Skip to main content
SpringerLink
Log in

Diphtheria Toxin Fusion Proteins

  • Chapter
Clinical Applications of Immunotoxins

Part of the book series: Current Topics in Microbiology and Immunology ((CT MICROBIOLOGY,volume 234))

Abstract

Two different approaches have been undertaken to develop targeted biomolecules for therapeutics. The first was the construction of immunotoxins consisting of monoclonal antibodies chemically linked through a disulfide bond to a plant or bacterial toxin or radionuclide. Instability of the chemical conjugation of some of the earlier immunotoxins led to the concept of using protein engineering and recombinant DNA to assemble fusion genes combining the sequences for the enzymatically active and translocation domains of a toxin with those of a specific targeting ligand. From the outset, the prospect of using recombinant DNA methods to assemble the structural genes encoding bacterial toxin growth factor fusion toxins, or fusion proteins, offered significant advantages over chemical conjugation in the assembly of chimeric proteins. Most importantly, the fusion junction, or point at which the substitute receptor binding domain was linked to the toxin fragment, could be precisely determined. Expression of the fusion gene in recombinant Escherichia coli would then result in the synthesis of a single homogeneous gene product rather than the mixture of isomeric forms which result from the chemical conjugation process used in the generation of immunotoxins, thereby leading to a theoretically more uniform agent for clinical studies.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Aullo P, Alcani J, Popoff MR, Klatzman DR, Murphy JR, Boguet P (1992) In vitro effects of a recombinant diphtheria-human CD4 fusion toxin on acute and chronically HIV-I infected cells. EMBO J 12: 921–931

    Google Scholar 

  • Bacha P, Waters C, Williams J, Murphy JR, Strom TB (1988) Interleukin-2 targeted cytotoxicity: selective action of a diphtheria toxin-related interleukin-2 fusion toxin. J Exp Med 167: 612–622

    Article  PubMed  CAS  Google Scholar 

  • Barnett D, Wilson G, Lawrence A, Buckley G (1988) The interleukin-2 receptor and its expression in the acute leukemias and lymphoproliferative disorders. Dis Markers 6: 133–139

    PubMed  CAS  Google Scholar 

  • Battaglia R, Scambia G, Benedetti P, Baiocchi G, Perrone L, Iaocabellli S, Mancuso S (1989) Epidermal growth factor receptor in gynecological malignancies. Gynecol Obstet Invest 27: 42

    Article  PubMed  CAS  Google Scholar 

  • Bennett MJ, Choe S, Eisenberg D (1994) Domain swapping: entangling alliances between proteins. Proc Natl Acad Sci USA 91: 3127–3131

    Article  PubMed  CAS  Google Scholar 

  • Berchuck A, Rodriguez G, Kamel A, Dodge R, Soper J, Clarke-Pearson D, Bast R (1991) Epidermal growth factor receptor expression in normal ovarian epithelium and ovarian cancer. Am J Obstet Gynecol 164: 669

    PubMed  CAS  Google Scholar 

  • Cain CC, Sipe DM, Murphy RF (1989) Regulation of endocytic pH by the Na+K+-ATPase in living cells. Proc Natl Acad Sci USA 86: 544–548

    Article  PubMed  CAS  Google Scholar 

  • Carpenter G, Chen S (1990) Epidermal growth factor. J Biol Chem 265: 7709

    PubMed  CAS  Google Scholar 

  • Choe S, Bennett M, Fujii G, Curmi P, Kantardjieff K, Collier R, Eisenberg D (1992) The crystal structure of diphtheria toxin. Nature 357: 216–222

    Article  PubMed  CAS  Google Scholar 

  • Craig F, Banks P (1992) Detection of the alpha and beta components of the interleukin-2 receptor using immunologic techniques. Mod Pathol 5: 118a

    Google Scholar 

  • Donovan JJ, Simon MI, Draper RK, Montal M (1981) Diphtheria toxin forms transmembrane channels in planar lipid bilayers. Proc Natl Acad Sci USA 78: 172–176

    Article  PubMed  CAS  Google Scholar 

  • Eisbruch A, Blick M, Lee J, Sacks P, Gutterman J (1987) Analysis of the epidermal growth factor receptor in fresh human head and neck tumors. Cancer Res 47: 3603

