HIGH-EFFICIENCY EXPRESSION AND SOLUBILIZATION OF FUNCTIONAL T-CELL ANTIGEN RECEPTOR HETERODIMERS

The T cell receptor (TCR) zeta-chain was attached to the TCR-alpha and beta-extracellular domains to induce efficient expression of alpha-beta-heterodimers that can recognize complexes of antigen with major histocompatibility complex (MHC) molecules. Chimeric constructs expressed in RBL-2H3 cells were efficiently transported to the cell surface uniquely as disulfide-linked heterodimers. Transfectants were activated by specific antigen-MHC complexes, which demonstrated that the expressed-alpha-beta was functional and that CD3 was not required for antigen-MHC binding. Constructs with thrombin cleavage sites were efficiently cleaved to soluble disulfide-linked heterodimers. Thus, attachment of TCR-zeta-domains and protease cleavage sites to TCR-alpha and beta induces expression of demonstrably functional heterodimers that can be solubilized.