Abstract
Immunotoxins were prepared with a Ber-H2 (anti-CD30) monoclonal antibody and native or recombinant dianthin 30, a ribosome-inactivating protein fromDianthus caryophyllus (carnation). Both immunotoxins selectively inhibited protein synthesis by CD30+ cell lines D430B (lymphoblastoid, infected with Epstein-Barr virus). L428 and L540 (both from Hodgkin's lymphoma). IC50 values (concentrations, as dianthin, causing 50% inhibition) ranged from 324 pM to 479 pM (immunotoxin with native dianthin 30) or from 45 pM to 182 pM (immunotoxin with recombinant dianthin 30). The effect of either immunotoxin on protein synthesis by the CD30− cell line K562 (from a chronic myeloid leukaemia) was not different from that of free dianthin (IC50 higher than nM).
Key words: Immunotoxin, Dianthin, Ribosome-inactivating protein, Hodgkin's lymphoma
References
- 1.Barbieri L, Battelli MG, Stirpe F. Ribosome-inactivating proteins from plants. Biochim Biophys Acta. 1993;1154:327. doi: 10.1016/0304-4157(93)90002-6. [DOI] [PubMed] [Google Scholar]
- 2.Endo Y. Mechanism of action of ricin and related toxins on the inactivation of ribosomes. In: Frankel AE, editor. Immunotoxins. Boston: Kluwer; 1988. p. 75. [DOI] [PubMed] [Google Scholar]
- 3.Ramakrishnan S, Fryxell D, Mohanraj D, Olson M, Li BY. Cytotoxic conjugates containing translational inhibitory proteins. Annu Rev Pharmacol Toxicol. 1992;32:579. doi: 10.1146/annurev.pa.32.040192.003051. [DOI] [PubMed] [Google Scholar]
- 4.Vitetta ES, Thorpe PE, Uhr JW. Immunotoxins: magic bullets or misguided missiles? Immunol Today. 1993;14:252. doi: 10.1016/0167-5699(93)90041-I. [DOI] [PubMed] [Google Scholar]
- 5.Pastan I, Chaudhary V, FitzGerald D. Recombinant toxins as novel therapeutic agents. Annu Rev Biochem. 1992;61:331. doi: 10.1146/annurev.bi.61.070192.001555. [DOI] [PubMed] [Google Scholar]
- 6.Legname G, Bellosta P, Gromo G, Modena D, Keen JN, Roberts LM, Lord JM. Nucleotide sequence of cDNA coding for dianthin-30, a ribosome-inactivating protein fromDianthus caryophyllus . Biochim Biophys Acta. 1991;1090:119. doi: 10.1016/0167-4781(91)90046-o. [DOI] [PubMed] [Google Scholar]
- 7.Legname G, Gromo G, Lord JM, Monzini N, Lord JM. Expression and activity of pre-dianthin 30 and dianthin 30. Biochem Biophys Res Commun. 1993;192:1230. doi: 10.1006/bbrc.1993.1548. [DOI] [PubMed] [Google Scholar]
- 8.Stirpe F, Williams DG, Onyon LJ, Legg RF, Stevens WA. Dianthins, ribosome-damaging proteins with anti-viral properties fromDianthus caryophyllus L. (carnation) Biochem J. 1981;195:399. doi: 10.1042/bj1950399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 9.Falini BA, Bolognesi A, Flenghi L, Tazzari PL, Broe MK, Stein H, Dürkop H, Aversa F, Corneli P, Pizzolo G, Barbabietola G, Sabattini E, Pileri S, Martelli MF, Stirpe F. Response of refractory Hodgkin's disease to monoclonal anti-CD30 immunotoxin. Lancet. 1992;339:1195. doi: 10.1016/0140-6736(92)91135-u. [DOI] [PubMed] [Google Scholar]
- 10.Barbieri L, Stoppa C, Bolognesi A. Large scale chromatographic purification of ribosome-inactivating proteins. J Chromatogr. 1987;408:235. [Google Scholar]
- 11.Sambrook J, Fritsch EF, Maniatis T. Molecular cloning, a laboratory manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press; 1989. [Google Scholar]
- 12.Studier FW, Rosemberg AH, Dunn JJ, Dubendorff JW. Methods Enzymol. 1990;185:61. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
