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Cancer Immunology, Immunotherapy

, Volume 33, Issue 4, pp 210–216 | Cite as

Bispecific-monoclonal-antibody-directed lysis of ovarian carcinoma cells by activated human T lymphocytes

  • Susanna A. Möller
  • Ralph A. Reisfeld
Original articles

Summary

Two different bispecific hybrid antibodies were established by fusing a hybridoma producing monoclonal antibody (mAb) against the pancarcinoma antigen KS1/4 with either of the two hybridomas OKT3 and 9.3, secreting antibodies reactive with the T cell determinants CD3 and CD28, respectively. The KS1/4 antibody reacts with a 40-kDa cell-surface glycoprotein antigen that is expressed on the surface of a variety of adenocarcinoma cells, including ovarian carcinoma. The ability of the bispecific antibodies 9.3⋅KS1/4 and OKT3⋅KS1/4 to direct peripheral blood mononuclear cells (PBMC) specifically against OVCAR-3 ovarian carcinoma target cells was measured in a 4-h51Cr-release assay. The bispecific antibodies were four to six times more potent in killing the OVCAR-3 target cells when compared to their parental antibodies either alone or in combination. A dose-dependent response was observed in the 10–10000 ng/ml range. The specificity of the targeting was demonstrated by the complete inhibition of cytotoxic activity following pre-incubation of tumor target cells with the parental mAb and by the lack of killing of KS1/4-negative target cell lines. An evaluation of the efficacy of PBMC from ovarian cancer patients as effector cells revealed that their specific cytotoxicity against OVCAR-3 cells was enhanced severalfold by bispecific antibodies as compared to parental antibodies. Furthermore, stimulation of PBMC with immobilized CD3 and interleukin-2 for 4 days resulted in an enhanced directed killing of human ovarian carcinoma cells by human T effector cells and the bispecific antibodies.

