Abstract
Immunotoxins (ITs) are chimeric proteins consisting of an antibody linked to a toxin. The antibody confers specificity (ability to recognize and react with the target), whereas the toxin confers cytotoxicity (ability to kill the target) (1–3). ITs have been used in both mice and humans to eliminate tumor cells, autoimmune cells, and virus-infected cells (4–6.
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References
Ghetie, V. and Vitetta, E. S. (1994) Immunotoxins in the therapy of cancer: from bench to clinic. Pharmacol. Ther. 63(3), 209–234.
Thrush, G. R., Lark, L. R., and Vitetta, E. S. (1996) Immunotoxins (review), in Therapeutic Immunology (Austen, K. F., Burakoff, S. J., Rosen, F. S., and Strom, T. B., eds.), Blackwell Science, Boston, pp. 385–397.
Pai, L. H. and Pastan, I. (1993) Immunotoxin therapy for cancer. JAMA 269, 78–81.
Frankel, A. E., Tagge, E. P., and Willingham, M. C. (1995) Clinical trials of targeted toxins. Semin. Cancer Biol. 6, 307–317.
Ghetie, M. A. and Vitetta, E. S. (1994) Recent developments in immunotoxin therapy. Curr. Opin. Immunol. 6, 707–714.
Grossbard, M. and Nadler, L. M. (1994) Immunotoxin therapy of lymphoid neoplasms. Semin. Hematol. 31, 88–97.
Wong, S. S. (1991) Chemistry of Protein Conjugation and Cross-Linking. CRC, Boca Raton, FL, pp. 267–294.
Vitetta, E. S., Thorpe, P. E., and Uhr, J. W. (1993) Immunotoxins: magic bullets or misguided missiles. Trends Pharmacol. Sci. 14, 148–154.
Brinkmann, U. and Pastan, I. (1994) Immunotoxins against cancer. Biochim. Biophys. Acta 1198, 27–45.
Kreitman, R. J. and Pastan, I. (1994) Recombinant toxins. Adv. Pharmacol. 28, 193–219.
Carlsson, J., Drevin, H., and Axen, R. (1978) Protein thiolation and reversible protein-protein conjugation N-succinimidyl 3-(2-pyridyldithio)propionate, anew heterobifunctional reagent. Biochem. J. 173(3), 723–737.
Brinkley, M. A. (1992) A survey of methods for preparing protein conjugates with dyes, haptens and crosslinking reagents. Bioconjug. Chem. 3, 2–13.
Thorpe, P. E., Wallace, P. M., Knowles, P. P., Relf, M. G., Brown, A. N. F., Watson, G. J., et al. (1988) Improved anti-tumor effects of immunotoxins prepared with deglycosylated ricin A chain and hindered disulfide linkages. Cancer Res. 48, 6396–6403.
FitzGerald, D., Idziorek, T., Batra, J. K., Willingham, M., and Pastan, I. (1990) Antitumor activity of a thioether-linked immunotoxin: OVB3-PE. Bioconjug. Chem. 1, 264–268.
Lambert, J. M., Goldmacher, V. S., Collinson, A. R., Nadler, L. M., and Blattler, W. A. (1991) An immunotoxin prepared with blocked ricin: a natural plant toxin adapted for therapeutic use. Cancer Res. 51, 6236–6242.
Harris, W. J. and Cunningham, C. (1995) Antibody Therapeutics. Landis, Austin, TX.
Goding, J. W. (1996) Monoclonal Antibodies: Principles and Practices. Academic, London, pp. 192–227.
Lamoyi, E. and Nisonoff, A. (1983) Preparation of F(ab’)2 fragments from mouse IgG of various subclasses. J. Immunol. Methods 50, 234–243.
Parham, P. (1983) On the fragmentation of monoclonal IgG1, IgG2a and IgG2b from BALB/c mice. J. Immunol. 131, 2895–2902.
