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 10–10 M, 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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References
Abrahmsén L, Dohlsten M, Segren S, Björk P, Jonsson E and Kalland T (1995) Characterization of two distinct MHC class II binding sites in the superantigen staphylococcal enterotoxin A. EMBO J 14: 2978–2986
Alpaugh RK, Weiner LM, Persson R and Persson B (1998) Overview of clinical trials employing antibody-targeted superantigens. Adv Drug Deliv Rev 31: 143–152
Bontkes HJ, van Duin M, de Gruijl TD, Duggan-Keen MF, Walboomers JM, Stukart MJ, Verheijen RH, Helmerhorst TJ, Meijer CJ, Scheper RJ, Stevens FR, Dyer PA, Sinnott P and Stern PL (1998) HPV 16 infection and progression of cervical intra-epithelial neoplasia: analysis of HLA polymorphism and HPV 16 E6 sequence variants. Int J Cancer 78: 166–171
Bontkes HJ, de Gruijl TD, Bijl A, Verheijen RH, Meijer CJ, Scheper RJ, Stern PL, Burns JE, Maitland NJ and Walboomers JM (1999) Human papillomavirus type 16 E2-specific T-helper lymphocyte responses in patients with cervical intraepithelial neoplasia. J Gen Virol 80: 2453–2459
Brodin TN, Persson R, Soegaard M, Ohlsson L, d’Argy R, Olsson J, Molander A, Antonsson P, Gunnarsson P-O, Kalland T and Dohlsten M (1998) Man-made superantigens: Tumour selective agents for T-cell-based therapy. Adv Drug Deliv Rev 31: 131–142
Carsberg CJ, Myers KA, Evans GS, Allen TD and Stern PL (1995). Metastasis-associated 5T4 oncofoetal antigen is concentrated at microvillus projections of the plasma membrane, J Cell Sci 2905–2916
Carsberg CJ, Myers KA and Stern PL (1996) Metastasis-associated 5T4 antigen disrupts cell-cell contacts and induces cellular motility in epithelial cells. Int J Cancer 27: 84–92
Dall’Acqua W and Carter P (1998) Antibody engineering. Curr Opin Struct Biol 8: 443–450
Dohlsten M, Hedlund G, Sjögren HO and Carlsson R (1998). 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 1173–1178
Dohlsten M, Hedlund G, Åkerblom E, Lando P and Kalland T (1991) Monoclonal antibody-targeted superantigens: a different class of anti-tumour agents. Proc Natl Acad Sci USA 88: 9287–9281
Dohlsten M, Bjorklund M, Sundstedt A, Hedlund G, Samson D and Kalland T (1993) Immunopharmacology of the superantigen Staphylococcal enterotoxin A in T-cell receptor Vb3 transgenic mice. Immunology 79: 520–527
Dohlsten M, Abrahmsén L, Björk P, Lando PA, Hedlund G, Forsberg G, Brodin T, Gascoigne NRJ, Förberg C, Lind P and Kalland T (1994) Monoclonal antibody-superantigen fusion proteins: Tumour specific agents for T cell based tumour therapy. Proc Natl Acad Sci 91: 8945–8949
Dohlsten M, Hansson J, Ohlsson L, Litton M and Kalland T (1995a) Antibody targeted superantigens are potent inducers of tumour-infiltrating T lymphocytes in vivo. Proc Natl Acad Sci USA 92: 9791–9795
Dohlsten M, Lando PA, Björk P, Abrahmsén L, Ohlsson L, Lind P and Kalland T (1995b) Immuno-therapy of human colon cancer by antibody targeted superantigens. Cancer Immunol. Immunotherapy 41: 162–168
Forsberg G, Forsgren M, Jaki M, Norin M, Sterky C, Enhörning Å, Larsson K, Ericsson M and Björk P (1997) Identification of framework residues in a secreted recombinant antibody fragment that control production level and localization in Escherichia coli. J Biol Chem 272: 12430–12436
