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A soluble single-chain T-cell receptor IL-2 fusion protein retains MHC-restricted peptide specificity and IL-2 bioactivity

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Abstract

Antibody-based targeted immunotherapy has shown promise as an approach to treat cancer. However, many known tumor-associated antigens are not expressed as integral membrane proteins and cannot be utilized as targets for antibody-based therapeutics. In order to expand the limited target range of antibodies, we have constructed a soluble single-chain T-cell receptor (TCR) fusion protein designated 264scTCR/IL-2. This fusion protein is comprised of a three-domain HLA-A2-restricted TCR specific for a peptide epitope of the human p53 tumor suppressor protein, which is overexpressed in a broad range of human malignancies. The 264scTCR/IL-2 fusion protein has been expressed at high levels in mammalian cells, and milligram quantities have been purified. MHC-restricted antigen-specific binding properties are maintained in the single-chain, three-domain TCR portion of the fusion protein, and the IL-2 portion retains bioactivity similar to that of free recombinant IL-2. Moreover, this fusion protein is capable of conjugating target and effector cells, remains intact in the blood and substantially increases the half life of the IL-2 portion of the molecule. Finally, the 264scTCR/IL-2 fusion protein can be used to stain tumor cells and is capable of reducing lung metastases in an experimental model of metastasis. Thus, TCR-based fusion proteins may provide a novel class of targeted immunotherapeutics for cancer.

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Acknowledgements

This work was supported in part by grant no. 1R43CA88615-01 from the National Institutes of Health.

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Authors and Affiliations

  1. Altor BioScience Corporation, 2810 N. Commerce Parkway, Miramar, FL 33025, USA

    Kimberlyn F. Card, Shari A. Price-Schiavi, Bai Liu, Elizabeth Thomson, Heather Belmont, Janette Builes & Hing C. Wong

  2. Dade-Behring Corporation, Newark, DE 19702, USA

    Esperanza Nieves

  3. The Scripps Research Institute, La Jolla, CA 92037, USA

    Javier Hernandez & Linda Sherman

  4. Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University, Amarillo, TX 79106, USA

    Jon Weidanz

  5. Florida Hospital Cancer Institute, Orlando, FL 32804, USA

    John L. Francis & Ali Amirkhosravi

  6. Hematech LLC, Sioux Falls, SD 57106, USA

    Jin-an Jiao

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  1. Kimberlyn F. Card
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Correspondence to Hing C. Wong.

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Card, K.F., Price-Schiavi, S.A., Liu, B. et al. A soluble single-chain T-cell receptor IL-2 fusion protein retains MHC-restricted peptide specificity and IL-2 bioactivity. Cancer Immunol Immunother 53, 345–357 (2004). https://doi.org/10.1007/s00262-003-0450-3

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