    PubMed  CAS  Google Scholar 

  • Foss F, Borkowski T, Gilliom M, Stetler-Stevenson M, Jaffe E, Tomkins A, Bastian A, Nylen P, Woodworth T, Udey M, Sausville E (1994) Chimeric fusion protein toxin DAB(486)IL2 in refractory mycosis fungoides and the sezary syndrome: correlation of activity and IL2 receptor expression in a phase II study. Blood 84 (6): 1765–1774

    PubMed  CAS  Google Scholar 

  • Fuchs R, Schmid S, Mellman I (1989) A possible role for Na+, K+-ATPase in regulating ATP-dependent endosome acidification. Proc Natl Acad Sci USA 86: 539–543

    Article  PubMed  CAS  Google Scholar 

  • Gottlieb AB, Lifshitz B, Fu SM, Staiano-Coico L, Wang CY, Carter DM (1986) Expression of HLA-DR molecules by keratinocytes and presence of Langerhans cells in the dermal infiltrate of active psoriatic plaques. J Exp Med 164: 1013–1028

    Article  PubMed  CAS  Google Scholar 

  • Gottlieb SL, Gilleaudeau P, Johnson R, Estes L, Woodworth TG, Gottlieb AB, Krueger JG (1995) Response of psoriasis to a lymphocyte-selective toxin (DAB389IL-2) suggests a primary immune, but not keratinocyte, pathogenic basis. Nature Med 1: 442–447

    Article  PubMed  CAS  Google Scholar 

  • Greenfield L, Johnson VG, Youle RJ (1987) Mutations in diphtheria toxin separate binding from entry and amplify immunotoxin selectivity. Science 238: 536–539

    Article  PubMed  CAS  Google Scholar 

  • Gupta AK, Ellis CN, Nickoloff BJ, Goldfarb MT, Ho VC, Rocher LL, Griffiths CEM, Cooper KD, Voorhees JJ (1990) Oral cyclosporine in the treatment of inflammatory and noninflammatory dermatoses. A clinical and immunopathologic analysis. Arch Dermatol 126: 339–350

    Article  PubMed  CAS  Google Scholar 

  • Hayakawa S, Uchida T, Mekada E, Moynihan MR, Okada Y (1983) Monoclonal antibody against diphtheria toxin: effect on toxin binding and entry into cells. J Biol Chem 258: 4311–4317

    PubMed  CAS  Google Scholar 

  • Hesketh P, Caguioa P, Koh H, Dewey H, Facada A, McCaffrey R, Parker K, Nylen P, Woodworth T (1993) Clinical activity of a cytotoxic fusion protein in the treatment of cutaneous T cell lymphoma. J Clin Oncol 11: 1628–1690

    Google Scholar 

  • Hoch DH, Romero-Mira M, Ehrich BE, Finkelstein A, DasGupta BR, Simpson LL (1985) Channels formed by botulinum, tetanus, and diphtheria toxins in planar lipid bilayers: relevance to translocation of proteins across membranes. Proc Natl Acad Sci USA 82: 336–343

    Article  Google Scholar 

  • Jean L-F, Murphy JR (1992) Diphtheria toxin receptor binding domain substitution with interleukin-6: genetic construction and interleukin-6 receptor specific action of a diphtheria toxin-related interleukin-6 fusion protein. Protein Eng 4: 989–994

    Article  Google Scholar 

  • Kagan BL, Finkelstein A, Colombini M (1981) Diphtheria toxin fragment forms large pores in phospholipid bilayer membranes. Proc Natl Acad Sci USA 78: 4950–4954

    Article  PubMed  CAS  Google Scholar 

  • Kiyokawa T, Williams DP, Snider CE, Strom TB, Murphy JR (1991) Protein engineering of diphtheria toxin-related interleukin-2 fusion toxins to increase biologic potency for high affinity interleukin-2 receptor bearing target cells. Protein Eng 4: 463–468

    Article  PubMed  CAS  Google Scholar 

  • Kraulis PJ (1991) MOLESCRIPT: a program to produce both detailed and schematic plots of protein structures. J Appl Crystallogr 24: 946–950

    Article  Google Scholar 

  • Kung E, Meissner K, Loning T (1988) Cutaneous T cell lymphoma: immunocytochemical study on activation/proliferation and differentiation associated antigens in lymph nodes, skin, and peripheral blood. Virchows Arch [A] Pathol Anat 413: 539–549

