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. 2001 Jul;85(1):129–136. doi: 10.1054/bjoc.2001.1891

Therapy of human non-small-cell lung carcinoma using antibody targeting of a modified superantigen

G Forsberg 1, L Ohlsson 1, T Brodin 1, P Björk 1, P A Lando 1, D Shaw 2, P L Stern 2, M Dohlsten 3
PMCID: PMC2363924  PMID: 11437414

Abstract

Superantigens activate T-cells by linking the T-cell receptor to MHC class II on antigen-presenting cells, and novel reactivity can be introduced by fusing the superantigen to a targeting molecule. Thus, an antibody-targeted superantigen, which activates T cells to destroy tumour cells, might be used as cancer therapy. A suitable target is the 5T4 oncofetal antigen, which is expressed on many carcinomas. We constructed a fusion protein from a Fab of a monoclonal antibody recognizing the 5T4 antigen, and an engineered superantigen. The recombinant product 5T4FabV13-SEAD227A bound the 5T4 antigen expressed on the human non-small-cell lung cancer cell line Calu-1 with a Kd of 1.2 nM while the substitution of Asp227 to Ala in the superantigen moiety reduced binding activity to MHC class II. 5T4FabV13-SEAD227A tumour reactivity was demonstrated in 7/7 NSCLC samples by immunohistochemistry, while normal tissue reactivity was low to moderate. 5T4FabV13-SEAD227A induced significant T-cell-dependent in vitro killing of sensitive 5T4 bearing Calu-1 cells, with maximum lysis at 10−10M, while the capacity to lyse MHC class II expressing cells was approximately 1000 times less effective. Immunotherapy of 5T4FabV13-SEAD227A against human NSCLC was investigated in SCID mice reconstituted with human peripheral blood mononuclear cells. Mice carrying intreperitoneally growing Calu-1 cells showed significant reduction in tumour mass and number after intravenous therapy with 5T4FabV13-SEAD227A. Thus, 5T4FabV13-SEAD227A has highly attractive properties for therapy of human NSCLC. © 2001 Cancer Research Campaign http://www.bjcancer.com

