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. 1996 Sep;74(5):735–744. doi: 10.1038/bjc.1996.430

Generation of chimeric bispecific G250/anti-CD3 monoclonal antibody, a tool to combat renal cell carcinoma.

R M Luiten 1, L R Coney 1, G J Fleuren 1, S O Warnaar 1, S V Litvinov 1
PMCID: PMC2074712  PMID: 8795576

Abstract

The monoclonal antibody (MAb) G250 binds to a tumour-associated antigen, expressed in renal cell carcinoma (RCC), which has been demonstrated to be a suitable target for antibody-mediated immunotherapy. A bispecific antibody having both G250 and anti-CD3 specificity can cross-link G250 antigen-expressing RCC target cells with T cells and can mediate lysis of such targets. Therapy studies with murine antibodies are limited by immune responses to the antibodies injected (HAMA response), which can be decreased by using chimeric antibodies. We generated a chimeric bispecific G250/anti CD3 MAb by transfecting chimeric genes of heavy and light chains for both the G250 MAb and the anti-CD3 MAb into a myeloma cell line. Cytotoxicity assays revealed that the chimeric bispecific MAb was capable of mediating lysis of RCC cell lines by cloned human CD8+T cells or by IL-2-stimulated peripheral blood lymphocytes (PBLs). Lysis mediated by the MAb was specific for target cells that expressed the G250 antigen and was effective at concentrations as low as 0.01 microgram ml-1. The chimeric bispecific G250/anti-CD3 MAb produced may be an effective adjuvant to the currently used IL-2-based therapy of advanced renal cell arcinoma.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abrams J. S., Rayner A. A., Wiernik P. H., Parkinson D. R., Eisenberger M., Aronson F. R., Gucalp R., Atkins M. B., Hawkins M. J. High-dose recombinant interleukin-2 alone: a regimen with limited activity in the treatment of advanced renal cell carcinoma. J Natl Cancer Inst. 1990 Jul 18;82(14):1202–1206. doi: 10.1093/jnci/82.14.1202. [DOI] [PubMed] [Google Scholar]
  2. Berg J., Lötscher E., Steimer K. S., Capon D. J., Baenziger J., Jäck H. M., Wabl M. Bispecific antibodies that mediate killing of cells infected with human immunodeficiency virus of any strain. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4723–4727. doi: 10.1073/pnas.88.11.4723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolhuis R. L., Lamers C. H., Goey S. H., Eggermont A. M., Trimbos J. B., Stoter G., Lanzavecchia A., di Re E., Miotti S., Raspagliesi F. Adoptive immunotherapy of ovarian carcinoma with bs-MAb-targeted lymphocytes: a multicenter study. Int J Cancer Suppl. 1992;7:78–81. [PubMed] [Google Scholar]
  4. Brissinck J., Demanet C., Moser M., Leo O., Thielemans K. Treatment of mice bearing BCL1 lymphoma with bispecific antibodies. J Immunol. 1991 Dec 1;147(11):4019–4026. [PubMed] [Google Scholar]
  5. Bukowski R. M., Sharfman W., Murthy S., Rayman P., Tubbs R., Alexander J., Budd G. T., Sergi J. S., Bauer L., Gibson V. Clinical results and characterization of tumor-infiltrating lymphocytes with or without recombinant interleukin 2 in human metastatic renal cell carcinoma. Cancer Res. 1991 Aug 15;51(16):4199–4205. [PubMed] [Google Scholar]
  6. Courtenay-Luck N. S., Epenetos A. A., Moore R., Larche M., Pectasides D., Dhokia B., Ritter M. A. Development of primary and secondary immune responses to mouse monoclonal antibodies used in the diagnosis and therapy of malignant neoplasms. Cancer Res. 1986 Dec;46(12 Pt 1):6489–6493. [PubMed] [Google Scholar]
  7. De Lau W. B., Heije K., Neefjes J. J., Oosterwegel M., Rozemuller E., Bast B. J. Absence of preferential homologous H/L chain association in hybrid hybridomas. J Immunol. 1991 Feb 1;146(3):906–914. [PubMed] [Google Scholar]
