To investigate the antitumor activities of an anti-ErbB2 scFv-Fc-interleukin 2 (IL-2) fusion protein (HFI) in vitro and in vivo.
Fusion protein HFI was constructed. The efficacy of HFI in mediating tumor cell lysis was determined by colorimetric lactate dehydrogenase release assays. The antitumor activity of HFI was evaluated in tumor xenograft models.
The fusion protein was folded as a homodimer formed by covalently linking Fc portions and it retained ErbB2 specificity and IL-2 biological activity. HFI mediated antibody-dependent cell-mediated cytotoxicity (ADCC) at low effector-to-target ratios in vitro and improved the therapeutic efficacy of IL-2 in experiments in vivo.
The genetically-engineered anti-ErbB2 scFv-Fc-IL-2 fusion protein exhibited high efficiency both in mediating ADCC in vitro and significant antitumor activity in tumor xenograft models.
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Hank JA, Robinson RR, Surfus J, Mueller BM, Reisfeld RA, Cheung NK, et al. Augmentation of antibody dependent cell mediated cytotoxicity following in vivo therapy with recombinant interleukin 2. Cancer Res 1990; 50: 5234–9.
Rosenberg SA . Progress in human tumour immunology and immunotherapy. Nature 2001; 411: 380–4.
Rosenberg SA, Lotze MT, Yang JC, Aebersold PM, Linehan WM, Seipp CA, et al. Experience with the use of high-dose interleukin-2 in the treatment of 652 cancer patients. Ann Surg 1989; 210: 474–84.
Rosenberg SA, Packard BS, Aebersold PM, Solomon D, Topalian SL, Toy ST, et al. Use of tumor-infiltrating lymphocytes and interleukin-2 in the immunotherapy of patients with metastatic melanoma. A preliminary report. N Engl J Med 1988; 319: 1676–80.
Rosenberg SA, Yannelli JR, Yang JC, Topalian SL, Schwartzentruber DJ, Weber JS, et al. Treatment of patients with metastatic melanoma with autologous tumor-infiltrating lymphocytes and interleukin 2. J Natl Cancer Inst 1994; 86: 1159–66.
Roth JA, Cristiano RJ . Gene therapy for cancer: What have we done and where are we going? J Natl Cancer Inst 1997; 89: 21–39.
Zatloukal K, Schneeberger A, Berger M, Schmidt W, Koszik F, Kutil R, et al. Elicitation of a systemic and protective anti-melanoma immune response by an IL-2-based vaccine. Assessment of critical cellular and molecular parameters. J Immunol 1995; 154: 3406–19.
Dranoff G, Mulligan RC . Gene transfer as cancer therapy. Adv Immunol 1995; 58: 417–54.
Maas RA, Dullens HF, Den OW . Interleukin-2 in cancer treatment: disappointing or (still) promising?. A review. Cancer Immunol Immunother 1993; 36: 141–8.
Cortesina G, De SA, Giovarelli M, Barioglio MG, Cavallo GP, Jemma C, et al. Treatment of recurrent squamous cell carcinoma of the head and neck with low doses of interleukin-2 injected perilymphatically. Cancer 1988; 62: 2482–5.
Forni G, Musso T, Santoni A, Giovarelli M . Local administration of interleukin-2 activates lymphocytes from tumor bearing mice to recruit host immunoreactivity and inhibit tumor growth. Prog Clin Biol Res 1987; 244: 105–14.
Pizza G, Severini G, Menniti D, De VC, Corrado F . Tumour regression after intralesional injection of interleukin 2 (IL-2) in bladder cancer. Preliminary report. Int J Cancer 1984; 34: 359–67.
Rutten VP, Klein WR, De Jong WA, Misdorp W, Den OW, Steerenberg PA, et al. Local interleukin-2 therapy in bovine ocular squamous cell carcinoma. A pilot study. Cancer Immunol Immunother 1989; 30: 165–9.
Hurford RK Jr, Dranoff G, Mulligan RC, Tepper RI . Gene therapy of metastatic cancer by in vivo retroviral gene targeting. Nat Genet 1995; 10: 430–5.
Schmidt W, Schweighoffer T, Herbst E, Maass G, Berger M, Schilcher F, et al. Cancer vaccines: the interleukin 2 dosage effect. Proc Natl Acad Sci USA 1995; 92: 4711–4.
Soiffer R, Lynch T, Mihm M, Jung K, Rhuda C, Schmollinger JC, et al. Vaccination with irradiated autologous melanoma cells engineered to secrete human granulocyte-macrophage colony-stimulating factor generates potent antitumor immunity in patients with metastatic melanoma. Proc Natl Acad Sci USA 1998; 95: 13 141–6.
