title: TCR transfer induces TCR-mediated tonic inhibition of RAG genes in human T cells.
abstract: Induction of the TCR signaling pathway terminates the expression of RAG genes, and a link between this pathway and their transcriptional control is evident from the recent demonstration of their re-expression if the TCR is subsequently lost or down-regulated. Since unstimulated T cells display a steady-state level of "tonic" TCR signaling, i.e. in the absence of any antigenic stimulus, it was uncertain whether this control was exerted through ligand-dependent or ligand-independent TCR signaling. Here we demonstrate for the first time that exogenous TCR α and β chains transferred into the human immature RAG(+) T cell line Sup-T1 by lentiviral transduction inhibit RAG expression through tonic signaling, and that this inhibition could itself be reverted by pharmacological tonic pathway inhibitors. We also suggest that mature T cells already expressing an endogenous TCR on their surface maintain some levels of plasticity at the RAG locus when their basal TCR signaling is interfered with. Lastly, we show that the TCR constructs employed in TCR gene therapy do not possess the same basal signaling transduction capability, a feature that may have therapeutic implications.
authors.affiliation: Department of Clinical and Biological Sciences, University of Turin, Orbassano, Italy. luca.orlando@unito.it
authors.forename: Luca
authors.initials: L
authors.lastname: Orlando
authors.forename: Lisa
authors.initials: L
authors.lastname: Accomasso
authors.forename: Paola
authors.initials: P
authors.lastname: Circosta
authors.forename: Valentina
authors.initials: V
authors.lastname: Turinetto
authors.forename: Erica
authors.initials: E
authors.lastname: Lantelme
authors.forename: Paola
authors.initials: P
authors.lastname: Porcedda
authors.forename: Valentina
authors.initials: V
authors.lastname: Minieri
authors.forename: Marisa
authors.initials: M
authors.lastname: Pautasso
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Willemsen
authors.forename: Alessandro
authors.initials: A
authors.lastname: Cignetti
authors.forename: Claudia
authors.initials: C
authors.lastname: Giachino
mesh_terms: D001549:Benzamides; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D002648:Child; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D015536:Down-Regulation; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D016654:Genes, RAG-1; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D000068877:Imatinib Mesylate; D015151:Immunoblotting; D019169:Jurkat Cells; D008024:Ligands; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D010879:Piperazines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D016559:Tacrolimus / Q000494:pharmacology; D014161:Transduction, Genetic
chemical_list: D001549:Benzamides; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D008024:Ligands; D009687:Nuclear Proteins; D010879:Piperazines; D011743:Pyrimidines; C499311:RAG2 protein, human; D016693:Receptors, Antigen, T-Cell, alpha-beta; C064658:RAG-1 protein; D000068877:Imatinib Mesylate; D016559:Tacrolimus; D004121:Dimethyl Sulfoxide