Skip to main content
SpringerLink
Log in

Antigen forks: bispecific reagents that inhibit cell growth by binding selected pairs of tumor antigens

  • Original Articles
  • Bispecific Antibodies, Tumor-Associated Antigens, Cell Growth Inhibition
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Bispecific antibodies of a new category, termed “antigen forks”, were constructed by crosslinking antibodies that recognized pairs of distinct tumor cell surface antigens. At concentrations of 1–100 nM, several such forks inhibited the growth of human tumor cell lines bearing both relevant antigens. The same cells were not inhibited by unconjugated component antibodies, and the active conjugates did not inhibit the growth of human cell lines that expressed lower levels of relevant antigens. The three most active antigen forks all contained monoclonal antibody 454A12, which recognizes human transferrin receptor. This antibody was conjugated respectively to antibodies 113F1 (against a tumor-associated glycoprotein complex), 317G5 (against a 42-kDa tumor-associated glycoprotein), or 520C9 (against the c-erbB-2 protooncogene product). The 317G5-454A12 fork strongly inhibited the HT-29 and SW948 human colorectal cancer cell lines, while the 113F1-454A12 fork was also effective against SW948. By designing forks against antigens of incompatible function that are co-expressed at high levels on tumor cells but not on normal tissues, it may be possible to generate reagents that inhibit tumor growth with enhanced selectivity.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bacus SS, Stancovski I, Huberman E, Chin D, Hurwitz E, Mills GB, Ullrich A, Sela M, Yarden Y (1992) Tumor-inhibitory monoclonal antibodies to the HER2/neu receptor induce differentiation of human breast cancer cells. Cancer Res 52: 2580–2589

    PubMed  Google Scholar 

  2. Bergsagel PL, Victor-Kobrin C, Timblin CR, Trepel J, Kuehl WM (1992) A murine cDNA encodes a pan-epithelial glycoprotein that is also expressed on plasma cells. J Immunol 148: 590–596

    PubMed  Google Scholar 

  3. Bjorn MJ, Ring DB, Frankel AE (1985) Evaluation of monoclonal antibodies for the development of breast cancer immunotoxins. Cancer Res 45: 1214–1221

    PubMed  Google Scholar 

  4. Brennan M, Davison PF, Paulus H (1985) Preparation of bispecific antibodies by chemical recombination of monoclonal immunoglobulin G1 fragments. Science 229: 81–83

    PubMed  Google Scholar 

  5. Frankel AE, Ring DB, Tringale F, Hsieh-Ma ST (1985) Tissue distribution of breast cancer associated antigens defined by monoclonal antibodies. J Biol Response Mod 4: 273–286

    PubMed  Google Scholar 

  6. Gatter KC, Brown G, Trowbridge IS, Woolston R-E, Mason DY (1983) Transferrin receptors in human tissues: their distribution and possible clinical relevance. J Clin Pathol 36: 539–545

    PubMed  Google Scholar 

  7. Glennie MJ, McBride HM, Worth AT, Stevenson GT (1987) Preparation and performance of bispecific F(ab' gamma)2 antibody containing thioether-linked Fab gamma fragments. J Immunol 139: 2367–2375

    PubMed  Google Scholar 

  8. Hamada H, Tsuruo T (1986) Functional role for the 170-to 180-kDa glycoprotein specific to drug-resistant tumor cells as revealed by monoclonal antibodies. Proc Natl Acad Sci USA 83: 7785–7789

    PubMed  Google Scholar 

  9. Hilkens J, Ligtenberg MJL, Vos HL, Litvinov SV (1992) Cell membrane-associated mucins and their adhesion-modulating property. Trends Biochem Sci 17: 359–363

    PubMed  Google Scholar 

  10. Huang SS, Huang JS (1992) Purification and characterization of theneu/erbB2 ligand-growth factor from bovine kidney. J Biol Chem 267: 11508–11512

    PubMed  Google Scholar 

  11. Jefferies WA, Brandon MR, Hunt SV, Williams AF, Gatter KC, Mason DY (1984) Transferrin receptor on endothelium of brain capillaries. Nature 312: 162–163