Ghetie, V., Ghetie, M., Uhr, J. W., and Vitetta, E. S. (1988) Large scale preparation of immunotoxins constructed with the Fab’ fragment of IgG1 murine monoclonal antibodies and chemically deglycosylated ricin A chain. J. Immunol. Methods 112, 267–277.
Thorpe, P. E., Wallace, P. M., Knowles, P. P., Relf, M. G., Brown, A. N. F., Watson, G. J., et al. (1987) New coupling agents for the synthesis of immunotoxins containing a hindered disulfide bond with improved stability in vivo. Cancer Res. 47, 5924–5931.
Thorpe, P. E., Blakey, D. C., Brown, A. N., Knowles, P. P., Knyba, R. E., Wallace, P. M., et al. (1987) Comparison of two anti-Thy 1.1-abrin A-chain immunotoxins prepared with different cross-linking agents: antitumor effects, in vivo fate, and tumor cell mutants. J. Natl. Cancer Inst. 79, 1101–1112.
Ghetie, V., Thorpe, P. E., Ghetie, M., Knowles, P., Uhr, J. W., and Vitetta, E. S. (1991) The GLP large scale preparation of immunotoxins containing deglycosylated ricin A chain and a hindered disulfide bond. J. Immunol. Methods 142, 223–230.
Lambert, J. M., Blattler, W. A., McIntyre, G. D., Goldmacher, V. S., and Scott, C. F., Jr. (1988) Immunotoxins containing single chain ribosome-inactivating proteins, in Immunotoxins (Franker, A. E., ed.), Kluwer, Norwell, MA, pp. 175–213.
Duncan, R. J., Weston, P. D., and Wrigglesworth, R. (1983) A new reagent which may be used to introduce sulfhydryl groups into proteins, and its use in the preparation of conjugates for immunoassay. Anal. Biochem. 132, 68–73.
Hashida, S., Imagawa, M., Inque, S., Ruan, K. H., and Ishikawa, E. (1983) More useful maleimide compounds for the conjugation of Fab to horseradish peroxidase through thiol groups in the hinge. J. Appl. Biochem. 6, 56–63.
Lambert, J. M., McIntyre, G., Gauthier, M. N., Zullo, D., Rao, V., Steeves, R. M., et al. (1997) The galactose-binding sites of the cytotoxic lectin ricin can be chemically blocked in high yield with reactive ligands prepared by chemical modification of glycopeptides containing triantennary N-linked oligosaccharides. Biochemistry 30, 3234–3247.
Thorpe, P. E., Detre, S. I., Foxwell, B. M. J., Brown, A. N. F., Skilleter, D. N., Wilson, G., et al. (1985) Modification of the carbohydrate in ricin with metaperiodate-cyanoborohydride mixtures. Effects on toxicity and in vivo distribution. Eur. J. Biochem. 147, 197–206.
Ghetie, M., May, R. D., Till, M., Uhr, J. W., Ghetie, V., Knowles, P. P., et al. (1988) Evaluation of ricin A chain-containing immunotoxins directed against CD 19 and CD22 antigens on normal and malignant human B-cells as potential reagents for in vivo therapy. Cancer Res. 48, 2610–2617.
Ghetie, V., Till, M. A., Ghetie, M., Tucker, T., Porter, J., Patzer, E. J., et al. (1990) Preparation and characterization of conjugates of recombinant CD4 and deglycosylated ricin A chain using different cross-linkers. Bioconjug Chem. 1, 24–31.
Fulton, R. J., Blakey, D. C., Knowles, P. P., Uhr, J. W., Thorpe, P. E., and Vitetta, E. S. (1986) Production of ricin A1, A2, and B chains and characterization of their toxicity. J. Biol. Chem. 261, 5314–5319.
Irvin, J. D. (1983) Pokeweed antiviral protein. Pharmacol. Ther. 21, 371–387.
Irvin, J. D. and Uckun, F. M. (1997) Pokeweed antiviral protein: Ribosome inactivation and therapeutic applications. Pharmacol. Ther. 55, 279–302.
