Expression of T-cell receptor alpha-chain genes in transgenic mice.

作者：Leslie J. Berg, Barbara Fazekas de St Groth, Fredrik Ivars, Christopher Carl Goodnow, Susan Gilfillan, Henri‐Jean Garchon, Mark M. Davis · 发表于：Molecular and Cellular Biology · 年份：1988 · DOI：10.1128/mcb.8.12.5459 · 被引用次数：60 · 研究领域：CAR-T cell therapy research、Monoclonal and Polyclonal Antibodies Research、Transgenic Plants and Applications

To examine the influences responsible for shaping the T-cell repertoire in vivo, we have introduced T-cell receptors of defined specificity into mice. In this report, we analyze transgenic mice carrying a T-cell receptor alpha-chain gene from a pigeon cytochrome c-reactive T-cell line. A variant of this construct, which has the immunoglobulin heavy-chain enhancer inserted into the JC intron, was also introduced into mice. Addition of the enhancer increased the steady-state level of transgene-encoded mRNA three- to fivefold in cultured T cells, leading to a two- to threefold increase in surface expression. In vivo, the difference between these two constructs was even more significant, increasing the number of transgene-positive cells from approximately 5 to 70% and the T-cell receptor surface density two- to threefold. Surprisingly, while surface expression of either type of transgene was limited to T cells, we found little tissue specificity with respect to transcription. In T cells expressing the alpha chain from the enhancer-containing construct, immunoprecipitation with a 2B4 alpha-specific monoclonal antibody revealed the expected disulfide-linked dimer. Costaining of these T cells with the 2B4 alpha-specific monoclonal antibody versus anti-CD3 indicated that expression of the transgene-encoded alpha chain precludes expression of endogenous alpha chains on the majority of cells; in contrast, 2B4 alpha-chain expression from the construct lacking the enhancer is inefficient...