Constructing a fusion molecular with T cell receptor for activation of carcinoembryonic antigen specific cytotoxic T lymphocytes to gastrointestinal tumors / 中华消化杂志

ABSTRACT

Objective To generate specific targeting T cells against gastrointestinal cancer, a eukaryotic expression vector encoding carcinoembryonic antigen(CEA) specific single chain variable fragment(scFv) fused to the transmembrane and intracellular domains of the signal transducing chain of CD3? was constructed. Methods Anti CEA scFv cDNA was obtained by RT PCR. The transmembrane and intracellular domains of CD3?cDNA was amplified from human T lymphocyte using RT PCR to clone into a eukaryotic expression vector, and anti CEA scFv cDNA was inserted upstream of CD3?. Finally, the eukaryotic expression vector containing scFv CD3? pcDNA3.0 was constructed and sequenced. Results A 810 base pair of anti CEA scFv was in accordance with sequence concerned, a 438 base pair of cDNA of the transmembrane and intracellular domains of CD3? was confirmed as sequence concerning of GenBank. Further identification was carried out through agarose electrophoresis after enzyme digestion. Conclusion We constructed a fusion molecular with both CEA targeting and CD3? signal transduction perporties, which lay a good foundation for generation of modified cytotoxic T lymphocytes against gastrointestinal tumors.