Intracellular single chain Fv antibody inhibits Ras activity in T‐cell antigen receptor stimulated Jurkat cells

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Abstract
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.