Gillies SD, Lan Y, Wesolowski JS, Qian X, Reisfeld RA, Holden S, et al. Antibody-IL-12 fusion proteins are effective in scid mouse models of prostate and colon carcinoma metastases. J Immunol 1998; 160: 6195–203.
Gillies SD, Reilly EB, Lo KM, Reisfeld RA . Antibody-targeted interleukin 2 stimulates t-cell killing of autologous tumor cells. Proc Natl Acad Sci USA 1992; 89: 1428–32.
Penichet ML, Morrison SL . Antibody-cytokine fusion proteins for the therapy of cancer. J Immunol Methods 2001; 248: 91–101.
Lode HN, Xiang R, Dreier T, Varki NM, Gillies SD, Reisfeld RA . Natural killer cell-mediated eradication of neuroblastoma metastases to bone marrow by targeted interleukin-2 therapy. Blood 1998; 91: 1706–15.
Hornick JL, Khawli LA, Hu P, Lynch M, Anderson PM, Epstein AL . Chimeric CLL-1 antibody fusion proteins containing granulocyte-macrophage colony-stimulating factor or interleukin-2 with specificity for beta-cell malignancies exhibit enhanced effector functions while retaining tumor targeting properties. Blood 1997; 89: 4437–47.
Xiang R, Lode HN, Dolman CS, Dreier T, Varki NM, Qian X, et al. Elimination of established murine colon carcinoma metastases by antibody-interleukin 2 fusion protein therapy. Cancer Res 1997; 57: 4948–55.
Molina MA, Codony-Servat J, Albanell J, Rojo F, Arribas J, Baselga J . Trastuzumab (herceptin), a humanized anti-her2 receptor monoclonal antibody, inhibits basal and activated her2 ectodomain cleavage in breast cancer cells. Cancer Res 2001; 61: 4744–9.
Shak S . Overview of the trastuzumab (herceptin) anti-her2 monoclonal antibody clinical program in her2-overexpressing meta-static breast cancer. Herceptin Multinational Investigator Study Group. Semin Oncol 1999; 26: 71–7.
Yip YL, Ward RL . Anti-erbb-2 monoclonal antibodies and erbb-2-directed vaccines. Cancer Immunol Immunother 2002; 50: 569–87.
Shi M, Xie Z, Feng J, Sun Y, Yu M, Shen B, et al. A recombinant anti-erbb2, scfv-fc-il-2 fusion protein retains antigen specificity and cytokine function. Biotechnol Lett 2003; 25: 815–9.
Shi M, Xie Z, Yu M, Shen B, Guo N . Controlled growth of Chinese hamster ovary cells and high expression of antibody-IL-2 fusion proteins by temperature manipulation. Biotechnol Lett 2005; 27: 1879–84.
Emlet DR, Schwartz R, Brown KA, Pollice AA, Smith CA, Shackney SE . Her2 expression as a potential marker for response to therapy targeted to the egfr. Br J Cancer 2006; 94: 1144–53.
Dean GS, Pusztai L, Xu FJ, O'Briant K, DeSombre K, Conaway M, et al. Cell surface density of p185(c-erbb-2) determines susceptibility to anti-p185(c-erbb-2)-ricin a chain (rta) immunotoxin therapy alone and in combination with anti-p170(egfr)-rta in ovarian cancer cells. Clin Cancer Res 1998; 4: 2545–50.
Xu F, Yu Y, Le XF, Boyer C, Mills GB, Bast RC Jr . The outcome of heregulin-induced activation of ovarian cancer cells depends on the relative levels of her-2 and her-3 expression. Clin Cancer Res 1999; 5: 3653–60.
Barr TA, McCormick AL, Carlring J, Heath AW . A potent adjuvant effect of CD40 antibody attached to antigen. Immunology 2003; 109: 87–92.
Halin C, Rondini S, Nilsson F, Berndt A, Kosmehl H, Zardi L, et al. Enhancement of the antitumor activity of interleukin-12 by targeted delivery to neovasculature. Nat Biotechnol 2002; 20: 264–9.
Grimm EA, Mazumder A, Zhang HZ, Rosenberg SA . Lymphokine-activated killer cell phenomenon. Lysis of natural killer-resistant fresh solid tumor cells by interleukin 2-activated autologous human peripheral blood lymphocytes. J Exp Med 1982; 155: 1823–41.
Hayes RL, Koslow M, Hiesiger EM, Hymes KB, Hochster HS, Moore EJ, et al. Improved long term survival after intracavitary interleukin-2 and lymphokine-activated killer cells for adults with recurrent malignant glioma. Cancer 1995; 76: 840–52.