Garrido F, Ruiz-Cabello F, Cabrera T, Perez-Villar JJ, Lopez-Botet M, Duggan-Keen M and Stern PL (1997) Implications for immunosurveillance of altered HLA class I phenotypes in human tumours. Immunol Today 18: 89–95
Goldenberg MM (1999) Traztusumab, a recombinant DNA-derived humanized monoclonal antibody, a novel agent for the treatment of metastatic breast cancer. Clin Ther 21: 309–318
Hansson J, Ohlsson L, Persson R, Andersson G, Ilbäck N-G, Liton M, Kalland T and Dohlsten M (1997) Genetically engineered superantigens as tolerable antitumor agents. Proc Natl Acad Sci USA 94: 2489–2494
Hedlund G, Dohlsten M, Lando PA and Kalland T (1990) Staphylococcal enterotoxins direct and trigger CTL killing of autologous HLA-DR+ mononuclear leukocytes and freshly prepared leukemia cells. Cell Immunol 129: 426–434
Hole N and Stern PL (1988) A 72 KD trophoblast glycoprotein defined by a monoclonal antibody. Br J Cancer 57: 239–246
Hole N and Stern PL (1990) Isolation and characterization of 5T4, a tumour-associated antigen. Int J Cancer 15: 179–184
Hudson KR, Tiedemann RE, Urban RG, Lowe SC, Strominger JL and Fraser JD (1995) Staphylococcal enterotoxin A has two cooperative binding sites on major histocompatibility complex class II. J Exp Med 182: 711–720
Hudson PJ (1998) Recombinant antibody fragments. Curr Opin Biotechnol 9: 395–402
Hudson PJ (1999) Recombinant antibody constructs in cancer therapy. Curr Opin Immunol 11: 548–557
Johnson HM, Russell JK and Pontzer CH (1991) Staphylococcal enterotoxin microbial superantigens. FASEB J 5: 2706–2712
Litton MJ, Dohlsten M, Lando PA, Kalland T, Ohlsson L, Andersson J and Andersson U (1996) Antibody-targeted superantigen therapy induces tumour-infiltrating lymphocytes, excessive cytokine production, and apoptosis in human colon carcinoma. Eur J Immunol 26: 1–9
Massague J (1987). Methods Enzymol 146: 103–
McLaughlin P, Grillo-Lopez AJ, Link BK, Levy R, Czuczman MS, Williams ME, Heyman MR, Bence-Bruckler I, White CA, Cabanillas F, Jain V, Ho AD, Lister J, Wey K, Shen D and Dallaire BK (1998) Rituximab chimeric anti-CD20 monoclonal antibody therapy for relapsed indolent lymphoma: half of patients respond to a four-dose treatment program. J Clin Oncol 16: 2825–2833
Modjtahedi H, Hickish T, Nicolson M, Moore J, Styles J, Eccles S, Jackson E, Salter J, Sloane J, Spencer L, Priest K, Smith I, Dean C and Gore M (1996) Phase I trial and tumour localisation of the anti-EGFR monoclonal antibody ICR62 in head and neck or lung cancer. Br J Cancer 73: 228–235
Mulder WM, Stern PL, Stukart MJ, de Windt E, Butzelaar RM, Meijer S, Ader HJ, Claessen AM, Vermorken JB, Meijer CJ, Wagstaff J, Scheper RJ and Bloemena E (1997) 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 3: 1923–1930
Myers KA, Rahi-Saund V, Davison MD, Young JA, Cheater AJ and Stern PL (1994) Isolation of a cDNA encoding 5T4 oncofetal trophoblast glycoprotein. An antigen associated with metastasis contains leucine-rich repeats. J Biol Chem 269: 9319–9324
Ortegel JW, Staren ED, Faber LP, Warren WH and Braun DP (2000) Cytokine biosynthesis by tumor infiltrating T lymphocytes from human non small-cell lung carcinoma. Cancer Immunol Immunother 48: 627–634
Riethmüller G, Schneider-Gaedicke E and Johnson JP (1993) Monoclonal antibodies in cancer therapy. Curr Opin Immunol 5: 732–739
