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Synergistic therapeutic effects of a tumor targeting antibody fragment, fused to interleukin 12 and to Tumor Necrosis Factor α

TitleSynergistic therapeutic effects of a tumor targeting antibody fragment, fused to interleukin 12 and to Tumor Necrosis Factor α
Publication TypeArticolo su Rivista peer-reviewed
Year of Publication2003
AuthorsHalin, C., Gafner V., Villani Maria Elena, Borsi L., Berndt A., Kosmehl H., Zardi L., and Neri D.
JournalCancer Research
Pagination3202 - 3210
Date Published2003
ISBN Number00085472 (ISSN)
Keywordsadolescent, animal cell, animal model, animal tissue, Animals, Antibodies, antineoplastic activity, Antitumor, article, cancer antibody, cancer cell culture, cancer immunotherapy, cancer model, cancer regression, Cultured, Drug Delivery Systems, drug distribution, drug effect, drug potentiation, Drug Screening Assays, Drug Synergism, drug targeting, experimental neoplasm, Female, fibronectin, human, human cell, hybrid protein, Immunoconjugates, immunoglobulin F(ab) fragment, immunoglobulin variable region, immunotherapy, in vitro study, Interferon Type II, interleukin 12, Interleukin-12, maximum tolerated dose, Mice, Monoclonal, Monoclonal antibody, monoclonal antibody L19, mouse, Neoplasm Transplantation, nonhuman, priority journal, Recombinant Fusion Proteins, T-Lymphocytes, teratocarcinoma, Tissue Distribution, toxicity testing, Tumor Cells, tumor growth, tumor necrosis factor alpha, Tumor Necrosis Factor-alpha, unclassified drug, vaccination

The potent antitumor activity of certain cytokines is often achieved at the expense of unacceptable toxicity. One avenue to improve the therapeutic index of cytokines in cancer therapy consists of fusing them to monoclonal antibodies capable of a selective localization at the tumor site. We have constructed fusion proteins of interleukin-12 (IL-12) and tumor necrosis factor (TNF-α) with L19, an antibody fragment specific to the extradomain B of fibronectin which has been shown to target tumors in animal models and in patients with cancer. These fusions display a potent antitumor activity in several immunocompetent murine models of cancer but do not lead to complete remissions of established aggressive tumors. In this article, we have evaluated the tumor-targeting properties and the anticancer activities of combinations of the two antibody-cytokine fusion proteins, as well as of a triple fusion protein between IL-12, L19, and TNF-α. Although all fusion proteins were active in vitro, the triple fusion protein failed to localize to tumors in vivo and to show significant therapeutic effects. By contrast, the combination of IL-12-L19 and L19-TNF-α displayed potent synergistic anticancer activity and led to the eradication of F9 teratocarcinomas grafted in immunocompetent mice. When cured mice were rechallenged with tumor cells, a delayed onset of tumor growth was observed, indicating the induction of a partial antitumor vaccination effect. Potent anticancer effects were achieved at doses of IL-12-L19 and L19-TNF-α (2 μg + 2 μg/mouse), which were at least 5-fold lower than the maximal-tolerated dose. The combined administration of the two fusion proteins showed only a modest increase in toxicity, compared with treatments performed with the individual fusion proteins. These results show that the targeted delivery of cytokines to the tumor environment strongly potentiates their antitumor activity and that the combination treatment with IL-12-L19 and L19-TNF-α appears to be synergistic in vivo.


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