Maass G, Schmidt W, Berger M, Schilcher F, Koszik F, Schneeberger A, et al. Priming of tumor-specific t cells in the draining lymph nodes after immunization with interleukin 2-secreting tumor cells: three consecutive stages may be required for successful tumor vaccination. Proc Natl Acad Sci USA 1995; 92: 5540–4.
Bukowski JF, Warner JF, Dennert G, Welsh RM . Adoptive transfer studies demonstrating the antiviral effect of natural killer cells in vivo. J Exp Med 1985; 161: 40–52.
Favrot MC, Floret D, Negrier S, Cochat P, Bouffet E, Zhou DC, et al. Systemic interleukin-2 therapy in children with progressive neuroblastoma after high dose chemotherapy and bone marrow transplantation. Bone Marrow Transplant 1989; 4: 499–503.
Koehne G, Zeller W, Stockschlaeder M, Zander AR . Phenotype of lymphocyte subsets after autologous peripheral blood stem cell transplantation. Bone Marrow Transplant 1997; 19: 149–56.
Uharek L, Zeis M, Glass B, Steinmann J, Dreger P, Gassmann W, et al. High lytic activity against human leukemia cells after activation of allogeneic NK cells by IL-12 and IL-2. Leukemia 1996; 10: 1758–64.
Nagayama H, Takahashi S, Takahashi T, Ogami K, Ikebuchi K, Tojo A, et al. Il-2/lak therapy for refractory acute monoblastic leukemia relapsing after unrelated allogeneic bone marrow transplantation. Bone Marrow Transplant 1999; 23: 183–5.
Lustgarten J . Anti-her-2/neu-il-2 or heregulin-il-2 fusions proteins redirect non-tumor specific ctl to the tumor site for tumor eradication. Cancer Immunol Immunother 2003; 52: 751–60.
Yu D, Hung MC . Role of erbb2 in breast cancer chemosensitivity. Bioessays 2000; 22: 673–80.
Yu D, Jing T, Liu B, Yao J, Tan M, McDonnell TJ, et al. Overexpression of erbb2 blocks taxol-induced apoptosis by up-regulation of p21cip1, which inhibits p34cdc2 kinase. Mol Cell 1998; 2: 581–91.
Yu D, Liu B, Jing T, Sun D, Price JE, Singletary SE, et al. Overexpression of both p185c-erbb2 and p170mdr-1 renders breast cancer cells highly resistant to taxol. Oncogene 1998; 16: 2087–94.
Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL . Human breast cancer: correlation of relapse and survival with amplification of the her-2/neu oncogene. Science 1987; 235: 177–82.
Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE, et al. Studies of the her-2/neu proto-oncogene in human breast and ovarian cancer. Science 1989; 244: 707–12.
Guan H, Jia SF, Zhou Z, Stewart J, Kleinerman ES . Herceptin down-regulates her-2/neu and vascular endothelial growth factor expression and enhances taxol-induced cytotoxicity of human ewing's sarcoma cells in vitro and in vivo. Clin Cancer Res 2005; 11: 2008–17.
Klapper LN, Waterman H, Sela M, Yarden Y . Tumor-inhibitory antibodies to her-2/erbb-2 may act by recruiting c-cbl and enhancing ubiquitination of her-2. Cancer Res 2000; 60: 3384–8.
Spiridon CI, Ghetie MA, Uhr J, Marches R, Li JL, Shen GL, et al. Targeting multiple her-2 epitopes with monoclonal antibodies results in improved antigrowth activity of a human breast cancer cell line in vitro and in vivo. Clin Cancer Res 2002; 8: 1720–30.
Baselga J, Norton L, Albanell J, Kim YM, Mendelsohn J . Recombinant humanized anti-her2 antibody (herceptin) enhances the antitumor activity of paclitaxel and doxorubicin against her2/neu overexpressing human breast cancer xenografts. Cancer Res 1998; 58: 2825–31.
Kim R, Tanabe K, Emi M, Uchida Y, Toge T . Modulation of tamoxifen sensitivity by antisense bc1-2 and trastuzumab in breast carcinoma cells. Cancer 2005; 103: 2199–207.
Lee S, Yang W, Lan KH, Sellappan S, Klos K, Hortobagyi G, et al. Enhanced sensitization to taxol-induced apoptosis by herceptin pretreatment in erbb2-overexpressing breast cancer cells. Cancer Res 2002; 62: 5703–10.
Pegram MD, Lipton A, Hayes DF, Weber BL, Baselga JM, Tripathy D, et al. Phase II study of receptor-enhanced chemosensitivity using recombinant humanized anti-p185her2/neu monoclonal antibody plus cisplatin in patients with her2/neu-overexpressing metastatic breast cancer refractory to chemotherapy treatment. J Clin Oncol 1998; 16: 2659–71.
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