Riethmuller G, Holz E, Schlimok G, Schmiegel W, Raab R, Hoffken K, Gruber R, Funke I, Pichlmaier H, Hirche H, Buggisch P, Witte J and Pichlmayr R (1998) Monoclonal antibody therapy for resected Dukes’ C colorectal cancer: seven-year outcome of a multicenter randomized trial. J Clin Oncol 16: 1788–1794
Rosendahl A, Kristensson K, Carlsson M, Skartved NJ, Riesbeck K, Soegaard M and Dohlsten M (1999) Long-term survival and complete cures of B16-melanoma carrying animals after therapy with tumour targeted IL-2 and SEA. Int J Cancer 81: 151–163
Scatchard G (1949) The attraction of proteins for small molecules and ions. Ann NY Acad Sci 51: 660–672
Schad EM, Zaitseva I, Zaitsev VN, Dohlsten M, Kalland T, Schlievert DH, Ohlendorf DH and Svensson LA (1995) Crystal structure of the superantigen staphylococcal enterotoxin type A. EMBO J 14: 3292–3301
Soegaard M, Ohlsson L, Kristensson K, Rosendahl A, Sjöberg A, Forsberg G, Kalland T and Dohlsten M (1999) Treatment with tumour-reactive Fab-IL-2 and Fab-Staphylococcal enteroxin A fusion proteins leads to sustained T cell activation and long-term survival of mice with established tumours. Int J Oncol 15: 873–882
Southall PJ, Boxer GM, Bagshawe KD, Hole N, Bromley M and Stern PL (1990) Immunohistological distribution of 5T4 antigen in normal and malignant tissues. Br J Cancer 61: 89–95
Starzynska T, Rahi V and Stern PL (1992) The expression of 5T4 antigen in colorectal and gastric carcinoma. Br J Cancer 66: 867–869
Starzynska T, Marsh PJ, Schofield PF, Roberts SA, Myers KA and Stern PL (1994) Prognostic significance of 5T4 oncofetal antigen expression in colorectal carcinoma. Br J Cancer 69: 899–902
Starzynska T, Wiechowska-Kozlowska A, Marlicz K, Bromley M, Roberts SA, Lawniczak M, Kolodziej B, Zyluk A and Stern PL (1998) 5T4 oncofetal antigen in gastric carcinoma and its clinical significance. Eur J Gastroenterol Hepatol 10: 479–484
Van De Griend RJ, Girhart MJ, Van Krimpen BA and Bolhuis RLH (1984) Human T cell clones exerting multiple cytolytic activities show heterogeneity in susceptibility to inhibition by monoclonal antibodies. J Immunol 133: 1222–1229
van de Loosdrecht AA, Nennie E, Ossenkoppele GJ, Beelen RH and Langenhuijsen MM (1991) Cell mediated cytotoxicity against U 937 cells by human monocytes and macrophages in a modified colorimetric MTT assay. A methodological study. J Immunol Methods 141: 15–22
Vater CA, Reid K, Bartle LM and Goldmacher VS (1995) Characterization of antibody binding to cell surface antigens using plasma membrane-bound plate assay. Anal Biochem 224: 39–50
Winter G and Milstein C (1991) Man-made antibodies. Nature 349: 293–299
Zimmermann S, Wels W, Froesch BA, Gerstmayer B, Stahel RA and Zangemeister-Wittke U 1997 A novel immunotoxin recognising the epithelial glycoprotein-2 has potent antitumoural activity on chemotherapy-resistant lung cancer. Cancer Immunol Immunother 44: 1–9
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Forsberg, G., Ohlsson, L., Brodin, T. et al. Therapy of human non-small-cell lung carcinoma using antibody targeting of a modified superantigen. Br J Cancer 85, 129–136 (2001). https://doi.org/10.1054/bjoc.2001.1891
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DOI: https://doi.org/10.1054/bjoc.2001.1891
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