  8. Ernstoff M. S., Gooding W., Nair S., Bahnson R. R., Miketic L. M., Banner B., Day R., Whiteside T., Titus-Ernstoff L., Kirkwood J. M. Immunological effects of treatment with sequential administration of recombinant interferon gamma and alpha in patients with metastatic renal cell carcinoma during a phase I trial. Cancer Res. 1992 Feb 15;52(4):851–856. [PubMed] [Google Scholar]
  9. Fanger M. W., Segal D. M., Wunderlich J. R. Going both ways: bispecific antibodies and targeted cellular cytotoxicity. FASEB J. 1990 Aug;4(11):2846–2849. doi: 10.1096/fasebj.4.11.2199282. [DOI] [PubMed] [Google Scholar]
  10. Feruglio C., Zambello R., Trentin L., Bulian P., Franceschi T., Cetto G. L., Semenzato G. Cytotoxic in vitro function in patients with metastatic renal cell carcinoma before and after alpha-2b-interferon therapy. Effects of activation with recombinant interleukin-2. Cancer. 1992 May 15;69(10):2525–2531. doi: 10.1002/1097-0142(19920515)69:10<2525::aid-cncr2820691023>3.0.co;2-d. [DOI] [PubMed] [Google Scholar]
  11. Fisher R. I., Coltman C. A., Jr, Doroshow J. H., Rayner A. A., Hawkins M. J., Mier J. W., Wiernik P., McMannis J. D., Weiss G. R., Margolin K. A. Metastatic renal cancer treated with interleukin-2 and lymphokine-activated killer cells. A phase II clinical trial. Ann Intern Med. 1988 Apr;108(4):518–523. doi: 10.7326/0003-4819-108-4-518. [DOI] [PubMed] [Google Scholar]
  12. Foon K. A., Walther P. J., Bernstein Z. P., Vaickus L., Rahman R., Watanabe H., Sweeney J., Park J., Vesper D., Russell D. Renal cell carcinoma treated with continuous-infusion interleukin-2 with ex vivo-activated killer cells. J Immunother (1991) 1992 Apr;11(3):184–190. doi: 10.1097/00002371-199204000-00005. [DOI] [PubMed] [Google Scholar]
  13. Garrido M. A., Valdayo M. J., Winkler D. F., Titus J. A., Hecht T. T., Perez P., Segal D. M., Wunderlich J. R. Targeting human T-lymphocytes with bispecific antibodies to react against human ovarian carcinoma cells growing in nu/nu mice. Cancer Res. 1990 Jul 15;50(14):4227–4232. [PubMed] [Google Scholar]
  14. Gaynor E. R., Weiss G. R., Margolin K. A., Aronson F. R., Sznol M., Demchak P., Grima K. M., Fisher R. I., Boldt D. H., Doroshow J. H. Phase I study of high-dose continuous-infusion recombinant interleukin-2 and autologous lymphokine-activated killer cells in patients with metastatic or unresectable malignant melanoma and renal cell carcinoma. J Natl Cancer Inst. 1990 Sep 5;82(17):1397–1402. doi: 10.1093/jnci/82.17.1397. [DOI] [PubMed] [Google Scholar]
  15. Hayat K., Rodgers S., Bruce L., Rees R. C., Chapman K., Reeder S., Dorreen M. S., Sheridan E., Sreenivasan T., Hancock B. W. Malignant melanoma and renal cell carcinoma: immunological and haematological effects of recombinant human interleukin-2. Eur J Cancer. 1991;27(8):1009–1014. doi: 10.1016/0277-5379(91)90270-n. [DOI] [PubMed] [Google Scholar]
  16. Jensen P. B., Birch C. Effects of halting a cervical screening programme. Lancet. 1990 Jun 23;335(8704):1523–1523. doi: 10.1016/0140-6736(90)93058-w. [DOI] [PubMed] [Google Scholar]
  17. Krigel R. L., Padavic-Shaller K. A., Rudolph A. R., Konrad M., Bradley E. C., Comis R. L. Renal cell carcinoma: treatment with recombinant interleukin-2 plus beta-interferon. J Clin Oncol. 1990 Mar;8(3):460–467. doi: 10.1200/JCO.1990.8.3.460. [DOI] [PubMed] [Google Scholar]
  18. Kroesen B. J., ter Haar A., Spakman H., Willemse P., Sleijfer D. T., de Vries E. G., Mulder N. H., Berendsen H. H., Limburg P. C., The T. H. Local antitumour treatment in carcinoma patients with bispecific-monoclonal-antibody-redirected T cells. Cancer Immunol Immunother. 1993 Nov;37(6):400–407. doi: 10.1007/BF01526797. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. LoBuglio A. F., Wheeler R. H., Trang J., Haynes A., Rogers K., Harvey E. B., Sun L., Ghrayeb J., Khazaeli M. B. Mouse/human chimeric monoclonal antibody in man: kinetics and immune response. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4220–4224. doi: 10.1073/pnas.86.11.4220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Luiten R. M., Fleuren G. J., Warnaar S. O., Litvinov S. V. Target-specific activation of mast cells by immunoglobulin E reactive with a renal cell carcinoma-associated antigen. Lab Invest. 1996 Feb;74(2):467–475. [PubMed] [Google Scholar]
