Elsevier

The Journal of Urology

Volume 184, Issue 6, December 2010, Pages 2540-2548
The Journal of Urology

Investigative Urology
The Immunocytokine F8-IL2 Improves the Therapeutic Performance of Sunitinib in a Mouse Model of Renal Cell Carcinoma

https://doi.org/10.1016/j.juro.2010.07.030Get rights and content

Purpose

We investigated the therapeutic action of F8-IL2, a fusion protein consisting of the F8 antibody specific to the alternatively spliced extradomain-A of fibronectin, in diabody format and of human interleukin-2 in the Caki-1 (ATCC®) model of human renal cell carcinoma grafted subcutaneously in nude mice.

Materials and Methods

F8-IL2 was cloned, expressed in CHO cells and purified to homogeneity. This immunocytokine was administered alone or combined with 3 standard drugs commonly used as therapy for kidney cancer, including sunitinib, sorafenib and interferon-α, in 2 sets of doses and treatment schedules.

Results

Neither F8-IL2 nor any other therapeutic agent cured tumor bearing mice when used as a single agent. The best therapeutic results were observed for the combination of sunitinib with F8-IL2 in a continuous administration schedule, which yielded a 28% cure rate and substantial tumor growth retardation.

Conclusions

Considering that recombinant interleukin-2 based immunocytokines are now being investigated in several clinical trials in patients with cancer alone or combined with chemotherapy our preclinical results provide a motivation to study F8-IL2 combined with sunitinib in clinical trials in patients with kidney cancer.

Section snippets

Therapeutic Agents, Cell Lines and Animals

Human monoclonal antibodies in SIP format were previously described.16, 18, 19 We used L19-IL2.11 Human IFN-α-2b (Intron® A) was purchased as a 106 IU ampoule. We also used recombinant murine IFN-α-1 (PBL InterferonSource, Piscataway, New Jersey) (105 U). The cell lines Caki-1, RXF393, A498 (DCTD Tumor Repository, Charles River Laboratories, National Cancer Institute at Frederick, Frederick, Maryland), SN12K120 and CTLL-2 (ATCC) were grown according to vendor instructions. Female BALB/c nude

F8-IL2 Cloning, Expression and In Vitro Characterization

The F8-IL2 gene was assembled from the F8 antibody in diabody format,16, 17 which consists of the variable heavy and light domains joined together by a 5-amino acid polypeptide linker, and which was sequentially fused with the human IL2 gene.11, 12 F8-IL2 was cloned into the commercial pcDNA3.1 vector, which was then used to stably transfect CHO cells (fig. 1,A). The diabody format of F8 confers bivalent antigen binding to the immunocytokine (fig. 1, B), which was previously investigated in

Discussion

We report that the fully human immunocytokine F8-IL2 selectively localizes to kidney tumors implanted in subcutaneous and orthotopic mouse models, and this biopharmaceutical delays tumor growth. The therapeutic effect of F8-IL2 was potentiated by combination with sunitinib but not with sorafenib and IFN-α (fig. 3). In extensive therapy studies we used subcutaneously grafted Caki-1 tumors since facile implantation was compatible with the large number of animals needed and these slowly growing

Conclusions

When combined with other therapeutic agents, IL2 based immunocytokines could eradicate established lymphoma14 and neuroblastoma.30 F8-IL2 appears to be a promising new biopharmaceutical agent for RCC since it substantially delays tumor growth when used alone or combined with other agents. However, the fact that complete cure was only rarely achieved even in the most effective experimental setting (F8-IL2 plus sunitinib) motivates us to continue our investigations of novel therapeutic regimens

Acknowledgments

Valentina Scarlato, Mario Negri Institute, Milano, Italy, assisted with the orthotopic RCC mouse models. Dr. Camilla Bacci, Philogen, Siena, Italy, provided the L19-IL2 and huIL2 NIBSC standard. Dr. H. H. Fiebig, Freiburg, Germany, provided RXF393.

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    • Cited by (0)

    Study received approval from Veterinäramt des Kantons Zürich, Switzerland.

    Supported by the Swiss National Science Foundation, ETH Zürich, European Union ADAMANT and IMMUNO-PDT Projects, Swiss Cancer League, Swiss-Bridge Foundation, Stammbach Foundation, Italian Association for Cancer Research and Fondazione Cariplo 2008-2264.

    ⁎

    Financial interest and/or other relationship with Pfizer Germany, Bayer, Novartis Germany, Wyeth Germany, Roche Germany and GlaxoSmithKline.

    ‡

    Financial interest and/or other relationship with Philogen.

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