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Targeting the Wilms Tumor Antigen 1 by TCR Gene Transfer: TCR Variants Improve Tetramer Binding but Not the Function of Gene Modified Human T Cells¹

Sharyn Thomas^*, Shao-An Xue^*, Michela Cesco-Gaspere^*, Esther San José, Daniel P. Hart^*, Vivian Wong^2,*, Reno Debets, Balbino Alarcon, Emma Morris^* and Hans J. Stauss^3,*

^* Department of Immunology and Molecular Pathology, University College London, Hampstead Campus, Royal Free Hospital, London; Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Madrid, Spain; and Tumor Immunology Group, Department of Medical Oncology, Eramus Medical Center-Daniel den Hoed Cancer Center, Rotterdam, The Netherlands

We have previously described the functional activity of a human TCR specific for an HLA-A2-presented peptide derived from the Wilms tumor Ag 1 (WT1). Recent studies showed that the expression and function of human TCR was improved by the introduction of an additional disulfide bond between the - and -chains or by the exchange of the human constant region for murine sequences. In this study, we analyzed the functional activity of WT1-TCR variants expressed in Jurkat cells and in primary T cells. The introduction of cysteine residues or murine constant sequences into the WT1-TCR did not result in a global reduction of mispairing with wild-type TCR chains. Instead, the level of mispairing was affected by the variable region sequences of the wild-type TCR chains. The analysis of freshly transduced peripheral blood T cells showed that the transfer of modified TCR constructs generated a higher frequency of Ag-responsive T cells than the transfer of the wild-type TCR. After several rounds of peptide stimulation this difference was no longer observed, as all transduced T cell populations accumulated 90% of Ag-responsive T cells. Although the Ag-responsive T cells expressing the modified TCR bound the HLA-A2/WT1 tetramer more efficiently than T cells expressing the wild-type TCR, this did not improve the avidity of transduced T cells nor did it result in a measurable enhancement in IFN- production and cytotoxic activity. This indicated that the enhanced tetramer binding of modified WT1-TCR variants was not associated with improved WT1-specific T cell function.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported by the European Union-funded ATTACK project and by the Leukemia Research Fund.

² Current address: Laboratory of Functional Immunogenetics, The Babraham Institute, Babraham Research Campus, Cambridge, U.K.

³ Address correspondence and reprint requests to Prof. Hans J. Stauss. Department of Immunology and Molecular Pathology, University College London, Hampstead Campus, Royal Free Hospital, Rowland Hill Street, London, United Kingdom. E-mail address: h.stauss{at}medsch.ucl.ac.uk

⁴ Abbreviations used in this paper: WT1, Wilms tumor Ag 1; MFI, mean fluorescence intensity.

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