  21. Marumo K., Muraki J., Ueno M., Tachibana M., Deguchi N., Baba S., Jitsukawa S., Hata M., Tazaki H. Immunologic study of human recombinant interleukin-2 (low-dose) in patients with advanced renal cell carcinoma. Urology. 1989 Mar;33(3):219–225. doi: 10.1016/0090-4295(89)90396-8. [DOI] [PubMed] [Google Scholar]
  22. Meredith R. F., LoBuglio A. F., Plott W. E., Orr R. A., Brezovich I. A., Russell C. D., Harvey E. B., Yester M. V., Wagner A. J., Spencer S. A. Pharmacokinetics, immune response, and biodistribution of iodine-131-labeled chimeric mouse/human IgG1,k 17-1A monoclonal antibody. J Nucl Med. 1991 Jun;32(6):1162–1168. [PubMed] [Google Scholar]
  23. Mezzanzanica D., Canevari S., Ménard S., Pupa S. M., Tagliabue E., Lanzavecchia A., Colnaghi M. I. Human ovarian carcinoma lysis by cytotoxic T cells targeted by bispecific monoclonal antibodies: analysis of the antibody components. Int J Cancer. 1988 Apr 15;41(4):609–615. doi: 10.1002/ijc.2910410422. [DOI] [PubMed] [Google Scholar]
  24. Muss H. B., Costanzi J. J., Leavitt R., Williams R. D., Kempf R. A., Pollard R., Ozer H., Zekan P. J., Grunberg S. M., Mitchell M. S. Recombinant alfa interferon in renal cell carcinoma: a randomized trial of two routes of administration. J Clin Oncol. 1987 Feb;5(2):286–291. doi: 10.1200/JCO.1987.5.2.286. [DOI] [PubMed] [Google Scholar]
  25. Nisticò P., Mortarini R., De Monte L. B., Mazzocchi A., Mariani M., Malavasi F., Parmiani G., Natali P. G., Anichini A. Cell retargeting by bispecific monoclonal antibodies. Evidence of bypass of intratumor susceptibility to cell lysis in human melanoma. J Clin Invest. 1992 Sep;90(3):1093–1099. doi: 10.1172/JCI115925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Oosterwijk E., Bander N. H., Divgi C. R., Welt S., Wakka J. C., Finn R. D., Carswell E. A., Larson S. M., Warnaar S. O., Fleuren G. J. Antibody localization in human renal cell carcinoma: a phase I study of monoclonal antibody G250. J Clin Oncol. 1993 Apr;11(4):738–750. doi: 10.1200/JCO.1993.11.4.738. [DOI] [PubMed] [Google Scholar]
  27. Oosterwijk E., Ruiter D. J., Hoedemaeker P. J., Pauwels E. K., Jonas U., Zwartendijk J., Warnaar S. O. Monoclonal antibody G 250 recognizes a determinant present in renal-cell carcinoma and absent from normal kidney. Int J Cancer. 1986 Oct 15;38(4):489–494. doi: 10.1002/ijc.2910380406. [DOI] [PubMed] [Google Scholar]
  28. Palmer P. A., Vinke J., Evers P., Pourreau C., Oskam R., Roest G., Vlems F., Becker L., Loriaux E., Franks C. R. Continuous infusion of recombinant interleukin-2 with or without autologous lymphokine activated killer cells for the treatment of advanced renal cell carcinoma. Eur J Cancer. 1992;28A(6-7):1038–1044. doi: 10.1016/0959-8049(92)90450-g. [DOI] [PubMed] [Google Scholar]
  29. Parkinson D. R., Fisher R. I., Rayner A. A., Paietta E., Margolin K. A., Weiss G. R., Mier J. W., Sznol M., Gaynor E. R., Bar M. H. Therapy of renal cell carcinoma with interleukin-2 and lymphokine-activated killer cells: phase II experience with a hybrid bolus and continuous infusion interleukin-2 regimen. J Clin Oncol. 1990 Oct;8(10):1630–1636. doi: 10.1200/JCO.1990.8.10.1630. [DOI] [PubMed] [Google Scholar]
  30. Qian J. H., Titus J. A., Andrew S. M., Mezzanzanica D., Garrido M. A., Wunderlich J. R., Segal D. M. Human peripheral blood lymphocytes targeted with bispecific antibodies release cytokines that are essential for inhibiting tumor growth. J Immunol. 1991 May 1;146(9):3250–3256. [PubMed] [Google Scholar]
  31. Riva P., Tison V., Arista A., Sturiale C., Franceschi G., Riva N., Casi M., Moscatelli G., Campori F., Spinelli A. Radioimmunotherapy of gastrointestinal cancer and glioblastomas. Int J Biol Markers. 1993 Jul-Sep;8(3):192–197. doi: 10.1177/172460089300800310. [DOI] [PubMed] [Google Scholar]
