title: Human TCR that incorporate CD3zeta induce highly preferred pairing between TCRalpha and beta chains following gene transfer.
abstract: TCR gene therapy is adversely affected by newly formed TCRalphabeta heterodimers comprising exogenous and endogenous TCR chains that dilute expression of transgenic TCRalphabeta dimers and are potentially self-reactive. We have addressed TCR mispairing by using a modified two-chain TCR that encompasses total human CD3zeta with specificities for three different Ags. Transfer of either TCRalpha:CD3zeta or beta:CD3zeta genes alone does not result in surface expression, whereas transfer of both modified TCR chains results in high surface expression, binding of peptide-MHC complexes and Ag-specific T cell functions. Genetic introduction of TCRalphabeta:zeta does not compromise surface expression and functions of an endogenous TCRalphabeta. Flow cytometry fluorescence resonance energy transfer and biochemical analyses demonstrate that TCRalphabeta:CD3zeta is the first strategy that results in highly preferred pairing between CD3zeta-modified TCRalpha and beta chains as well as absence of TCR mispairing between TCR:CD3zeta and nonmodified TCR chains. Intracellular assembly and surface expression of TCR:CD3zeta chains is independent of endogenous CD3gamma, delta, and epsilon. Taken together, our data support the use of TCRalphabeta:CD3zeta to prevent TCR mispairing, which may provide an adequate strategy to enhance efficacy and safety of TCR gene transfer.
authors.affiliation: Unit Clinical and Tumor Immunology, Department of Medical Oncology, Erasmus Medical Center-Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Sebestyén
authors.forename: Erik
authors.initials: E
authors.lastname: Schooten
authors.forename: Tamara
authors.initials: T
authors.lastname: Sals
authors.forename: Irene
authors.initials: I
authors.lastname: Zaldivar
authors.forename: Esther
authors.initials: E
authors.lastname: San José
authors.forename: Balbino
authors.initials: B
authors.lastname: Alarcón
authors.forename: Sara
authors.initials: S
authors.lastname: Bobisse
authors.forename: Antonio
authors.initials: A
authors.lastname: Rosato
authors.forename: János
authors.initials: J
authors.lastname: Szöllosi
authors.forename: Jan Willem
authors.initials: JW
authors.lastname: Gratama
authors.forename: Ralph A
authors.initials: RA
authors.lastname: Willemsen
authors.forename: Reno
authors.initials: R
authors.lastname: Debets
mesh_terms: D017252:CD3 Complex / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D018014:Gene Transfer Techniques; D006801:Humans; D019169:Jurkat Cells; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
chemical_list: D017252:CD3 Complex; C121704:CD3 antigen, zeta chain; D016693:Receptors, Antigen, T-Cell, alpha-beta