Myers, D. E., Irvin, J. D., Smith, R. S., Kuebelbeck, V. M., and Uckun, F. M. (1991) Production of a pokeweed antiviral protein (PAP)-containing immunotoxin, B43-PAP, directed against the CD19 human B lineage lymphoid differentiation antigen in highly purified form for human clinical trials. J. Immunol. Methods 136, 221–237.
Ogata, M., Chaudhary, V. K., Pastan, I., and Fitzgerald, D. J. (1997) Processing of pseudomonas exotoxin by a cellular protease results in the generation of a 37,000-Da toxin fragment that is translocated to the cytosol. J. Biol. Chem. 265, 20, 678–20,685.
Allured, V. S., Collier, R. J., Carroll, S. F., and McKay, D. B. (1986) Structure of exotoxin A of Pseudomonas aeruginosa at 3.0-Angstrom resolution. Proc. Natl. Acad. Sci. USA 83, 1320–1324.
Hwang, J., FitzGerald, D. J., Adhya, S., and Pastan, I. (1987) Functional domains of Pseudomonas exotoxin identified by deletion analysis of the gene expressed in E. coli. Cell 48, 129–136.
Kreitman, R. J., Hansen, H. J., Jones, A. L., FitzGerald, D. J. P., Goldenberg, D. M., and Pastan, I. (1993) Pseudomonas exotoxin-based immunotoxins containing the antibody LL2 or LL2-Fab’ induce regression of subcutaneous human B-cell lymphoma in mice. Cancer Res. 53, 819–825.
Mansfield, E., Pastan, I., FitzGerald, D. J. (1996) Characterization of RFB4-Pseudomonas exotoxin A immunotoxins targeted to CD22 on B-cell malignancies. Bioconjug. Chem. 7, 557–563.
Ghetie, V., Swindell, E., Uhr, J. W., and Vitetta, E. S. (1993) Purification and properties of immunotoxins containing one vs two deglycosylated ricin A chain. J. Immunol. Methods 166, 117–122.
Ghetie, V., Engert, A., Schnell, R., and Vitetta, E. S. (1995) The in vivo anti-tumor activity of immunotoxins containing two vs one deglycosylated ricin A chains. Cancer Lett. 98(1), 97–101.
Fraker, P. J. and Speck, J. C., Jr. (1978) Protein and cell membrane iodinations with a sparingly soluble chloramide, 1,3,4,6-tetrachloro-3a,6a-diphenylglycoluril. Biochem. Biophys. Res. Commun. 80, 849–857.
Ghetie, M. A., Richardson, J., Tucker, T., Jones, D., Uhr, J. W., and Vitetta, E. S. 1990) Disseminated or localized growth of a human B-cell tumor (Daudi) in SCID mice. Int. Cancer 45, 481–485.
Ghetie, M. A., Richardson, J., Tucker, T., Jones, D., Uhr, J. W., and Vitetta, E. S. (1991) Antitumor activity of Fab’ and IgG-anti-CD22 immunotoxins in disseminated human B lymphomas grown in mice with severe combined immunodeficiency disease: effect on tumor cells in extranodal sites. Cancer Res. 51, 5876–5880.
Ghetie, M. A., Tucker, K., Richardson, J., Uhr, J. W., and Vitetta, E. S. (1992) The antitumor activity of an anti-CD22 immunotoxin in SCID mice with disseminated Daudi lymphoma is enhanced by either an anti-CD 19 antibody or an anti-CD 19 immunotoxin. Blood 80, 2315–2320.
Uckun, F. M., Ramakrishnan, S., and Houston, L. L. (1985) Immunotoxin-mediated elimination of clonogenic tumor cells in the presence of human bone marrow. J. Immunol. 134, 2010–2016.
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Ghetie, V., Vitetta, E.S. (2000). Chemical Construction of Immunotoxins. In: Francis, G.E., Delgado, C. (eds) Drug Targeting. Methods in Molecular Medicine™, vol 25. Humana Press. https://doi.org/10.1385/1-59259-075-6:1
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DOI: https://doi.org/10.1385/1-59259-075-6:1
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