Abstract
Cell targeting agents such as antibodies, antibody fragments (sFvs), or growth factors have been conjugated or genetically fused to a variety of plant and bacterial toxins. These targeted therapeutics, termed “immunotoxins,” have been evaluated for their clinical efficacy in the treatment of cancer, AIDS, and immunological diseases (1,2). Development of potentially promising clinical results, however, have been hampered by problems of toxicity and immunogenicity owing to the foreign proteins (3–8). Although the development of humanized antibodies has alleviated some of these effects (9,10), the toxins still remain a problem. In this regard the use of human proteins as components of the immunotoxin are highly desirable (reviewed in ref. 11). Human RNases such as EDN, angiogenin and pancreatic RNase A are not toxic to cells yet when linked chemically or fused genetically to cell surfacebinding ligands have potent anti-tumor effects both in vitro and in vivo (12–23). Furthermore in vivo experiments demonstrate that the RNase-based therapeutics cause fewer nonspecific toxic or immunogenic side effects than plant and bacterial toxins (22,23 and reviewed in ref. 11).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Vitetta E. S., Thorpe P. E., and Uhr J. W. (1993). Immunotoxins: magic bullets or misguided missiles. TiPS 14, 148–154.
Pastan I. (1997). Targeted therapy of cancer with recombinant immunotoxins. Biochim. Biophys. Acta 1333, C1–C6.
Sawler D. L., Bartholomew R. M., Smith L. M., and Dillman R. (1985). Human immune response to multiple injections of murine monoclonal IgG. J. Immunol. 135, 1530–1535.
Schroff R. W., Foon K. A., Beatty S. M., Oldham R., and Morgan A. (1985). Human anti-murine immunoglobulin response in patients receiving monoclonal antibody therapy. Cancer Res. 45, 879–885.
Harkonen S., Stoudemire J., Mischak R., Spitler L., Lopez H., and Scannon P. (1987). Toxicity and immunogenicity of monoclonal antimelanoma antibody-ricin A chain immunotoxins in rats. Cancer Res. 47, 1377–1385.
Rybak S. M. and Youle R. J. (1991). Clinical use of immunotoxins: monoclonal antibodies conjugated to protein toxins. Immunol. Allergy Clin. N. Am. 11, 359–380.
Soler-Rodriguez A. M., Ghetie M.-A., Oppenheimer-Marks N., Uhr J. W., and Vitetta E. S. (1993). Ricin A-chain and ricin A-chain immunotoxins rapidly damage human endothelial cells: Implications for vascular leak syndrome. Exp. Cell Res. 206, 227–234.
Thrush G. R., Lark L. R., Clinchy B. C., and Vitetta E. S. (1996). Immunotoxins: An Update. Ann. Rev. Immunol. 14, 49–71.
Khazaeli M. B., Conry R. M., and LoBuglio A. F. (1994). Human immune response to monoclonal antibodies. J. Immunother. 15, 42–52.
Stephens S., Emtage S., Vetterlein O., Chaplin L., Bebbington C., Nesbitt A., et al. (1995). Comprehensive pharmacokinetics of a humanized antibody and analysis of residual anti-idiotypic responses. Immunology 85, 668–674.
Rybak S. M. and Newton D. L. (1999). Immunoenzymes, in Antibody Fusion Proteins (Chamow S. M. and Ashkenazi A., eds.), John Wiley & Sons, New York, NY, pp. 53–110.
Rybak S. M., Saxena S. K., Ackerman E. J., and Youle R. J. (1991). Cytotoxic potential of ribonuclease and ribonuclease hybrid proteins. J. Biol. Chem. 266, 21,202–21,207.
Newton D. L., Ilercil O., Laske D. W., Oldfield E., Rybak S. M., and Youle R. J. (1992). Cytotoxic ribonuclease chimeras: Targeted tumoricidal activity in vitro and in vivo. J. Biol. Chem. 267, 19,572–19,578.
Jinno H., Ueda M., Ozawa S., Kikuchi K., Ikeda T., Enomoto K., and Kitajima M. (1996). Epidermal growth factor receptor-dependent cytotoxic effect by an EGF-ribonuclease conjugate on human cancer cell lines: a trial for less immunogenic chimeric toxin. Can. Chemother. Pharmacol. 38, 303–308.
Newton D. L., Nicholls P. J., Rybak S. M., and Youle R. J. (1994). Expression and characterization of recombinant human eosinophil-derived neurotoxin and eosinophilderived neurotoxin-anti-transferrin receptor sFv. J. Biol. Chem. 269, 26,739–26,745.
Zewe M., Rybak S. M., Dubel S., Coy J. F., Welschof M., Newton D. L., and Little M. (1997). Cloning and cytotoxicity of a human pancreatic RNase immunofusion. Immunotechnology 3, 127–136.
Psarras K., Ueda M., Yamamura T., Ozawa S., Kitajima M., Aiso S., et al. (1998). Human pancreatic RNase1-human epidermal growth factor fusion: an entirely human immunotoxin analog with cytotoxic properties against squamous cell carcinomas. Prot. Eng. 11, 1285–1292.
Yoon J. M., Han S. H., Kown O. B., Kim S. H., Park M. H., and Kim B. K. (1999). Cloning and cytotoxicity of fusion proteins of EGF and angiogenin. Life Sci. 64, 1435–1445.
Newton D. L., Xue Y., Olson K. A., Fett J. W., and Rybak S. M. (1996). Angiogenin single-chain immunofusions: Influence of peptide linkers and spacers between fusion protein domains. Biochemistry 35, 545–553.
Rybak S. M., Hoogenboom H. R., Meade H. M., Raus J. C., Schwartz D., and Youle R. J. (1992). Humanization of immuntoxins. Proc. Natl. Acad. Sci. USA 89, 3165–3169.
Futami J., Seno M., Ueda M., Tada H., and Yamada H. (1999). Inhibition of cell growth by a fused protein of human ribonuclease 1 and human basic fibroblast growth factor. Prot. Eng. 12, 1013–1019.
Newton D. L., Pollock D., DiTullio P., Echelard Y., Harvey M., Wilburn B., et al. (2000). Functional properties of human ribonuclease fusion proteins expressed in Escherichia coli or transgenic mice. J. Int. Soc. Tumor Targ. 1, 70–81.
Newton D. L., Hansen H. J., Mikulski S. M., Goldenberg D. M., and Rybak S. M. (2001). Potent and specific antitumor effects of an anti-CD22 targeted cytotoxic ribonuclease: potential for the treatment of non-Hodgkin’s lymphoma. Blood 97, 528–535.
Horton R. M., Cai Z. L., Ho S. N., and Pease L. R. (1990). Gene splicing by overlap exension: tailor made genes using the polymerase chain reaction. BioTechniques 8, 528–535.
Studier F. W., Rosenberg A. H., Dunn J. J., and Dubendorff J. W. (1990). Use of T RNA polymerase to direct expression of cloned genes. Methods Enzymol. 185, 60–89.
Futami J., Tsushima Y., Tada H., Seno M., and Yamada H. (2000). Convenient and efficient in vitro folding of disulfide-containing globular protein from crude bacterial inclusion bodies. J. Biochem (Tokyo) 127, 435–441.
Seno M., DeSantis M., Kannan S., Bianco C., Tada H., Kim N., et al. (1998). Purification and characterization of a recombinant human cripto-1 protein. Growth Factors 15, 215–229.
Inoue M., Akimaru J., Nishikawa T., Seki N., and Yamada H. (1998). A new derivatizing agent, trimethylammoniopropyl methanethiosulphonate, is efficient for preparation of recombinant brain-derived neurotrophic factor from inclusion bodies. Biotechnol. Appl. Biochem. 28, 207–213.
Terzyan S. S., Peracaula R., de Llorens R., Tsushima Y., Yamada H., Seno M., et al. (1999). The three-dimensional structure of human RNase 4, unliganded and complexed with d(Up), reveals the basis for its uridine selectivity. J. Mol. Biol. 285, 205–214.
Mallorqui-Fernandez G., Pous J., Peracaula R., Aymami J., Maeda T., Tada H., et al. (2000). Three-dimensional crystal structure of human eosinophil cationic protein (RNase 3) at 1.75 A resolution. J. Mol. Biol. 300, 1297–1307.
Sambrook J., Fritsch E. F., and Maniatis T. (1989) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Harlow E. and Lane D. (1988) Antibodies: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Bond M. D. (1988). An in vitro binding assay for angiogenin using placental ribonuclease inhibitor. Anal. Biochem. 173, 166–173.
Gart J., Krewski D., Lee P., Tarone R., and Wahrendorf J. (1986) Statistical Methods in Cancer Research. International Agency for Research on Cancer, NY.
Tai M. S., Mudgett-Hunter M., Levinson D., Wu G.-M., Haber E., Oppermann H., and Huston J. S. (1990). A bifunctional fusion protein containing Fc-binding fragment B of staphylococcal protein A amino terminal to antidigoxin single-chain Fv. Biochemistry 29, 8024–8030.
Buchner J., Pastan I., and Brinkmann U. (1992). A method for increasing the yield of properly folded recombinant fusion proteins: Single-chain immunotoxins from renaturation of bacterial inclusion bodies. Anal. Biochem. 205, 263–270.
Crothers D. M. and Metzger H. (1972). The influence of polyvalency on the binding properties of antibodies. Immunochem. 9, 341–357.
Rosenberg H. F. and Dyer K. D. (1995). Eosinophil cationic protein and eosinophilderived neurotoxin. Evolution of novel function in a primate ribonuclease gene family. J. Biol. Chem. 270, 21,539–21,544.
Giovanella B. C., Stehlin J. S., Shepard R. C., and Williams L. J. (1979). Hyperthermic treatment of human tumors heterotransplanted in nude mice. Cancer Res. 39, 2236–2241.
Russo N., Nobile V., DiDonato A., Riordan J. F., and Valee B. L. (1996). The C-terminal region of human angiogenin has a dual role in enzymatic activity. Proc. Natl. Acad. Sci. USA 93, 3243–3247.
Mosimann S. C., Ardelt W., and James M. N. G. (1994). Refined 1.7 A X-ray crystallographic structure of P-30 protein, an amphibian ribonuclease with anti-tumor activity. J. Mol. Biol. 236, 1141–1153.
Boix E., Wu Y., Vasandani V. M., Saxena S. K., Ardelt W., Ladner J., and Youle R. J. (1996). Role of the N terminus in RNase A homologues: Differences in catalytic activity, ribonuclease inhibitor interaction and cytotoxicity. J. Mol. Biol. 257, 992–1007.
Newton D. L., Xue Y., Boque L., Wlodawer A., Kung H. F., and Rybak S. M. (1997). Expression and characterization of a cytotoxic human-frog chimeric ribonuclease: Potential for cancer therapy. Protein Eng. 10, 463–470.
Wehrli W., Knusel F., Schmid K., and Staehelin M. (1968). Interaction of rifamycin with bacterial RNA polymerase. Proc. Natl. Acad. Sci. USA 61, 667–673.
Goldenberg D. M., Horowitz J. A., Sharkey R. M., Hall T. C., Murthy S., Goldenberg H., et al. (1991). Targeting, dosimetry, and radioimmunotherapy of B-cell lymphomas with iodine-131-labeled LL2 monoclonal antibody. J. Clin. Oncol. 9, 548–564.
Ghetie M. A. J., 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 lymphoma grown in mice with severe combined immunodeficiency disease effect on tumor cells in extranodal sites. Cancer Res. 51, 5876–5880.
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.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.
About this protocol
Cite this protocol
Newton, D.L., Futami, J., Ruby, D., Rybak, S.M. (2003). Construction and Characterization of RNase-Based Targeted Therapeutics. In: Welschof, M., Krauss, J. (eds) Recombinant Antibodies for Cancer Therapy. Methods in Molecular Biology™, vol 207. Humana Press. https://doi.org/10.1385/1-59259-334-8:283
Download citation
DOI: https://doi.org/10.1385/1-59259-334-8:283
Publisher Name: Humana Press
Print ISBN: 978-0-89603-918-6
Online ISBN: 978-1-59259-334-7
eBook Packages: Springer Protocols