserve BL, Griffiths JC (1979) Monoclonal antibody specific for human T lymphocytes: identification of normal and malignant T cells. Blood 54 (Suppl 1): 106A
Seto M, Umemoto N, Saito M, Masuho Y, Hara T, Takahashi T (1982) Monoclonal anti-MM46 antibody:ricin A chain conjugate: in vitro and in vivo antitumor activity. Cancer Res 42:5209
Seto M, Takahashi T, Tanimoto M, Nishizuka Y (1982) Production of monoclonal antibodies against MM antigens: the serologic identification of MM antigen with Ly-6.2 alloantigen. J Immunol 128:201
Seto M, Takahashi T, Nakamura S, Matsudaira Y, Nishizuka Y (1983) In vivo antitumor effects of monoclonal antibodies with different immunoglobulin classes. Cancer Res 43:4768
Uadia P, Blair AH, Ghose T (1983) Uptake of methotrexate linked to an anti-EL4-lymphoma antibody by EL4 cells. Cancer Immunol Immunother 16:127
Uadia P, Blair AH, Ghose T, Ferrone S (1985) Uptake of methotrexate linked to polyclonal and monoclonal antimelanoma antibodies by a human melanoma cell line. J Natl Cancer Inst 74:29
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Endo, N., Takeda, Y., Kishida, K. et al. Target-selective cytotoxicity of methotrexate conjugated with monoclonal anti-MM46 antibody. Cancer Immunol Immunother 25, 1–6 (1987). https://doi.org/10.1007/BF00199293
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DOI: https://doi.org/10.1007/BF00199293