The overall goal of this project is to demonstrate proof-of-principle for using T-cell receptor (TCR)-based therapeutics in the treatment of cancer. To achieve this goal, they cloned a single chain TCR (scTCR) from a high avidity cytotoxic T lymphocyte (CTL) cell line derived from an HLA-A2 transgenic mouse. The TCR recognizes a naturally processed peptide fragment from human wild-type p53 in the context of HLA-A2.1. Recently, they expressed the TCR as a soluble single-chain TCR-kappa fusion protein in Chinese hamster ovary (CHO) cells. The purified protein has been characterized and its affinity constant (Kd=107 M-1) has been determined. Due to its high affinity (especially its longer dissociation half-life), they proceeded to use this scTCR-kappa fusion protein for preparation of an immunoconjugate. The process involved cross-linking doxorubicin (Dox) molecules onto the scTCR-kappa fusion protein using a proprietary coupling technology. To evaluate the efficacy of tumor suppression by this novel immunoconjugate in vitro, they will carry out cytotoxic assays using peptide-pulsed cells, and later, human breast carcinoma cell lines. In addition, studies are proposed to measure the anti-tumor activity of the TCR-Dox immunoconjugate using two different murine systems. These models are proposed to provide valuable information on the dosage and route of administration for the TCR-Dox conjugate and generate preliminary in vivo data to support proof-of-principle for using scTCR-conjugates in the treatment of human cancer.
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| Mosquera, Luis A; Card, Kimberlyn F; Price-Schiavi, Shari A et al. (2005) In vitro and in vivo characterization of a novel antibody-like single-chain TCR human IgG1 fusion protein. J Immunol 174:4381-8 |
| Card, Kimberlyn F; Price-Schiavi, Shari A; Liu, Bai et al. (2004) A soluble single-chain T-cell receptor IL-2 fusion protein retains MHC-restricted peptide specificity and IL-2 bioactivity. Cancer Immunol Immunother 53:345-57 |
