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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1980 Aug;77(8):4539–4543. doi: 10.1073/pnas.77.8.4539

Antibody-directed cytotoxic agents: use of monoclonal antibody to direct the action of toxin A chains to colorectal carcinoma cells.

D G Gilliland, Z Steplewski, R J Collier, K F Mitchell, T H Chang, H Koprowski
PMCID: PMC349879  PMID: 6933502

Abstract

We have constructed cell-specific cytotoxic agens by covalently coupling the A chain from diphtheria toxin or ricin toxin to monoclonal antibody directed against a colorectal carcinoma tumor-associated antigen. Antibody 1083-17-1A was modified by attachment of 3-(2-pyridyldithio)propionyl or cystaminyl groups and then treated with reduced A chain to give disulfide-linked conjugates that retained the original binding specificity of the antibody moiety. the conjugates showed cytotoxic activity for colorectal carcinoma cells in culture, but were not toxic in the same concentration range for a variety of cell lines that lacked the antigen. Under defined conditions virtually 100% of antigen-bearing cultured cells were killed, whereas cells that lacked the antigen were not affected. Conjugates containing toxin A chains coupled to monoclonal antibodies may be useful in studying functions of various cell surface components and, possibly, as tumor-specific therapeutic agents.

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Selected References

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  1. Benson S., Olsnes S., Pihl A., Skorve J., Abraham A. K. On the mechanism of protein-synthesis inhibition by abrin and ricin. Inhibition of the GTP-hydrolysis site on the 60-S ribosomal subunit. Eur J Biochem. 1975 Nov 15;59(2):573–580. doi: 10.1111/j.1432-1033.1975.tb02484.x. [DOI] [PubMed] [Google Scholar]
  2. Boquet P., Silverman M. S., Pappenheimer A. M., Jr, Vernon W. B. Binding of triton X-100 to diphtheria toxin, crossreacting material 45, and their fragments. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4449–4453. doi: 10.1073/pnas.73.12.4449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burstein S., Knapp R. Chemotherapy of murine ovarian carcinoma by methotrexate-antibody conjugates. J Med Chem. 1977 Jul;20(7):950–952. doi: 10.1021/jm00217a018. [DOI] [PubMed] [Google Scholar]
  4. Cawley D. B., Hedblom M. L., Houston L. L. Homology between ricin and Ricinus communis agglutinin: amino terminal sequence analysis and protein synthesis inhibition studies. Arch Biochem Biophys. 1978 Oct;190(2):744–755. doi: 10.1016/0003-9861(78)90335-1. [DOI] [PubMed] [Google Scholar]
  5. Chung D. W., Collier R. J. Enzymatically active peptide from the adenosine diphosphate-ribosylating toxin of Pseudomonas aeruginosa. Infect Immun. 1977 Jun;16(3):832–841. doi: 10.1128/iai.16.3.832-841.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chung D. W., Collier R. J. The mechanism of ADP-ribosylation of elongation factor 2 catalyzed by fragment A from diphtheria toxin. Biochim Biophys Acta. 1977 Aug 11;483(2):248–257. doi: 10.1016/0005-2744(77)90053-5. [DOI] [PubMed] [Google Scholar]
  7. Collier R. J. Diphtheria toxin: mode of action and structure. Bacteriol Rev. 1975 Mar;39(1):54–85. doi: 10.1128/br.39.1.54-85.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Collier R. J., Kandel J. Structure and activity of diphtheria toxin. I. Thiol-dependent dissociation of a fraction of toxin into enzymically active and inactive fragments. J Biol Chem. 1971 Mar 10;246(5):1496–1503. [PubMed] [Google Scholar]
  9. Drazin R., Kandel J., Collier R. J. Structure and activity of diphtheria toxin. II. Attack by trypsin at a specific site within the intact toxin molecule. J Biol Chem. 1971 Mar 10;246(5):1504–1510. [PubMed] [Google Scholar]