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Selected References

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  1. Abrahmsén L., Dohlsten M., Segrén S., Björk P., Jonsson E., Kalland T. Characterization of two distinct MHC class II binding sites in the superantigen staphylococcal enterotoxin A. EMBO J. 1995 Jul 3;14(13):2978–2986. doi: 10.1002/j.1460-2075.1995.tb07300.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bontkes H. J., de Gruijl T. D., Bijl A., Verheijen R. H., Meijer C. J., Scheper R. J., Stern P. L., Burns J. E., Maitland N. J., Walboomers J. M. Human papillomavirus type 16 E2-specific T-helper lymphocyte responses in patients with cervical intraepithelial neoplasia. J Gen Virol. 1999 Sep;80(Pt 9):2453–2459. doi: 10.1099/0022-1317-80-9-2453. [DOI] [PubMed] [Google Scholar]
  3. Bontkes H. J., van Duin M., de Gruijl T. D., Duggan-Keen M. F., Walboomers J. M., Stukart M. J., Verheijen R. H., Helmerhorst T. J., Meijer C. J., Scheper R. J. HPV 16 infection and progression of cervical intra-epithelial neoplasia: analysis of HLA polymorphism and HPV 16 E6 sequence variants. Int J Cancer. 1998 Oct 5;78(2):166–171. doi: 10.1002/(sici)1097-0215(19981005)78:2<166::aid-ijc8>3.0.co;2-x. [DOI] [PubMed] [Google Scholar]
  4. Dall'Acqua W., Carter P. Antibody engineering. Curr Opin Struct Biol. 1998 Aug;8(4):443–450. doi: 10.1016/s0959-440x(98)80121-8. [DOI] [PubMed] [Google Scholar]
  5. Dohlsten M., Abrahmsén L., Björk P., Lando P. A., Hedlund G., Forsberg G., Brodin T., Gascoigne N. R., Förberg C., Lind P. Monoclonal antibody-superantigen fusion proteins: tumor-specific agents for T-cell-based tumor therapy. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8945–8949. doi: 10.1073/pnas.91.19.8945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dohlsten M., Björklund M., Sundstedt A., Hedlund G., Samson D., Kalland T. Immunopharmacology of the superantigen staphylococcal enterotoxin A in T-cell receptor V beta 3 transgenic mice. Immunology. 1993 Aug;79(4):520–527. [PMC free article] [PubMed] [Google Scholar]
  7. Dohlsten M., Hansson J., Ohlsson L., Litton M., Kalland T. Antibody-targeted superantigens are potent inducers of tumor-infiltrating T lymphocytes in vivo. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9791–9795. doi: 10.1073/pnas.92.21.9791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dohlsten M., Hedlund G., Akerblom E., Lando P. A., Kalland T. Monoclonal antibody-targeted superantigens: a different class of anti-tumor agents. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9287–9291. doi: 10.1073/pnas.88.20.9287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dohlsten M., Hedlund G., Sjögren H. O., Carlsson R. Two subsets of human CD4+ T helper cells differing in kinetics and capacities to produce interleukin 2 and interferon-gamma can be defined by the Leu-18 and UCHL1 monoclonal antibodies. Eur J Immunol. 1988 Aug;18(8):1173–1178. doi: 10.1002/eji.1830180805. [DOI] [PubMed] [Google Scholar]
  10. Dohlsten M., Lando P. A., Björk P., Abrahmsén L., Ohlsson L., Lind P., Kalland T. Immunotherapy of human colon cancer by antibody-targeted superantigens. Cancer Immunol Immunother. 1995 Sep;41(3):162–168. doi: 10.1007/BF01521342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dohlsten M, Kalland T, Gunnarsson P, Antonsson P, Molander A, Olsson J, d'Argy R, Ohlsson L, Soegaard M, Persson R. Man-made superantigens: Tumor-selective agents for T-cell-based therapy. Adv Drug Deliv Rev. 1998 Apr 6;31(1-2):131–142. [PubMed] [Google Scholar]
  12. Forsberg G., Forsgren M., Jaki M., Norin M., Sterky C., Enhörning A., Larsson K., Ericsson M., Björk P. Identification of framework residues in a secreted recombinant antibody fragment that control production level and localization in Escherichia coli. J Biol Chem. 1997 May 9;272(19):12430–12436. doi: 10.1074/jbc.272.19.12430. [DOI] [PubMed] [Google Scholar]
  13. Garrido F., Ruiz-Cabello F., Cabrera T., Pérez-Villar J. J., López-Botet M., Duggan-Keen M., Stern P. L. Implications for immunosurveillance of altered HLA class I phenotypes in human tumours. Immunol Today. 1997 Feb;18(2):89–95. doi: 10.1016/s0167-5699(96)10075-x. [DOI] [PubMed] [Google Scholar]
