A novel strategy for the generation of T cell lines lacking expression of endogenous α- and/or β-chain T cell receptor genes

Mutant T cell lines that do not express the endogenous α- and/or β-chain genes of the TCR were generated from the αβ TCR/CD3⁺ tumor cell line C6VL with a combination of classical mutagenesis methods and selection of somatic hybrid variants. This novel strategy obviated the need for repeated mutagenesis and screening of a large number of individual clones. The loss of either the α- or the β-chain expression in the mutant cells was associated with the loss of surface TCR/CD3 complex, which could be rescued by the transaction of appropriate exogenous α- and/or β-chain gene constructs. Because these cells express a single TCR molecule on the cell surface, they are useful for the study of the assembly and function of the αβ TCR. This strategy is also generally applicable for the generation of homozygous mutant cell lines lacking other gene products.