title: Molecular design of the Calphabeta interface favors specific pairing of introduced TCRalphabeta in human T cells.
abstract: A promising approach to adoptive transfer therapy of tumors is to reprogram autologous T lymphocytes by TCR gene transfer of defined Ag specificity. An obstacle, however, is the undesired pairing of introduced TCRalpha- and TCRbeta-chains with the endogenous TCR chains. These events vary depending on the individual endogenous TCR and they not only may reduce the levels of cell surface-introduced TCR but also may generate hybrid TCR with unknown Ag specificities. We show that such hybrid heterodimers can be generated even by the pairing of human and mouse TCRalpha- and TCRbeta-chains. To overcome this hurdle, we have identified a pair of amino acid residues in the crystal structure of a TCR that lie at the interface of associated TCR Calpha and Cbeta domains and are related to each other by both a complementary steric interaction analogous to a "knob-into-hole" configuration and the electrostatic environment. We mutated the two residues so as to invert the sense of this interaction analogous to a charged "hole-into-knob" configuration. We show that this inversion in the CalphaCbeta interface promotes selective assembly of the introduced TCR while preserving its specificity and avidity for Ag ligand. Noteworthily, this TCR modification was equally efficient on both a Mu and a Hu TCR. Our data suggest that this approach is generally applicable to TCR independently of their Ag specificity and affinity, subset distribution, and species of origin. Thus, this strategy may optimize TCR gene transfer to efficiently and safely reprogram random T cells into tumor-reactive T cells.
authors: Medical Clinic (III) and Polyclinic, Department of Hematology and Oncology, Johannes Gutenberg-University, Mainz, Germany. hvoss@uni-mainz.de
authors: Ralf-Holger
authors: RH
authors: Voss
authors: Ralph A
authors: RA
authors: Willemsen
authors: Jürgen
authors: J
authors: Kuball
authors: Margarete
authors: M
authors: Grabowski
authors: Renate
authors: R
authors: Engel
authors: Ratna S
authors: RS
authors: Intan
authors: Philippe
authors: P
authors: Guillaume
authors: Pedro
authors: P
authors: Romero
authors: Christoph
authors: C
authors: Huber
authors: Matthias
authors: M
authors: Theobald
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D018360:Crystallography, X-Ray; D019281:Dimerization; D018014:Gene Transfer Techniques; D006801:Humans; D051379:Mice; D017354:Point Mutation; D011487:Protein Conformation; D021381:Protein Transport; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D016693:Receptors, Antigen, T-Cell, alpha-beta