  10. Flickinger R. A., Trost S. R. Cytotoxicity of antibody-phospholipase C conjugates on cultured Friend leukemia cells. Eur J Cancer. 1976 Feb;12(2):159–160. doi: 10.1016/0014-2964(76)90220-6. [DOI] [PubMed] [Google Scholar]
  11. Gerhard W., Croce C. M., Lopes D., Koprowski H. Repertoire of antiviral antibodies expressed by somatic cell hybrids. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1510–1514. doi: 10.1073/pnas.75.3.1510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ghose T., Guclu A., Tai J., MacDonald A. S., Norvell S. T., Aquino J. Antibody as carrier of 131I in cancer diagnosis and treatment. Cancer. 1975 Nov;36(5):1646–1657. doi: 10.1002/1097-0142(197511)36:5<1646::aid-cncr2820360518>3.0.co;2-a. [DOI] [PubMed] [Google Scholar]
  13. Ghose T., Nigam S. P. Antibody as carrier of chlorambucil. Cancer. 1972 May;29(5):1398–1400. doi: 10.1002/1097-0142(197205)29:5<1398::aid-cncr2820290542>3.0.co;2-d. [DOI] [PubMed] [Google Scholar]
  14. Ghose T., Norvell S. T., Guclu A., Cameron D., Bodurtha A., MacDonald A. S. Immunochemotherapy of cancer with chlorambucil-carrying antibody. Br Med J. 1972 Aug 26;3(5825):495–499. doi: 10.1136/bmj.3.5825.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gilliland D. G., Collier R. J., Moehring J. M., Moehring T. J. Chimeric toxins: toxic, disulfide-linked conjugate of concanavalin A with fragment A from diphtheria toxin. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5319–5323. doi: 10.1073/pnas.75.11.5319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Herlyn D., Herlyn M., Steplewski Z., Koprowski H. Monoclonal antibodies in cell-mediated cytotoxicity against human melanoma and colorectal carcinoma. Eur J Immunol. 1979 Aug;9(8):657–659. doi: 10.1002/eji.1830090817. [DOI] [PubMed] [Google Scholar]
  17. Herlyn M., Steplewski Z., Herlyn D., Koprowski H. Colorectal carcinoma-specific antigen: detection by means of monoclonal antibodies. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1438–1442. doi: 10.1073/pnas.76.3.1438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hurwitz E., Levy R., Maron R., Wilchek M., Arnon R., Sela M. The covalent binding of daunomycin and adriamycin to antibodies, with retention of both drug and antibody activities. Cancer Res. 1975 May;35(5):1175–1181. [PubMed] [Google Scholar]
  19. Koprowski H., Gerhard W., Croce C. M. Production of antibodies against influenza virus by somatic cell hybrids between mouse myeloma and primed spleen cells. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2985–2988. doi: 10.1073/pnas.74.7.2985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Koprowski H., Steplewski Z., Herlyn D., Herlyn M. Study of antibodies against human melanoma produced by somatic cell hybrids. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3405–3409. doi: 10.1073/pnas.75.7.3405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Koprowski H., Steplewski Z., Mitchell K., Herlyn M., Herlyn D., Fuhrer P. Colorectal carcinoma antigens detected by hybridoma antibodies. Somatic Cell Genet. 1979 Nov;5(6):957–971. doi: 10.1007/BF01542654. [DOI] [PubMed] [Google Scholar]
  22. Levy R., Hurwitz E., Maron R., Arnon R., Sela M. The specific cytotoxic effects of daunomycin conjugated to antitumor antibodies. Cancer Res. 1975 May;35(5):1182–1186. [PubMed] [Google Scholar]
  23. Masuho Y., Hara T., Noguchi T. Preparation of a hybrid of fragment Fab' of antibody and fragment A of diphtheria toxin and its cytotoxicity. Biochem Biophys Res Commun. 1979 Sep 12;90(1):320–326. doi: 10.1016/0006-291x(79)91627-9. [DOI] [PubMed] [Google Scholar]
  24. Miskimins W. K., Shimizu N. Synthesis of a cytotoxic insulin cross-linked to diphtheria toxin fragment A capable of recognizing insulin receptors. Biochem Biophys Res Commun. 1979 Nov 14;91(1):143–151. doi: 10.1016/0006-291x(79)90595-3. [DOI] [PubMed] [Google Scholar]
