Antibody-cytokine fusion proteins (“immunocytokines”) represent a promising class of armed antibody products, which allow the selective delivery of potent pro-inflammatory payloads at the tumor site. The antibody-based selective delivery of interleukin-2 (IL2) is particularly attractive for the treatment of metastatic melanoma, an indication for which this cytokine received marketing approval from the US Food and drug administration. We used the K1735M2 immunocompetent syngeneic model of murine melanoma to study the therapeutic activity of F8–IL2, an immunocytokine based on the F8 antibody in diabody format, fused to human IL2. F8–IL2 was shown to selectively localize at the tumor site in vivo, following intravenous administration, and to mediate tumor growth retardation, which was potentiated by the combination with paclitaxel or dacarbazine. Combination treatment led to a substantially more effective tumor growth inhibition, compared to the cytotoxic drugs used as single agents, without additional toxicity. Analysis of the immune infiltrate revealed a significant accumulation of CD4+ T cells 24 h after the administration of the combination. The fusion proteins F8–IL2 and L19–IL2, specific to the alternatively spliced extra domain A and extra domain B of fibronectin respectively, were also studied in combination with tumor necrosis factor (TNF)-based immunocytokines. The combination treatment was superior to the action of the individual immunocytokines and was able to eradicate neoplastic lesions after a single intratumoral injection, a procedure that is being clinically used for the treatment of Stage IIIC melanoma. Collectively, these data reinforce the rationale for the use of IL2-based immunocytokines in combination with cytotoxic agents or TNF-based immunotherapy for the treatment of melanoma patients.
This is a preview of subscription content, log in to check access.
The authors thank Kathrin Schwager for her help with biodistribution and immunofluorescence experiments. The study was supported by Swiss National Science Foundation, the Swiss Federal Institute of Technology (ETH), the European Union (FP7 ADAMANT and PRIAT Project) and the Kommission für Technologie und Innovation (KTI MedTech Award to Philochem).
Conflict of interest
Dario Neri is founder and shareholder of Philogen, a biotech company that owns the F8 antibody. Francesca Pretto and Giuliano Elia are Philochem employees. Nadia Castioni declares no conflict of interest.
Baluna R, Sausville EA, Stone MJ, Stetler-Stevenson MA, Uhr JW, Vitetta ES (1996) Decreases in levels of serum fibronectin predict the severity of vascular leak syndrome in patients treated with ricin a chain-containing immunotoxins. Clin Cancer Res 2(10):1705–1712PubMedGoogle Scholar
Schwartzentruber DJ (2001) Guidelines for the safe administration of high-dose interleukin-2. J Immunother 24(4):287–293PubMedCrossRefGoogle Scholar
Ko YJ, Bubley GJ, Weber R et al (2004) Safety, pharmacokinetics, and biological pharmacodynamics of the immunocytokine EMD 273066 (huKS-IL2): results of a phase I trial in patients with prostate cancer. J Immunother 27(3):232–239PubMedCrossRefGoogle Scholar
Carnemolla B, Borsi L, Balza E et al (2002) Enhancement of the antitumor properties of interleukin-2 by its targeted delivery to the tumor blood vessel extracellular matrix. Blood 99(5):1659–1665PubMedCrossRefGoogle Scholar
Villa A, Trachsel E, Kaspar M et al (2008) A high-affinity human monoclonal antibody specific to the alternatively spliced EDA domain of fibronectin efficiently targets tumor neo-vasculature in vivo. Int J Cancer 122(11):2405–2413. doi:10.1002/ijc.23408PubMedCrossRefGoogle Scholar
Tarli L, Balza E, Viti F et al (1999) A high-affinity human antibody that targets tumoral blood vessels. Blood 94(1):192–198PubMedGoogle Scholar
Schwager K, Hemmerle T, Aebischer D, Neri D (2013) The immunocytokine L19–IL2 eradicates cancer when used in combination with CTLA-4 blockade or with L19–TNF. J Invest Dermatol 133(3):751–758. doi:10.1038/jid.2012.376PubMedCrossRefGoogle Scholar
Halin C, Gafner V, Villani ME et al (2003) Synergistic therapeutic effects of a tumor targeting antibody fragment, fused to interleukin 12 and to tumor necrosis factor alpha. Cancer Res 63(12):3202–3210PubMedGoogle Scholar
Schwager K, Kaspar M, Bootz F et al (2009) Preclinical characterization of DEKAVIL (F8-IL10), a novel clinical-stage immunocytokine which inhibits the progression of collagen-induced arthritis. Arthritis Res Ther 11(5):R142. doi:10.1186/ar2814PubMedCentralPubMedCrossRefGoogle Scholar
