LacZ inducible, antigen/MHC-specific T cell hybrids

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摘要：

The nuclear factor of activated T cells (NFAT) enhancer element of the gene can regulate expression of the reporter gene in activated T cells. Based upon this observation, we showed that this inducible NFAT--construct could be used to measure mediated, ligand-specific activation in single T cells. Here we describe a general approach to obtaining inducible, T cell hybrids by generating two new fusion partners BWZ.36 and BWZ.36 CD8 alpha derived by transfecting alpha-beta-BW5147 cells with the NFAT--construct. Using these fusion partners and normal T cells from immunized , we obtained T cell hybrids in which activity is specifically induced in response to antigen/MHC class II or MHC class I complexes. We show that measuring ligand induced by the non-radioactive assay is simpler, faster, and most cost-effective relative to conventional assays. Most importantly, the unique ability to detect activation of single T cells by the assays permits detection of rare antigen presenting cells and thus provides the basis for developing expression cloning strategies for identifying unknown T cell antigens.

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关键词：

expression cloning β-galactosidase T cell activation

DOI：

10.1093/intimm/6.3.369

被引量：

650

年份：

1994