Preparation of HLA-unrestricted anti-tumor cytotoxic T cells by gene transfer.

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In this study, we attempted the establishment of the stable T cell clones transfected with a fusion gene consisting of cDNAs coding for single chain antibody and T cell receptor gannma chain. The VH and VL cDNAs of an anti-MUC1 mucin monoclonal antibody MUSE11 were amplified from total RNA extracted from hybridoma by RT-PCR method, then ligated them and used as the single chain antibody gene. The fusion gene was ligated into a retroviral vector PG1EN and transient transfection into packaging cells was performed to obtein viruses. However, the virus titer sufficient for establishing stable transfectans of EB virus-transformed T cells has thus far been obtained, being now evaluating various conditions of virus production. On the other hand, we have recently obtained thr stable transfectants using eukaryocytic expression vectoe haboring the fusin gene by conventional G418 selection. Some transfectant clones were shown to express the binding activity to MUC1-positive cultured tumor cells. Detailed studies on the expression level of signal chain antibody, the specificity of the binding of transfectants to cultured cells and so on are now under ongoing. Signal transduction of T cell receptor gamma chain and the binding ability of transfectants in vivo will be aiso examined.