Activation of T cells via tumor antigen specific chimeric receptors: The role of the intracellular signaling domain

• Published in: International journal of cancer 2003-01, Vol.103 (3), p.399-407
• Main Authors: Losch, Florian O., Müller, Ralph, Mutschler, Bettina, Neri, Dario, Natali, Pier Giorgio, Reth, Michael, Carsetti, Rita
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Neoplasm - immunology; Antibody Specificity; Antigens, Neoplasm - immunology; Antigens, Neoplasm - metabolism; Antineoplastic agents; Ascitic Fluid - immunology; Biological and medical sciences; CD3 Complex - immunology; Green Fluorescent Proteins; High Molecular Weight Melanoma‐Associated Antigen; Humans; Immunoglobulin Variable Region - immunology; Immunotherapy; In Vitro Techniques; Luminescent Proteins - metabolism; Lymphocyte Activation; Medical sciences; Melanoma - immunology; Melanoma - therapy; Melanoma, Experimental - immunology; Melanoma, Experimental - therapy; Mice; Mice, SCID; Microscopy, Confocal; monoclonal antibody; Pharmacology. Drug treatments; Receptors, Antigen, B-Cell - immunology; Receptors, Antigen, T-Cell, gamma-delta - immunology; Recombinant Fusion Proteins - immunology; Signal Transduction - physiology; single chain fragment variable; T-Lymphocytes - immunology; Tumor Cells, Cultured
• ISSN: 0020-7136; 1097-0215
• DOI: 10.1002/ijc.10826

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 ζ chain (TCR‐ζ), from the B‐cell receptor Ig‐α molecule and from a mutated Ig‐α 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‐ζ intracellular tail. In these T cells, the co‐expression of chimeric receptors that contain a mutated Ig‐α 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. © 2002 Wiley‐Liss, Inc.