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Losch FO, Muller R, Mutschler B, Neri D, Natali PG, Reth M, Carsetti R: Activation of T cells via tumor antigen specific chimeric receptors: the role of the intracellular signaling domain. Int J Cancer. 2003 Jan 20;103(3):399-407.
T cells engineered to express hybrid receptors with antibody defined specificity can successfully be targeted to tumor cells. In order to select intracellular domains of chimeric receptors capable of efficiently activate T cells in vitro and in vivo, we compared the function of receptors, which share the same extracellular antigen-binding part, joined to different intra-cellular signal transduction units. The antigen binding domain of the receptors was a single-chain fragment of a monoclonal antibody, which recognize a High Molecular Weight Melanoma-Associated Antigen with high affinity. The intracellular tails were derived from the T-cell receptor zeta chain (TCR-zeta), from the B-cell receptor Ig-alpha molecule and from a mutated Ig-alpha molecule able of stronger signal transduction. We compared the activity of the different chimeric receptors at a single-cell level by using a T-cell line that expressed an activation-dependent EGFP-reporter gene. Upon cross-linking with immobilized antibodies, all receptors were able to induce EGFP expression in the majority of the T cells. In contrast, EGFP expression was induced by contact to melanoma cells in vitro only in T cells that expressed the chimeric receptor that contained the TCR-zeta intracellular tail. In these T cells, the co-expression of chimeric receptors that contain a mutated Ig-alpha tail lowers the threshold of T-cell activation and facilitates tumor recognition in vitro and in vivo. Given their specificity and efficiency, T cells grafted with these type of receptors may represent potential candidates for cancer passive immunotherapy. |
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