  14. Goldenberg M. M. Trastuzumab, a recombinant DNA-derived humanized monoclonal antibody, a novel agent for the treatment of metastatic breast cancer. Clin Ther. 1999 Feb;21(2):309–318. doi: 10.1016/S0149-2918(00)88288-0. [DOI] [PubMed] [Google Scholar]
  15. Hansson J., Ohlsson L., Persson R., Andersson G., Ilbäck N. G., Litton M. J., Kalland T., Dohlsten M. Genetically engineered superantigens as tolerable antitumor agents. Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2489–2494. doi: 10.1073/pnas.94.6.2489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hedlund G., Dohlsten M., Lando P. A., Kalland T. Staphylococcal enterotoxins direct and trigger CTL killing of autologous HLA-DR+ mononuclear leukocytes and freshly prepared leukemia cells. Cell Immunol. 1990 Sep;129(2):426–434. doi: 10.1016/0008-8749(90)90218-g. [DOI] [PubMed] [Google Scholar]
  17. Hole N., Stern P. L. A 72 kD trophoblast glycoprotein defined by a monoclonal antibody. Br J Cancer. 1988 Mar;57(3):239–246. doi: 10.1038/bjc.1988.53. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hole N., Stern P. L. Isolation and characterization of 5T4, a tumour-associated antigen. Int J Cancer. 1990 Jan 15;45(1):179–184. doi: 10.1002/ijc.2910450132. [DOI] [PubMed] [Google Scholar]
  19. Hudson K. R., Tiedemann R. E., Urban R. G., Lowe S. C., Strominger J. L., Fraser J. D. Staphylococcal enterotoxin A has two cooperative binding sites on major histocompatibility complex class II. J Exp Med. 1995 Sep 1;182(3):711–720. doi: 10.1084/jem.182.3.711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hudson P. J. Recombinant antibody constructs in cancer therapy. Curr Opin Immunol. 1999 Oct;11(5):548–557. doi: 10.1016/s0952-7915(99)00013-8. [DOI] [PubMed] [Google Scholar]
  21. Hudson P. J. Recombinant antibody fragments. Curr Opin Biotechnol. 1998 Aug;9(4):395–402. doi: 10.1016/s0958-1669(98)80014-1. [DOI] [PubMed] [Google Scholar]
  22. Johnson H. M., Russell J. K., Pontzer C. H. Staphylococcal enterotoxin microbial superantigens. FASEB J. 1991 Sep;5(12):2706–2712. doi: 10.1096/fasebj.5.12.1916093. [DOI] [PubMed] [Google Scholar]
  23. Litton M. J., Dohlsten M., Lando P. A., Kalland T., Ohlsson L., Andersson J., Andersson U. Antibody-targeted superantigen therapy induces tumor-infiltrating lymphocytes, excessive cytokine production, and apoptosis in human colon carcinoma. Eur J Immunol. 1996 Jan;26(1):1–9. doi: 10.1002/eji.1830260102. [DOI] [PubMed] [Google Scholar]
  24. Massagué J. Purification of type-alpha transforming growth factor from transformed cells. Methods Enzymol. 1987;146:103–112. doi: 10.1016/s0076-6879(87)46013-8. [DOI] [PubMed] [Google Scholar]
  25. McLaughlin P., Grillo-López A. J., Link B. K., Levy R., Czuczman M. S., Williams M. E., Heyman M. R., Bence-Bruckler I., White C. A., Cabanillas F. Rituximab chimeric anti-CD20 monoclonal antibody therapy for relapsed indolent lymphoma: half of patients respond to a four-dose treatment program. J Clin Oncol. 1998 Aug;16(8):2825–2833. doi: 10.1200/JCO.1998.16.8.2825. [DOI] [PubMed] [Google Scholar]
  26. Modjtahedi H., Hickish T., Nicolson M., Moore J., Styles J., Eccles S., Jackson E., Salter J., Sloane J., Spencer L. Phase I trial and tumour localisation of the anti-EGFR monoclonal antibody ICR62 in head and neck or lung cancer. Br J Cancer. 1996 Jan;73(2):228–235. doi: 10.1038/bjc.1996.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mulder W. M., Stern P. L., Stukart M. J., de Windt E., Butzelaar R. M., Meijer S., Adér H. J., Claessen A. M., Vermorken J. B., Meijer C. J. Low intercellular adhesion molecule 1 and high 5T4 expression on tumor cells correlate with reduced disease-free survival in colorectal carcinoma patients. Clin Cancer Res. 1997 Nov;3(11):1923–1930. [PubMed] [Google Scholar]
  28. Myers K. A., Rahi-Saund V., Davison M. D., Young J. A., Cheater A. J., Stern P. L. Isolation of a cDNA encoding 5T4 oncofetal trophoblast glycoprotein. An antigen associated with metastasis contains leucine-rich repeats. J Biol Chem. 1994 Mar 25;269(12):9319–9324. [PubMed] [Google Scholar]