Key words

Monoclonal antibody T lymphocytes Ovarian carcinoma 

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References

  1. 1.
    Barr IG, Miescher S, von Vliedner V, Bechegger F, Barras C, Lanzavecchia A, Mach J-P, Carrel S (1989) In vivo localization of a bispecific antibody which targets human T lymphocytes to lyse human colon cancer cells. Int J Cancer 43: 501PubMedGoogle Scholar
  2. 2.
    Borrebaeck CAK, Möller SA (1986) In vitro immunization. Effect of growth and differentiation factors on antigen-specific B cell activation and production of monoclonal antibodies to autologous antigens and weak immunogens. J Immunol 136: 3710PubMedGoogle Scholar
  3. 3.
    Brodeur GM, Sekhon GS, Goldstein MN (1977) Chromosomal abberations in human neuroblastomas. Cancer 40: 2256PubMedGoogle Scholar
  4. 4.
    Clark M, Gilliland L, Waldmann H (1988) Hybrid antibodies for therapy. Prog Allergy 45: 31PubMedGoogle Scholar
  5. 5.
    van Dijk J, Warnaar SO, van Eedenburg JDH, Thienpont M, Braakman E, Boot JHA, Fleuren GJ, Bolhuis RLH (1989) Induction of tumor-cell lysis by bispecific monoclonal antibodies recognizing renal-cell carcinoma and CD3 antigen. Int J Cancer 43: 344PubMedGoogle Scholar
  6. 6.
    Hamilton TC, Young RC, Louie KG, Behrens BC, McKoy WM, Grotzinger KR, Ozols RF (1984) Characterization of a xenograft model of human ovarian carcinoma which produces ascites and intraabdominal carcinomatosis in mice. Cancer Res 44: 5286PubMedGoogle Scholar
  7. 7.
    Hansen JA, Martin PJ, Nowinski RC (1980) Monoclonal antibodies identifying a novel T cell antigen and Ia antigens of human lymphocytes. Immunogenetics 10: 247Google Scholar
  8. 8.
    Hansen JA, Martin PJ, Beatty PG, Clark EA, Ledbetter JA (1984) Human T lymphocyte cell surface molecules defined by the workshop monoclonal antibodies. In: Leucocyte typing. Springer, Berlin Heidelberg New York, p 195Google Scholar
  9. 9.
    Harper JR, Bumol TF, Reisfeld RA (1982) Serological and biochemical analyses of monoclonal antibodies to human melanoma-associated antigens. Hybridoma 1: 423PubMedGoogle Scholar
  10. 10.
    June CH, Ledbetter JA, Linsley PS, Thompson CB (1990) Role of CD28 receptor in T-cell activation. Immunol Today 11: 211PubMedGoogle Scholar
  11. 11.
    Jung G, Ledbetter JA, Müller-Eberhard HJ (1987) Induction of cytotoxicity in resting human T lymphocytes bound to tumor cells by antibody heteroconjugates. Proc Natl Sci USA 84: 4611Google Scholar
  12. 12.
    Kerr L, Huntoon C, Donjue J, Leibson PJ, Bander NH, Ghose T, Luner SJ, Vessella R, McKean DJ (1990) Heteroconjugate antibody-directed killing of autologous human renal carcinoma cells by in vitro-activated lymphocytes. J Immunol 144: 4060PubMedGoogle Scholar
  13. 13.
    Lanzavecchia A, Scheidegger D (1987) The use of hybrid hybridomas to target human cytotoxic T lymphocytes. Eur J Immunol 17: 105PubMedGoogle Scholar
  14. 14.
    Ledbetter JA, Martin PJ, Spooner CE, Wofsy D, Tsu TT, Beatty PG, Gladstone P (1985) Antibodies to Tp67 and Tp44 augment and sustain proliferative responses to activated T cells. J Immunol 135: 2331PubMedGoogle Scholar
  15. 15.
    Liu MA, Kranz DM, Kurnick JT, Boyle LA, Levy R, Eisen H (1985) Heteroantibody duplexes target cells for lysis by cytotoxic T lymphocytes. Proc Natl Acad Sci USA 82: 8648PubMedGoogle Scholar
  16. 16.
    Mezzanzanica D, Canevari S, Ménard S, Pupa SM, Tagliabue E, Lanzavecchia A, Colnaghi MI (1988) Human ovarian carcinoma lysis by cytotoxic T cells targeted by bispecific monoclonal antibodies: analysis of the antibody components. Int J Cancer 41: 609PubMedGoogle Scholar
  17. 17.
    Milstein C, Cuello AC (1983) Hybrid hybridomas and their use in immunohistochemistry. Nature 305: 537PubMedGoogle Scholar
  18. 18.
    Perez MS, Walker LE (1989) Isolation and characterization of a cDNA encoding the KS1/4 epithelial carcinoma marker. J Immunol 142: 3662PubMedGoogle Scholar
  19. 19.
    Perez P, Hoffman RW, Shaw S, Bluestone JA, Segal DM (1985) Specific targeting of cytotoxic T cells by anti-T3 linked to anti-target cell antibody. Nature 316: 354PubMedGoogle Scholar
  20. 20.
    Perez P, Titus JA, Lotze MT, Cuttitta F, Longo DL, Groves ES, Rabin H, Durba JP, Segal DM (1986) Specific lysis of human tumor cells by T cells coated with anti-T3 cross-linked to anti-tumor antibody. J Immunol 137: 2069PubMedGoogle Scholar
  21. 21.
    Pupa SM, Canevari S, Fontanelli R, Ménard S, Mezzanzanica D, Lanzavecchia A, Colnaghi MI (1988) Activation of mononuclear cells to be used for hybrid monoclonal antibody-induced lysis of human ovarian carcinoma cells. Int J Cancer 42: 455PubMedGoogle Scholar
  22. 22.
    Scott CF, Bolender S, McIntyre GD, Holldack J, Lambert JM, Venkatesh YP, Morimoto C, Ritz J, Schlossman SF (1989) Activation of human cytolytic cells through CD2/T11. Comparison of the requirements for the induction and direction of lysis of tumor targets by T cells and NK cells. J Immunol 142: 4105PubMedGoogle Scholar
  23. 23.
    Segal DM, Wunderlich JR (1988) Targeting of cytotoxic cells with heterocrosslinked antibodies. Cancer Invest 6: 83PubMedGoogle Scholar
  24. 24.
    Segal DM, Garrido MA, Perez P, Titus JA, Winkler DA, Ring DB, Kaubisch A, Wunderlich JR (1988) Targeted cytotoxic cells as a novel form of cancer immunotherapy. Mol Immunol 25: 1099PubMedGoogle Scholar
  25. 25.
    Staerz UD, Bevan MJ (1986) Hybrid hybridoma producing a bispecific monoclonal antibody that can focus effector T-cell activity. Proc Natl Acad Sci USA 83: 1453PubMedGoogle Scholar
  26. 26.
    Staerz UD, Kanagawa O, Bevan MJ (1985) Hybrid antibodies can target sites for attack by T cells. Nature 314: 628PubMedGoogle Scholar
  27. 27.
    Strnad J, Hamilton AE, Beavers LS, Gamboa GC, Apelgren LD, Taber LD, Sportsman JR, Bumol TF, Sharp JD, Gadski RA (1989) Molecular cloning and characterization of a human adenocarcinoma/epithelial cell surface antigen complementary DNA. Cancer Res 49: 314PubMedGoogle Scholar
  28. 28.
    Titus JA, Perez P, Kaubisch A, Garrido MA, Segal DM (1987) Human K/natural killer cells targeted with hetero-cross-linked antibodies specifically lyse tumor cells in vitro and prevent tumor growth in vivo. J Immunol 139: 3153PubMedGoogle Scholar
  29. 29.
    Varki NM, Reisfeld RA, Walker LE (1984) Antigens associated with a human lung cancer adenocarcinoma defined by monoclonal antibodies. Cancer Res 44: 681PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Susanna A. Möller
    • 1
  • Ralph A. Reisfeld
    • 1
  1. 1.Department of ImmunologyScripps Clinic and Research FoundationLa JollaUSA

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