    PubMed  Google Scholar 

  12. Karpovsky B, Titus JA, Stephany DA, Segal DM (1984) Production of target-specific effector cells using hetero-cross-linked aggregates containing anti-target cell and anti-Fc gamma receptor antibodies. J Exp Med 160: 1686–1701

    PubMed  Google Scholar 

  13. Kasprzyk PG, Song SU, Di FP, King CR (1992) Therapy of an animal model of human gastric cancer using a combination of antierbB-2 monoclonal antibodies. Cancer Res 52: 2771–2776

    PubMed  Google Scholar 

  14. Kemp JD, Smith KM, Kanner LJ, Gomez F, Thorson JA, Naumann PW (1990) Synergistic inhibition of lymphoid tumor growth in vitro by combined treatment with the iron chelator deferoxamine and an immunoglobulin G monoclonal antibody against the transferrin receptor. Blood 76: 991–995

    PubMed  Google Scholar 

  15. Kemp JD, Thorson JA, Stewart BC, Naumann PW (1992) Inhibition of hematopoietic tumor growth by combined treatment with deferoxamine and an IgG monoclonal antibody against the transferrin receptor: evidence for a threshold model of iron deprivation toxicity. Cancer Res 52: 4144–4148

    PubMed  Google Scholar 

  16. Lazarus AH, Baines MG (1985) A rapid and efficient microtechnique for the analysis of functional transferrin receptors on tumor cells. J Immunol Methods 79: 213–221

    PubMed  Google Scholar 

  17. Linsley PS, Ochs V, Laska S, Horn D, Ring DB, Frankel AE, Brown JP (1986) Elevated levels of a high molecular weight antigen detected by antibody W1 in sera from breast cancer patients. Cancer Res 46: 5444–5450

    PubMed  Google Scholar 

  18. Lupu R, Colomer R, Zugmaier G, Sarup J, Shepard M, Slamon D, Lippman ME (1990) Direct interaction of a ligand for the erbB2 oncogene product with the EGF receptor and p185erbB2. Science 249: 1552–1555

    PubMed  Google Scholar 

  19. Lupu R, Dickson RB, Lippman ME (1992) The role of erbB-2 and its ligands in growth control of malignant breast epithelium. J Steroid Biochem Mol Biol 43: 229–236

    PubMed  Google Scholar 

  20. Milstein C, Cuello AC (1983) Hybrid hybridomas, their use in immunohistochemistry. Nature 305: 537–540

    PubMed  Google Scholar 

  21. Milstein C, Cuello AC (1984) Hybrid hybridomas, the production of bi-specific monoclonal antibodies. Immunol Today 5: 299–304

    Google Scholar 

  22. Natali PG, Nicotra MR, Bigotti A, Venturo I, Slamon DJ, Fendly BM, Ulrich A (1990) Expression of the p185 encoded by HER2 oncogenen in normal and transformed human tissues. Int J Cancer 45: 457–461

    PubMed  Google Scholar 

  23. Peles E, Bacus SS, Koski RA, Lu HS, Wen D, Ogden SG, Levy RB, Yarden Y (1992) Isolation of theneu/HER-2 stimulatory ligand: a 44 kd glycoprotein that induces differentiation of mammary tumor cells. Cell 69: 205–216

    PubMed  Google Scholar 

  24. Reading CL (1984) Procedures for in vitro immunization and monoclonal antibody production. In: Baldwin TH, Allison JP (eds) Hybridomas and cellular immortality. Plenum Press, New York, p 235

    Google Scholar 

  25. Ring DB, Kassel JNA, Hsieh-Ma ST, Bjorn MJ, Tringale F, Eaton AM, Reid SA, Frankel AE, Nadji M (1989) Distribution and physical properties of BCA200, a 200000M r glycoprotein selectively associated with human breast cancer. Cancer Res 49: 3070–3080

    PubMed  Google Scholar 

  26. Ring DB, Clark R, Saxena A (1991) Identity of BCA200 and c-erbB-2 indicated by reactivity of monoclonal antibodies with recombinant c-erbB2. Mol Immunol 28: 915–917