    Article  CAS  Google Scholar 

  • Lakkis F, Steele A, Pacheco-Silva A, Kelley VE, Strom TB, Murphy JR (1991) Interleukin-2 receptor targeted cytotoxicity: genetic construction and properties of diphtheria toxin-related interleukin-4 fusion toxins. Eur J Immunol 21: 2253–2258

    Article  PubMed  CAS  Google Scholar 

  • LeMaistre CF, Meneghetti C, Rosenblum M, Reuben J, Parker K, Shaw J, Woodworth T, Parkinson D (1992) Phase I trial of an interleukin-2 receptor (IL-2R) fusion toxin (DAB486 IL-2) in hematologic malignancies expressing the IL-2 receptor. Blood 79: 2547–2554

    PubMed  CAS  Google Scholar 

  • LeMaistre CF, Craig FE, Meneghetti C, McMullin B, Parker K, Reuben J, Boldt DH, Rosenblum M, Woodworth T (1993) Phase I trail of a 90-minute infusion of the fusion toxin DAB4F6 IL-2 in hematologic cancers. Cancer Res 53: 3930–3934

    PubMed  CAS  Google Scholar 

  • Lemoine N, Hughes C, Barton C, Poulson R, Jeffery R, Kloppel G, Ha11P, Gullick W (1992) The epidermal growth factor receptor in human pancreatic cancer. J Pathol 166: 7

    CAS  Google Scholar 

  • Middlebrook JL, Dorland RB, Leppla SH (1978) Association of diphtheria toxin with Vero cells: demonstration of a receptor. J Biol Chem 253: 7325–7330

    PubMed  CAS  Google Scholar 

  • Morris G, Dodd J (1990) Epidermal growth factor receptor mRNA levels in human prostatic tumors and cell lines. J Urol 143: 1272

    PubMed  CAS  Google Scholar 

  • Morris RE, Gerstein AS, Bonventre PF, Saelinger CB (1985) Receptor-mediated entry of diphtheria toxin into monkey kidney (Vero) cells: electron microscopic evaluation. Infect Immun 50: 721–727

    PubMed  CAS  Google Scholar 

  • Moya M, Dautry-Versat A, Goud B, Louvard D, Boquet P (1985) Inhibition of coated pit formation in Hep2 cells blocks the cytotoxicity of diphtheria toxin but not that of ricin toxin. J Cell Biol 101: 548–559

    Article  PubMed  CAS  Google Scholar 

  • Murphy JR, Pappenheimer AM Jr, Tayart de Borms S (1974) Synthesis of diphtheria tox gene products in Escherichia coli extracts. Proc Natl Acad Sci USA 71: 11–15

    Article  CAS  Google Scholar 

  • Murphy JR, Bishai W, Borowski M, Miyanohara A, Boyd J. Nagle S (1986) Genetic construction, expression, and melanoma-selective cytotoxicity of a diphtheria toxin a-melanocyte stimulating hormone fusion toxin. Proc Natl Acad Sci USA 83: 8258–8262

    Article  PubMed  CAS  Google Scholar 

  • Myers DA, Villemez CL (1988) Specific chemical cleavage of diphtheria toxin with hydroxyamine: purification and characterization of the modified proteins. J Biol Chem 263: 17122–17127

    PubMed  CAS  Google Scholar 

  • Neal D, Mellon K (1992) Epidermal growth factor receptor and bladder cancer. Urol Int 48: 365

    Article  PubMed  CAS  Google Scholar 

  • Nicholson S, Halcrow P, Sainsbury J, Angus B, Chambers P, Farndon J, Harris A (1988) Epidermal growth factor receptor status associated with failure of primary endocrine therapy in elderly postmenopausal patients with breast cancer. Br J Cancer 58: 810

    Article  PubMed  CAS  Google Scholar 

  • Nickoloff BJ, Griffiths CEM (1990) Lymphocyte trafficking in psoriasis: a new perspective emphasizing the dermal dendrocyte with active dermal recruitment mediated via endothelial cells followed by intra-epidermal T cell activation. J Invest Dermatol 95: 35S - 37S

