NF-AT-luciferase reporter T cell lines as tools to screen immunosuppressive drugs.
The development of safer analogues of immunosuppressants such as cyclosporin A and FK506 is an important goal for a number of clinical applications ranging from transplantation to the treatment of autoimmune diseases. Here we show the generation and the characterization of Jurkat T cell lines stably transfected with a reporter construct containing the firefly luciferase gene under the control of NF-AT. These lines specifically respond in a cyclosporin A-sensitive manner to T cell antigen receptor-derived signals. Due to the high levels of luciferase activity expression fewer than 1000 cells are required for detection of luciferase. In addition, a simplified luciferase assay allows to reduce both the manipulations and the time required for the assay, making these lines potentially useful models for the automated screening of cyclosporin A and FK506 analogues.
Baldari
C T
CT
IRIS-Biocine, Siena, Italy.
Di Somma
M M
MM
Majolini
M B
MB
Ulivieri
C
C
Milia
E
E
Telford
J L
JL
D016572:Cyclosporine / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D017930:Genes, Reporter; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D019169:Jurkat Cells; D008156:Luciferases / Q000235:genetics*; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D013601:T-Lymphocytes; D016559:Tacrolimus / Q000031:analogs & derivatives; D014157:Transcription Factors / Q000378:metabolism*; D014162:Transfection
D004268:DNA-Binding Proteins; D007166:Immunosuppressive Agents; D050778:NFATC Transcription Factors; D009687:Nuclear Proteins; D014157:Transcription Factors; D016572:Cyclosporine; D008156:Luciferases; D016559:Tacrolimus