  29. Ortegel J. W., Staren E. D., Faber L. P., Warren W. H., Braun D. P. Cytokine biosynthesis by tumor-infiltrating T lymphocytes from human non-small-cell lung carcinoma. Cancer Immunol Immunother. 2000 Feb;48(11):627–634. doi: 10.1007/s002620050011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Persson B, Persson R, Weiner LM, Alpaugh RK. Overview of clinical trials employing antibody-targeted superantigens. Adv Drug Deliv Rev. 1998 Apr 6;31(1-2):143–152. doi: 10.1016/s0169-409x(97)00098-7. [DOI] [PubMed] [Google Scholar]
  31. Riethmüller G., Holz E., Schlimok G., Schmiegel W., Raab R., Höffken K., Gruber R., Funke I., Pichlmaier H., Hirche H. Monoclonal antibody therapy for resected Dukes' C colorectal cancer: seven-year outcome of a multicenter randomized trial. J Clin Oncol. 1998 May;16(5):1788–1794. doi: 10.1200/JCO.1998.16.5.1788. [DOI] [PubMed] [Google Scholar]
  32. Riethmüller G., Schneider-Gädicke E., Johnson J. P. Monoclonal antibodies in cancer therapy. Curr Opin Immunol. 1993 Oct;5(5):732–739. doi: 10.1016/0952-7915(93)90129-g. [DOI] [PubMed] [Google Scholar]
  33. Rosendahl A., Kristensson K., Carlsson M., Skartved N. J., Riesbeck K., Søgaard M., Dohlsten M. Long-term survival and complete cures of B16 melanoma-carrying animals after therapy with tumor-targeted IL-2 and SEA. Int J Cancer. 1999 Mar 31;81(1):156–163. doi: 10.1002/(sici)1097-0215(19990331)81:1<156::aid-ijc25>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
  34. Schad E. M., Zaitseva I., Zaitsev V. N., Dohlsten M., Kalland T., Schlievert P. M., Ohlendorf D. H., Svensson L. A. Crystal structure of the superantigen staphylococcal enterotoxin type A. EMBO J. 1995 Jul 17;14(14):3292–3301. doi: 10.1002/j.1460-2075.1995.tb07336.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Southall P. J., Boxer G. M., Bagshawe K. D., Hole N., Bromley M., Stern P. L. Immunohistological distribution of 5T4 antigen in normal and malignant tissues. Br J Cancer. 1990 Jan;61(1):89–95. doi: 10.1038/bjc.1990.20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Starzynska T., Marsh P. J., Schofield P. F., Roberts S. A., Myers K. A., Stern P. L. Prognostic significance of 5T4 oncofetal antigen expression in colorectal carcinoma. Br J Cancer. 1994 May;69(5):899–902. doi: 10.1038/bjc.1994.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Starzynska T., Rahi V., Stern P. L. The expression of 5T4 antigen in colorectal and gastric carcinoma. Br J Cancer. 1992 Nov;66(5):867–869. doi: 10.1038/bjc.1992.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Starzynska T., Wiechowska-Kozlowska A., Marlicz K., Bromley M., Roberts S. A., Lawniczak M., Kolodziej B., Zyluk A., Stern P. L. 5T4 oncofetal antigen in gastric carcinoma and its clinical significance. Eur J Gastroenterol Hepatol. 1998 Jun;10(6):479–484. doi: 10.1097/00042737-199806000-00008. [DOI] [PubMed] [Google Scholar]
  39. Søgaard M., Ohlsson L., Kristensson K., Rosendahl A., Sjoberg A., Forsberg G., Kalland T., Dohlsten M. Treatment with tumor-reactive Fab-IL-2 and Fab-staphylococcal enterotoxin A fusion proteins leads to sustained T cell activation, and long-term survival of mice with established tumors. Int J Oncol. 1999 Nov;15(5):873–882. doi: 10.3892/ijo.15.5.873. [DOI] [PubMed] [Google Scholar]
  40. Vater C. A., Reid K., Bartle L. M., Goldmacher V. S. Characterization of antibody binding to cell surface antigens using a plasma membrane-bound plate assay. Anal Biochem. 1995 Jan 1;224(1):39–50. doi: 10.1006/abio.1995.1006. [DOI] [PubMed] [Google Scholar]
  41. Winter G., Milstein C. Man-made antibodies. Nature. 1991 Jan 24;349(6307):293–299. doi: 10.1038/349293a0. [DOI] [PubMed] [Google Scholar]
  42. van de Griend R. J., Giphart M. J., Van Krimpen B. A., Bolhuis R. L. Human T cell clones exerting multiple cytolytic activities show heterogeneity in susceptibility to inhibition by monoclonal antibodies. J Immunol. 1984 Sep;133(3):1222–1229. [PubMed] [Google Scholar]
  43. van de Loosdrecht A. A., Nennie E., Ossenkoppele G. J., Beelen R. H., Langenhuijsen M. M. Cell mediated cytotoxicity against U 937 cells by human monocytes and macrophages in a modified colorimetric MTT assay. A methodological study. J Immunol Methods. 1991 Jul 26;141(1):15–22. doi: 10.1016/0022-1759(91)90205-t. [DOI] [PubMed] [Google Scholar]

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