Novel antibody-like single-chain TCR antibody Fc fusion protein

Abstract

Abstract We have previously reported the construction of a fusion protein composed of a soluble single-chain T cell receptor genetically linked to the constant domain of the human IgG1 heavy chain (TCR-Ig). The antigen recognition portion of the protein binds to an unmutated peptide derived from human p53 (amino acids 264–272) presented in the context of HLA-A2.1, whereas the IgG1 Fc provides effector functions. The protein is capable of forming dimers, specifically staining tumor cells, and promoting target and effector cell conjugation. The protein also has potent antitumor effects against p53+/HLA-A2.1+ human tumor xenografts in athymic nude mice and can mediate cell killing by antibody-dependent cellular cytotoxicity. Therefore, TCR-Ig behaves like an antibody, but possesses the ability to recognize antigens derived from intracellular targets. To test TCR-Ig in fully immunocompetent models, we have generated murine tumor cells stably expressing the human p53 epitope by constructing a single chain trimer composed of the human p53 peptide genetically linked to murine beta 2 microglobulin genetically linked to HLA-A2.1, with the inclusion of a disulfide trap to enhance stability. Utilizing haNK (NK-92 cells stably expressing high affinity Fc receptor and IL-2) as effector cells in the presence of TCR-Ig we demonstrated specific killing of murine single chain trimer expressing tumor cells. We are currently evaluating the antitumor activities and vaccinal effects of TCR-Ig in an HLA-A2.1 transgenic mouse model. TCR-Ig may represent a novel group of immunotherapeutics that has the potential to expand the range of tumors available for targeted therapies beyond those currently addressed by conventional antibody-based approaches.