  32. Rosenberg S. A., Lotze M. T., Yang J. C., Aebersold P. M., Linehan W. M., Seipp C. A., White D. E. Experience with the use of high-dose interleukin-2 in the treatment of 652 cancer patients. Ann Surg. 1989 Oct;210(4):474–485. doi: 10.1097/00000658-198910000-00008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Saleh M. N., Khazaeli M. B., Grizzle W. E., Wheeler R. H., Lawson S., Liu T., Russel C., Meredith R., Schlom J., LoBuglio A. F. A phase I clinical trial of murine monoclonal antibody D612 in patients with metastatic gastrointestinal cancer. Cancer Res. 1993 Oct 1;53(19):4555–4562. [PubMed] [Google Scholar]
  34. Schroff R. W., Foon K. A., Beatty S. M., Oldham R. K., Morgan A. C., Jr Human anti-murine immunoglobulin responses in patients receiving monoclonal antibody therapy. Cancer Res. 1985 Feb;45(2):879–885. [PubMed] [Google Scholar]
  35. Shawler D. L., Bartholomew R. M., Smith L. M., Dillman R. O. Human immune response to multiple injections of murine monoclonal IgG. J Immunol. 1985 Aug;135(2):1530–1535. [PubMed] [Google Scholar]
  36. Tibben J. G., Boerman O. C., Claessens R. A., Corstens F. H., van Deuren M., de Mulder P. H., van der Meer J. W., Keijser K. G., Massuger L. F. Cytokine release in an ovarian carcinoma patient following intravenous administration of bispecific antibody OC/TR F(ab')2. J Natl Cancer Inst. 1993 Jun 16;85(12):1003–1004. doi: 10.1093/jnci/85.12.1003. [DOI] [PubMed] [Google Scholar]
  37. Warnaar S. O., De Paus V., Lardenoije R., Machielse B. N., De Graaf J., Bregonje M., Van Haarlem H. Purification of bispecific F(ab')2 from murine trinoma OC/TR with specificity for CD3 and ovarian cancer. Hybridoma. 1994 Dec;13(6):519–526. doi: 10.1089/hyb.1994.13.519. [DOI] [PubMed] [Google Scholar]
  38. Whitehead R. P., Ward D., Hemingway L., Hemstreet G. P., 3rd, Bradley E., Konrad M. Subcutaneous recombinant interleukin 2 in a dose escalating regimen in patients with metastatic renal cell adenocarcinoma. Cancer Res. 1990 Oct 15;50(20):6708–6715. [PubMed] [Google Scholar]
  39. van Dijk J., Oosterwijk E., van Kroonenburgh M. J., Jonas U., Fleuren G. J., Pauwels E. K., Warnaar S. O. Perfusion of tumor-bearing kidneys as a model for scintigraphic screening of monoclonal antibodies. J Nucl Med. 1988 Jun;29(6):1078–1082. [PubMed] [Google Scholar]
  40. van Dijk J., Uemura H., Beniers A. J., Peelen W. P., Zegveld S. T., Fleuren G. J., Warnaar S. O., Oosterwijk E. Therapeutic effects of monoclonal antibody G250, interferons and tumor necrosis factor, in mice with renal-cell carcinoma xenografts. Int J Cancer. 1994 Jan 15;56(2):262–268. doi: 10.1002/ijc.2910560220. [DOI] [PubMed] [Google Scholar]
  41. van Dijk J., Warnaar S. O., van Eendenburg J. D., Thienpont M., Braakman E., Boot J. H., Fleuren G. J., Bolhuis R. L. Induction of tumor-cell lysis by bi-specific monoclonal antibodies recognizing renal-cell carcinoma and CD3 antigen. Int J Cancer. 1989 Feb 15;43(2):344–349. doi: 10.1002/ijc.2910430230. [DOI] [PubMed] [Google Scholar]
  42. van Dijk J., Zegveld S. T., Fleuren G. J., Warnaar S. O. Localization of monoclonal antibody G250 and bispecific monoclonal antibody CD3/G250 in human renal-cell carcinoma xenografts: relative effects of size and affinity. Int J Cancer. 1991 Jul 9;48(5):738–743. doi: 10.1002/ijc.2910480518. [DOI] [PubMed] [Google Scholar]
  43. van Ravenswaay Claasen H. H., van de Griend R. J., Mezzanzanica D., Bolhuis R. L., Warnaar S. O., Fleuren G. J. Analysis of production, purification, and cytolytic potential of bi-specific antibodies reactive with ovarian-carcinoma-associated antigens and the T-cell antigen CD3. Int J Cancer. 1993 Aug 19;55(1):128–136. doi: 10.1002/ijc.2910550123. [DOI] [PubMed] [Google Scholar]

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