Intracellular single chain Fv antibody inhibits Ras activity in T-cell antigen receptor stimulated Jurkat cells.
A mammalian expression vector directing the synthesis of a cytoplasmic single chain Fv version of the Y13-259 anti-Ras antibody was constructed and co-transfected into the human lymphoid cell line Jurkat together with a reporter construct containing the bacterial gene for chloramphenicol acetyl transferase under the transcriptional control of several copies of the binding site for the transcription factor NF-AT. The Ras specific antibody interferes with NF-AT activation upon direct activation of the T-cell antigen receptor, whereas activation by direct protein kinase C stimulation is less sensitive to the anti-Ras antibody. Furthermore, the observed inhibition is dependent on the ratio of antibody to reporter plasmid utilized in the transfection experiments.
Werge
T M
TM
CNR Institute of Neurobiology, Rome, Italy.
Baldari
C T
CT
Telford
J L
JL
D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology*; D008213:Lymphocyte Activation; D015689:Oncogene Protein p21(ras) / Q000037:antagonists & inhibitors* / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism*
D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011948:Receptors, Antigen, T-Cell; C055966:immunoglobulin Fv; D015500:Chloramphenicol O-Acetyltransferase; D015689:Oncogene Protein p21(ras)