- 13.Schwarting R, Gerde J, Dürkop H, Falini B, Pileri S, Stein H. Ber-H2: a new anti-K1 (CD30) monoclonal antibody directed at a formol-resistant epitope. Blood. 1989;74:1678. [PubMed] [Google Scholar]
- 14.Bolognesi A, Barbieri L, Carnicelli D, Abbondanza A, Cenini P, Falasca AI, Dinota A, Stirpe F. Purification and properties of a new ribosome-inactivating protein with RNAN-glycosidase activity suitable for immunotoxin preparation from the seeds ofMomordica cochinchinensis . Biochim Biophys Acta. 1989;993:287. doi: 10.1016/0304-4165(89)90178-5. [DOI] [PubMed] [Google Scholar]
- 15.Poccia F, Piselli P, Di Cesare F, Bach S, Colizzi V, Mattei M, Bolognesi A, Stirpe F. Recognition and killing of tumour expressing heat shock protein 65 kD with immunotoxins containing saporin. Br J Cancer. 1992;66:427. doi: 10.1038/bjc.1992.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16.Ellman GL. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959;82:70. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
- 17.Bolognesi A, Tazzari PL, Tassi C, Gromo G, Gobbi M, Stirpe F. Comparison of anti-lymphocyte immunotoxins containing different ribosome-inactivating proteins and antibodies. Clin Exp Immunol. 1992;89:341. doi: 10.1111/j.1365-2249.1992.tb06959.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 18.Schaadt M, Diehl V, Stein H, Fonatsch H, Kirchner H. Two neoplastic cell lines with unique features derived from Hodgkin's disease. Int J Cancer. 1980;26:723. doi: 10.1002/ijc.2910260605. [DOI] [PubMed] [Google Scholar]
- 19.Engert A, Burrows E, Jung W, Tazzari PL, Stein H, Pfreundschuh M, Diehl V, Thorpe P. Evaluation of ricin A chain-containing immunotoxins directed against the CD30 antigen as potential reagent for the treatment of Hodgkin's disease. Cancer Res. 1990;50:84. [PubMed] [Google Scholar]
- 20.Ferreras JM, Barbieri L, Girbés T, Battelli MG, Rojo MA, Arias FJ, Rocher MA, Soriano F, Mendez E, Stirpe F. Distribution and properties of major ribosome-inactivating proteins (28 S rRNAN-glycosidases) of the plantSaponaria officinalis L. (Caryophyllaceae) Biochim Biophys Acta. 1993;1216:31. doi: 10.1016/0167-4781(93)90034-b. [DOI] [PubMed] [Google Scholar]
- 21.Tazzari PL, Bolognesi A, De Totero D, Falini B, Lemoli RM, Soria MR, Pileri S, Gobbi M, Stein H, Flenghi L, Martelli MF, Stirpe F. Ber-H2 (anti-CD30)-saporin immunotoxin: a new tool for the treatment of Hodgkin's disease and CD30+ lymphoma. In vitro evaluation. Br J Haematol. 1992;81:203. doi: 10.1111/j.1365-2141.1992.tb08208.x. [DOI] [PubMed] [Google Scholar]
- 22.Benatti L, Nitti G, Solinas M, Valsasina B, Vitale A, Ceriotti A, Soria MR. A saporin-6 cDNA containing a precursor sequence coding for a carboxyl-terminal extension. FEBS Lett. 1991;291:285. doi: 10.1016/0014-5793(91)81303-p. [DOI] [PubMed] [Google Scholar]
- 23.Wawrzinczak EJ, Cumber AJ, Henry RV, Parnell GD, Westwood JH. Structural factors influencing the pharmacokinetics and stability of immunotoxins. Biochem Soc Trans. 1992;20:738. doi: 10.1042/bst0200738. [DOI] [PubMed] [Google Scholar]
- 24.Duke-Cohan JS, Morimoto C, Schlossman SF. Depletion of the helper/inducer (memory) T cell subset using a bispecific antibody-toxin conjugate directed against CD4 and CD29. Transplantation. 1993;56:1188. doi: 10.1097/00007890-199311000-00027. [DOI] [PubMed] [Google Scholar]