Johannsen M, Spitaleri G, Curigliano G et al (2010) The tumour-targeting human L19–IL2 immunocytokine: preclinical safety studies, phase I clinical trial in patients with solid tumours and expansion into patients with advanced renal cell carcinoma. Eur J Cancer 46(16):2926–2935. doi:10.1016/j.ejca.2010.07.033PubMedCrossRefGoogle Scholar
Eigentler TK, Weide B, de Braud F et al (2011) A dose-escalation and signal-generating study of the immunocytokine L19–IL2 in combination with dacarbazine for the therapy of patients with metastatic melanoma. Clin Cancer Res 17(24):7732–7742. doi:10.1158/1078-0432.CCR-11-1203PubMedCrossRefGoogle Scholar
Spitaleri G, Berardi R, Pierantoni C et al (2013) Phase I/II study of the tumour-targeting human monoclonal antibody-cytokine fusion protein L19–TNF in patients with advanced solid tumours. J Cancer Res Clin Oncol 139(3):447–455. doi:10.1007/s00432-012-1327-7PubMedGoogle Scholar
Papadia F, Basso V, Patuzzo R et al (2013) Isolated limb perfusion with the tumor-targeting human monoclonal antibody-cytokine fusion protein L19–TNF plus melphalan and mild hyperthermia in patients with locally advanced extremity melanoma. J Surg Oncol 107(2):173–179. doi:10.1002/jso.23168PubMedCrossRefGoogle Scholar
Weide B, Eigentler TK, Pflugfelder A et al (2011) Survival after intratumoral interleukin-2 treatment of 72 melanoma patients and response upon the first chemotherapy during follow-up. Cancer Immunol Immunother 60(4):487–493. doi:10.1007/s00262-010-0957-3PubMedCrossRefGoogle Scholar
Weide B, Derhovanessian E, Pflugfelder A et al (2010) High response rate after intratumoral treatment with interleukin-2: results from a phase 2 study in 51 patients with metastasized melanoma. Cancer 116(17):4139–4146. doi:10.1002/cncr.25156PubMedCrossRefGoogle Scholar
Weide B, Eigentler TK, Pflugfelder A et al (2014) Intralesional treatment of stage III metastatic melanoma patients with L19–IL2 results in sustained clinical and systemic immunologic responses. Cancer Immunol Res. doi:10.1158/2326-6066.CIR-13-0206PubMedGoogle Scholar
Kripke ML (1979) Speculations on the role of ultraviolet radiation in the development of malignant melanoma. J Natl Cancer Inst 63(3):541–548PubMedGoogle Scholar
Talmadge JE, Fidler IJ (1982) Enhanced metastatic potential of tumor cells harvested from spontaneous metastases of heterogeneous murine tumors. J Natl Cancer Inst 69(4):975–980PubMedGoogle Scholar
Frey K, Schliemann C, Schwager K, Giavazzi R, Johannsen M, Neri D (2010) The immunocytokine F8–IL2 improves the therapeutic performance of sunitinib in a mouse model of renal cell carcinoma. J Urol 184(6):2540–2548. doi:10.1016/j.juro.2010.07.030PubMedCrossRefGoogle Scholar
Hemmerle T, Probst P, Giovannoni L, Green AJ, Meyer T, Neri D (2013) The antibody-based targeted delivery of TNF in combination with doxorubicin eradicates sarcomas in mice and confers protective immunity. Br J Cancer 109(5):1206–1213. doi:10.1038/bjc.2013.421PubMedCrossRefGoogle Scholar
Pasche N, Wulhfard S, Pretto F, Carugati E, Neri D (2012) The antibody-based delivery of interleukin-12 to the tumor neovasculature eradicates murine models of cancer in combination with paclitaxel. Clin Cancer Res 18(15):4092–4103. doi:10.1158/1078-0432.CCR-12-0282PubMedCrossRefGoogle Scholar
Schwager K, Bootz F, Imesch P, Kaspar M, Trachsel E, Neri D (2011) The antibody-mediated targeted delivery of interleukin-10 inhibits endometriosis in a syngeneic mouse model. Hum Reprod 26(9):2344–2352. doi:10.1093/humrep/der195PubMedCrossRefGoogle Scholar
Gutbrodt KL, Schliemann C, Giovannoni L et al. (2013) Antibody-based delivery of interleukin-2 to neovasculature has potent activity against acute myeloid leukemia. Sci Transl Med 5 (201):201ra118. doi 10.1126/scitranslmed.3006221
Santimaria M, Moscatelli G, Viale GL et al (2003) Immunoscintigraphic detection of the ED-B domain of fibronectin, a marker of angiogenesis, in patients with cancer. Clin Cancer Res 9(2):571–579PubMedGoogle Scholar
Sauer S, Erba PA, Petrini M et al (2009) Expression of the oncofetal ED-B-containing fibronectin isoform in hematologic tumors enables ED-B-targeted 131I-L19SIP radioimmunotherapy in hodgkin lymphoma patients. Blood 113(10):2265–2274. doi:10.1182/blood-2008-06-160416PubMedCrossRefGoogle Scholar
Hemmerle T, Neri D (2014) The antibody-based targeted delivery of interleukin-4 and 12 to the tumor neovasculature eradicates tumors in three mouse models of cancer. Int J Cancer 134(2):467–477. doi:10.1002/ijc.28359PubMedCrossRefGoogle Scholar