  25. Moolten F. L., Capparell N. J., Cooperband S. R. Antitumor effects of antibody-diphtheria toxin conjugates: use of hapten-coated tumor cells as an antigenic target. J Natl Cancer Inst. 1972 Oct;49(4):1057–1062. [PubMed] [Google Scholar]
  26. Moolten F. L., Capparell N. J., Zajdel S. H., Cooperband S. R. Antitumor effects of antibody-diphtheria toxin conjugates. II. Immunotherapy with conjugates directed against tumor antigens induced by simian virus 40. J Natl Cancer Inst. 1975 Aug;55(2):473–477. [PubMed] [Google Scholar]
  27. Nicolson G. L., Blaustein J. The interaction of Ricinus communis agglutinin with normal and tumor cell surfaces. Biochim Biophys Acta. 1972 May 9;266(2):543–547. doi: 10.1016/0005-2736(72)90109-5. [DOI] [PubMed] [Google Scholar]
  28. Oeltmann T. N., Heath E. C. A hybrid protein containing the toxic subunit of ricin and the cell-specific subunit of human chorionic gonadotropin. I. Synthesis and characterization. J Biol Chem. 1979 Feb 25;254(4):1022–1027. [PubMed] [Google Scholar]
  29. Oeltmann T. N., Heath E. C. A hybrid protein containing the toxic subunit of ricin and the cell-specific subunit of human chorionic gonadotropin. II. Biologic properties. J Biol Chem. 1979 Feb 25;254(4):1028–1032. [PubMed] [Google Scholar]
  30. Olsnes S., Pihl A. Different biological properties of the two constituent peptide chains of ricin, a toxic protein inhibiting protein synthesis. Biochemistry. 1973 Jul 31;12(16):3121–3126. doi: 10.1021/bi00740a028. [DOI] [PubMed] [Google Scholar]
  31. Rowland G. F., O'Neill G. J., Davies D. A. Suppression of tumour growth in mice by a drug-antibody conjugate using a novel approach to linkage. Nature. 1975 Jun 5;255(5508):487–488. doi: 10.1038/255487a0. [DOI] [PubMed] [Google Scholar]
  32. Shearer W. T., Turnbaugh T. R., Coleman W. E., Aach R. D., Philpott G. W., Parker C. W. Cytotoxicity with antibody-glucose oxidase conjugates specific for a human colonic cancer and carcinoembryonic antigen. Int J Cancer. 1974 Oct 15;14(4):539–547. doi: 10.1002/ijc.2910140414. [DOI] [PubMed] [Google Scholar]
  33. Stuchbury T., Shipton M., Norris R., Malthouse J. P., Brocklehurst K., Herbert J. A., Suschitzky H. A reporter group delivery system with both absolute and selective specificity for thiol groups and an improved fluorescent probe containing the 7-nitrobenzo-2-oxa-1,3-diazole moiety. Biochem J. 1975 Nov;151(2):417–432. doi: 10.1042/bj1510417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thorpe P. E., Ross W. C., Cumber A. J., Hinson C. A., Edwards D. C., Davies A. J. Toxicity of diphtheria toxin for lymphoblastoid cells is increased by conjugation to antilymphocytic globulin. Nature. 1978 Feb 23;271(5647):752–755. doi: 10.1038/271752a0. [DOI] [PubMed] [Google Scholar]
  35. Trouet A. Perspectives in cancer research. Increased selectivity of drugs by linking to carriers. Eur J Cancer. 1978 Feb;14(2):105–111. doi: 10.1016/0014-2964(78)90167-6. [DOI] [PubMed] [Google Scholar]
  36. Uchida T., Yamaizumi M., Mekada E., Okada Y., Tsuda M., Kurokawa T., Sugino Y. Reconstitution of hybrid toxin from Fragment A of diphtheria toxin and a subunit of Wistaria floribunda lectin. J Biol Chem. 1978 Sep 25;253(18):6307–6310. [PubMed] [Google Scholar]
  37. Wiktor T. J., Koprowski H. Monoclonal antibodies against rabies virus produced by somatic cell hybridization: detection of antigenic variants. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3938–3942. doi: 10.1073/pnas.75.8.3938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yamaguchi T., Kato R., Beppu M., Terao T., Inoue Y., Ikawa Y., Osawa T. Preparation of concanavalin A-ricin A-chain conjugate and its biologic activity against various cultured cells. J Natl Cancer Inst. 1979 Jun;62(6):1387–1395. [PubMed] [Google Scholar]

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