Summary
In studies of antitumor antibody-cytotoxic agent conjugates as potential antitumor agents with improved tumor specificity, the toxic subunit A-chain of ricin was conjugated with a monoclonal antibody to a tumor-associated antigen expressed weakly on murine leukemia L1210 cells and strongly on L1210/GZL cells, a guanazole-resistant subline of L1210, employing N-succinimidyl 3-(2-pyridyldithio)propionate as cross-linking agent. The conjugate (anti-L1210 conjugate) exhibited a potent concentration-dependent cytotoxicity against cultured L1210/GZL cells, and inhibited cell growth at concentrations over 0.8 μg/ml. The conjugate killed all L1210/GZL cells at a concentration of 100 μg/ml. Neither nonimmune conjugate similarly prepared from mouse nonimmune IgG nor unconjugated anti-L1210 IgG alone showed cytotoxicity against L1210/GZL cells. When (BALB/c×DBA/2)F1 mice inoculated with 1 × 105 L1210/GZL cells were treated with IP injections of 27 μg anti-L1210 conjugate 1 h and 5 days after tumor cell inoculation, a life-prolonging effect was observed. [Lifespan in treated animals as percentage of that in controls (T/C)=146%]. However, when the dose per injection was increased to 50 μg per mouse, survival was the same as in the control group. Postmortem examination of mice that had been treated with 50 μg anti-L1210 conjugate revealed lesions with necrosis and hemorrhage in the liver parenchyma and the intestinal epithelium, respectively. A similar toxic effect on the host mice was also observed with nonimmune conjugate.
Similar content being viewed by others
References
Blythman HE, Cassellas P, Gros O, Gros P, Jansen FK, Paolucci F, Pau B, Vidal H (1981) Immunotoxins: hybrid molecules of monoclonal antibodies and a toxin subunit specifically kill tumor cells. Nature 290:145
Carlsson J, Drevin H, Axen R (1978) Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. Biochem J 173:723
Ey PL, Prowse SJ, Jenkin CR (1978) Isolation of pure IgG1, IgG2a, and IgG2b immunoglobulins from mouse serum using protein A-Sepharose. Immunochemistry 15:429
Fuji H (1981) Differential recognition of tumor cell variants by B and T lymphocytes. In: Steinberg C, Lefkovitz I (eds) The immune system, vol 2. Karger, Basel, p 431
Fuji H, Mihich E, Pressman D (1977) Differential tumor immunogenicity of L1210 and its sublines. I. Effect of an increased antigen density on tumor cell surfaces on primary B cell responses in vitro. J Immunol 119:983
Fuji H, Mihich E, Pressman D (1979) Differential tumor immunogenicity of DBA/2 mouse lymphoma L1210 and its sublines. II. Increased expression of tumor-associated antigens on subline cells recognized by serologic and transplantation methods. J Natl Cancer Inst 62:1503
Gilliland DG, Steplewski Z, Collier RJ, Mitchell KF, Chang TH, Koprowski H (1980) Antibody-directed cytotoxic agents: use of monoclonal antibody to direct the action of toxin A chains to colorectal carcinoma cells. Proc Natl Acad Sci USA 77:4539
Jansen FK, Blythman HE, Carriere D, Casellas P, Diaz J, Gros P, Hennequin JR, Paolucci F, Pau B, Poncelet P, Richer G, Salhi SL, Vidal H, Voisin GA (1981) High specific cytotoxicity of antibody-toxin hybrid molecules (immunotoxins) for target cells. Immunol Lett 2:97
Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495
Krolick KA, Villemez C, Isakson P, Uhr JW, Vitetta ES (1980) Selective killing of normal or neoplastic B cells by antibodies coupled to the A chain of ricin. Proc Natl Acad Sci USA 77:5419
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
Masuho Y, Hara T (1980) Target-cell cytotoxicity of a hybrid of Fab' of immunoglobulin and A-chain of ricin. Gann 71:759
Masuho Y, Hara T, Noguchi T (1979) Preparation of a hybrid of fragment Fab' of antibody and fragment A of diphtheria toxin and its cytotoxicity. Biochem Biophys Res Commun 90:320
Oeltmann TN, Forbes JT (1981) Inhibition of mouse spleen cell function by diphtheria toxin fragment A coupled to anti-mouse Thy-1.2 and by ricin A-chain coupled to anti-mouse IgM. Arch Biochem Biophys 209:362
Olsnes S, Pihl A (1973) Different biological properties of the two constituent peptide chains of ricin, a toxic protein inhibiting protein synthesis. Biochemistry 12:3121
Rapp L, Fuji H (1983) Differential antigenic expression of the DBA/2 lymphoma L1210 and its sublines. Cross-reactivity with C3H mammary tumors as defined by syngeneic monoclonal antibodies. Cancer Res 43:2592
Raso V, Griffin T (1980) Specific cytotoxicity of a human immunoglobulin-directed Fab'-ricin A chain conjugate. J Immunol 125:2610
Raso V, Ritz J, Basala M, Schlossman SF (1982) Monoclonal antibody-ricin A-chain conjugate selectively cytotoxic for cells bearing the common acute lymphoblastic leukemia antigen. Cancer Res 42:457
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
Tsukada Y, Bischof WK-D, Hibi N, Hirai H, Hurwitz E, Sela M (1982) Effect of a conjugate of daunomycin and antibodies to rat α-fetoprotein on the growth of α-fetoprotein-producing tumor cells. Proc Natl Acad Sci USA 79:621
Weber K, Osborn M (1969) The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem 244:4406
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kishida, K., Masuho, Y., Saito, M. et al. Ricin A-chain conjugated with monoclonal anti-L1210 antibody. Cancer Immunol Immunother 16, 93–97 (1983). https://doi.org/10.1007/BF00199238
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00199238