    Article  Google Scholar 

  • Papini E, Sandond D, Rappuoli R, Montecucco C (1988) On the membrane translocation of diphtheria toxin: at low pH the toxin induces ion channels in cells. EMBO J 7: 3353–3359

    PubMed  CAS  Google Scholar 

  • Rolf JM, Eidels L (1993) Structure-function analyses of diphtheria toxin by use of monoclonal antibodies. Infect Immun 61: 944–1003

    Google Scholar 

  • Rosolen A, Nakanishi M, Poplack D, Cole D, Quinines R, Reaman G, Cotelingam J, Trepel J, Sausville E, Marti G, Neckers L, Colamonici O (1989) Expression of interleukin-2 receptor beta subunit in hematopoietic malignancies. Blood 73: 1968–1972

    PubMed  CAS  Google Scholar 

  • Sandvig K, Olsnes S (1988) Diphtheria toxin-induced channels in Vero cells selective for monovalent cations. J Biol Chem 263: 12352–12359

    PubMed  CAS  Google Scholar 

  • Schwartz G, Tepler I, Charette J, Kadin L, Parker K, Woodworth T, Schnipper L (1992) Complete response of a Hodgkin’s lymphoma in a phase I trail of DAB486 IL-2. Blood 79: 175a

    Google Scholar 

  • Shaw JP, Akiyoshi DE, Arrigo DA, Rhoad AE, Sullivan B, Thomas J, Genbauffe FS, Bacha P, Nichols JC (1991) Cytotoxic properties of DAB486 EGF and DAB389 EGF, epidermal growth factor ( EGF) receptor-targeted fusion toxins. J Biol Chem 266: 13449–13455

    Google Scholar 

  • Sheibani K, Winberg C, van de Velde S, Blayney D, Rappaport H (1987) Distribution of lymphocytes with interleukin-2 receptors (TAC antigens) in reactive lymphoproliferative processes, Hodgkins disease, and non-Hodgkins lymphomas. An immunohistochemical study of 300 cases. Am J Pathol 127: 27–37

    CAS  Google Scholar 

  • Smith K, Fennelly J, Neal D, Hall R, Harris A (1989) Characterization and quantitation of epidermal growth factor receptors in invasive and superficial bladder tumors. Cancer Res 49: 5810

    PubMed  CAS  Google Scholar 

  • Steele R, Kelly P, Ellul B, Ermin O (1990) Epidermal growth factor receptor expression in colorectal cancer. Br J Surg 77: 1352

    Article  PubMed  CAS  Google Scholar 

  • Strauchen J, Breakstone B (1987) IL-2 receptor expression in human lymphoid lesions. Am J Pathol 126: 506–512

    PubMed  CAS  Google Scholar 

  • Takeshita T, Asao H, Ohtani K, Ishii N, Kumaki S, Tanaka N, Munakata H, Nakamura M, Sugamura K (1992) Cloning of the chain of the human IL-2 receptor. Science 257: 379–382

    Article  PubMed  CAS  Google Scholar 

  • Theodoulou M, Baselga J, Scher H, Dantis L, Trainor K, Mendelsohn J, Bacha P, Brandt-Sarif T, Osborne K (1995) Phase I dose escalation study of the safety, tolerability, and pharmacokinetics of DAB389EGF in patients with solid malignancies expressing EGF receptors. Proc ASCO 14: 480

    Google Scholar 

  • Uchida T, Gill DM, Pappenheimer AM Jr (1971) Mutation in the structural gene for diphtheria toxin carried by temperate phage ß. Nature 233: 8–11

    Article  CAS  Google Scholar 

  • Uchida T, Pappenheimer AM Jr, Greaney R (1973) Diphtheria toxin and related proteins: isolation and properties of mutant proteins serologically related to diphtheria toxin. J Biol Chem 248: 3838–3844

    PubMed  CAS  Google Scholar 

  • Uchiyama T, Hori T, Tsudo M, Wano Y, Umadome H, Tamori S, Yodoi J, Maeda M, Sawami H, Uchino H (1985) Interleukin-2 receptor ( Tac antigen) expressed on adult T cell leukemia cells. Clin Invest 76: 446

    Article  CAS  Google Scholar 

  • Veale D, Ker N, Gibson G, Harris A (1989) Characterization of epidermal growth factor receptor in primary human non-small cell lung cancer. Cancer Res 49: 1313