    PubMed  Google Scholar 

  27. Shepard HM, Lewis GD, Sarup JC, Fendley BM, Maneval D, Mordenti J, Figari I, Kotts CE, Palladino MJ, Ullrich A, et al (1991) Monoclonal antibody therapy of human cancer: taking the HER2 protooncogene to the clinic. J Clin Immunol 11: 117–127

    PubMed  Google Scholar 

  28. Stancovski I, Hurwitz E, Leitner O, Ullrich A, Yarden Y, Sela M (1991) Mechanistic aspects of the opposing effects of monoclonal antibodies to theerbB2 receptor on tumor growth. Proc Natl Acad Sci USA 88: 8691–8695

    PubMed  Google Scholar 

  29. Stancovski I, Peles E, Ben LR, Lemprecht R, Kelman Z, Goldman MR, Hurwitz E, Bacus S, Sela M, Yarden Y (1992) Signal transduction by theneu/erbB-2 receptor: a potential target for antitumor therapy. J Steroid Biochem Mol Biol 43: 95–103

    PubMed  Google Scholar 

  30. Tagliabue E, Centis F, Campiglio M, Mastroianni A, Martignone S, Pellegrini R, Casalini P, Lanzi C, Menard S, Colnaghi MI (1991) Selection of monoclonal antibodies which induce intermalization and phosphorylation of p185HER2 and growth inhibition of cells with HER2/neu gene amplification. Intl J Cancer 47: 933–937

    Google Scholar 

  31. Thorstensen K, Romslo I (1990) The role of transferrin in the mechanism of cellular iron uptake. Biochem J 271: 1–10

    PubMed  Google Scholar 

  32. Trauth BC, Klas C, Peters AMJ, Matzku S, Moller P, Falk W, Debatin K-M, Krammer PH (1989) Monoclonal antibody-mediated tumor regression by induction of apoptosis. Science 245: 301–305

    PubMed  Google Scholar 

  33. Trowbridge IS (1989) Transferrin receptor as a potential therapeutic target. Prog Allergy 45: 121–146

    Google Scholar 

  34. Trowbridge IS (1991) Endocytosis and signals for internalization. Curr Opin Cell Biol 3: 634–641

    PubMed  Google Scholar 

  35. Trowbridge IS, Lopez F (1982) Monoclonal antibody to transterrin receptor blocks transferrin binding and inhibits human tumor cell growth in vitro. Proc Natl Acad Sci USA 79: 1175

    PubMed  Google Scholar 

  36. Trowbridge IS, Lesley J, Schulte R (1982) Murine cell surface transferrin receptor: studies with an anti-receptor monoclonal antibody. J Cell Physiol 112: 403–416

    PubMed  Google Scholar 

  37. White S, Taetle R, Seligman PA, Rutherford M, Trowbridge IS (1990) Combinations of anti-transferrin receptor monoclonal antibodies inhibit human tumor cell growth in vitro and in vivo: evidence for synergistic antiproliferative effects. Cancer Res 50: 6296–6301

    Google Scholar 

  38. Yarden Y, Peles E (1991) Biochemical analysis of the ligand for theneu oncogenic receptor. Biochemistry 30: 3543–3550

    PubMed  Google Scholar 

  39. Yonehara S, Ishii A, Yonehara M (1989) A cell-killing monoclonal antibody (anti-Fas) to a cell surface antigen co-downregulated with the receptor of tumor necrosis factor. J Exp Med 169: 1747–1756

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Immunotherapeutics, Chiron Corporation, 4560 Horton, 94608, Emeryville, CA, USA

    David B. Ring, Sylvia T. Hsieh-Ma, Tim Shi & John Reeder

Authors
  1. David B. Ring
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sylvia T. Hsieh-Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tim Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. John Reeder
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ring, D.B., Hsieh-Ma, S.T., Shi, T. et al. Antigen forks: bispecific reagents that inhibit cell growth by binding selected pairs of tumor antigens. Cancer Immunol Immunother 39, 41–48 (1994). https://doi.org/10.1007/BF01517179

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01517179

Key words

Search

Navigation