    PubMed  CAS  Google Scholar 

  • Vyse TJ, Todd JA (1996) Genetic analysis of autoimmune disease. Cell 85: 311–318

    Article  PubMed  CAS  Google Scholar 

  • Waldmann TA (1986) The structure, function, and expression of interleukin-2 receptors on normal and malignant T cells. Science 232: 727–732

    Article  PubMed  CAS  Google Scholar 

  • Waldmann TA (1990) The multichain interleukin-2 receptor. A target for immunotherapy in lymphoma, autoimmune disorders, and organ allografts. J Am Med Assoc 263: 272–274

    Article  CAS  Google Scholar 

  • Walz G, Zanker B, Brand K, Swanlund D, Genbauffe F, Zeldis J, Murphy J, Strom T (1989) Sequential effects of interleukin-2/diphtheria toxin fusion protein on T cell activation. Proc Natl Acad Sci USA 86: 9485–9488

    Article  PubMed  CAS  Google Scholar 

  • Waters CA, Schimke P, Snider CE, Itoh K, Smith KA, Nichols JC, Strom TB, Murphy JR (1990) Interleukin-2 receptor targeted cytotoxicity: receptor binding requirements for entry of IL-2-toxin into cells. Eur J Immunol 20: 785–791

    Article  PubMed  CAS  Google Scholar 

  • Weidmann E, Sacchi M, Plaisance S, Heo D, Yasumura S, Lin W, Johnson J, Herberman R, Assarone B, Whiteside T (1992) Receptors for interleukin-2 on human squamous cell carcinoma cell lines and tumors in-situ. Cancer Res 52: 5963–5970

    PubMed  CAS  Google Scholar 

  • Weinstein GD, Krueger JG (1993) An overview of psoriasis. In: Weinstein GD, Gottlieb AB (eds) Therapy of moderate-to-severe psoriasis. National Psoriasis Foundation, Portland, OR, pp 1–22

    Google Scholar 

  • Wen Z, Tao X, Lakkis F, Kiyokawa T, Murphy JR (1991) Expression, purification, and a-melanocyte stimulating hormone receptor-specific toxicity of DAB-a-MSH fusion toxins. J Biol Chem 266: 12289–12293

    PubMed  CAS  Google Scholar 

  • Williams D, Parker K, Bishai W, Borowski M, Genbauffe F, Strom TB, Murphy JR (1987) Diphtheria-toxin receptor binding domain substitution with interleukin-2: genetic construction and properties of a diphtheria toxin-related interleukin-2 fusion protein. Protein Engn 1: 493–498

    Article  CAS  Google Scholar 

  • Williams D, Snider CE, Strom TB, Murphy JR (1990) Structure function analysis of IL-2 toxin (DAB486 IL-2): fragment B sequences required for the delivery of fragment A to the cytosol of target cells. J Biol Chem 265: 11885–11889

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Boston University Medical Center, 88 East Newton Street, Evans Bldg, Room 556, Boston, MA, 02118, USA

    F. M. Foss & J. R. Murphy

  2. Department of Medicine, University of Alabama School of Medicine, Birmingham, AL, USA

    M. N. Saleh

  3. Laboratory of Investigative Dermatology, Rockefeller University, New York, NY, USA

    J. G. Krueger

  4. Seragen, Inc., Hopkinton, MA, USA

    J. C. Nichols

Authors
  1. F. M. Foss
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. N. Saleh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. G. Krueger
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. C. Nichols
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J. R. Murphy
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Hollings Cancer Center, Medical University of South Carolina, 171 Ashley Avenue, 29425-2850, Charleston, SC, USA

    Arthur E. Frankel M.D. (Associate Professor of Medicine) (Associate Professor of Medicine)

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Springer-Verlag Berlin Heidelberg

About this chapter

Cite this chapter

Foss, F.M., Saleh, M.N., Krueger, J.G., Nichols, J.C., Murphy, J.R. (1998). Diphtheria Toxin Fusion Proteins. In: Frankel, A.E. (eds) Clinical Applications of Immunotoxins. Current Topics in Microbiology and Immunology, vol 234. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-72153-3_5

Download citation

  • DOI: https://doi.org/10.1007/978-3-642-72153-3_5

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-72155-7

  • Online ISBN: 978-3-642-72153-3

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation