Human TCR that incorporate CD3ζ induce highly preferred pairing between TCRα and β chains following gene transfer

Zsolt Sebestyén, Erik Schooten, Tamara Sals, Irene Zaldivar, Esther San José, Balbino Alarcón, Sara Bobisse, Antonio Rosato, J. Szöllősi, Jan Willem Gratama, Ralph A. Willemsen, Reno Debets

Research output: Contribution to journalArticle

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Abstract

TCR gene therapy is adversely affected by newly formed TCRαβ heterodimers comprising exogenous and endogenous TCR chains that dilute expression of transgenic TCRαβ dimers and are potentially self-reactive. We have addressed TCR mispairing by using a modified two-chain TCR that encompasses total human CD3ζ with specificities for three different Ags. Transfer of either TCRα:CD3ζ or β:CD3ζ 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 TCRαβ: ζ does not compromise surface expression and functions of an endogenous TCRαβ. Flow cytometry fluorescence resonance energy transfer and biochemical analyses demonstrate that TCRαβ:CD3ζ is the first strategy that results in highly preferred pairing between CD3ζ-modified TCRα and β chains as well as absence of TCR mispairing between TCR:CD3ζ and nonmodified TCR chains. Intracellular assembly and surface expression of TCR:CD3ζ chains is independent of endogenous CD3γ, δ, and ε. Taken together, our data support the use of TCRαβ:CD3ζ to prevent TCR mispairing, which may provide an adequate strategy to enhance efficacy and safety of TCR gene transfer.

Original languageEnglish
Pages (from-to)7736-7746
Number of pages11
JournalJournal of Immunology
Volume180
Issue number11
Publication statusPublished - 2008

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Fluorescence Resonance Energy Transfer
Genetic Therapy
Genes
Flow Cytometry
T-Lymphocytes
Safety
MHC binding peptide

ASJC Scopus subject areas

  • Immunology

Cite this

Sebestyén, Z., Schooten, E., Sals, T., Zaldivar, I., José, E. S., Alarcón, B., ... Debets, R. (2008). Human TCR that incorporate CD3ζ induce highly preferred pairing between TCRα and β chains following gene transfer. Journal of Immunology, 180(11), 7736-7746.

Human TCR that incorporate CD3ζ induce highly preferred pairing between TCRα and β chains following gene transfer. / Sebestyén, Zsolt; Schooten, Erik; Sals, Tamara; Zaldivar, Irene; José, Esther San; Alarcón, Balbino; Bobisse, Sara; Rosato, Antonio; Szöllősi, J.; Gratama, Jan Willem; Willemsen, Ralph A.; Debets, Reno.

In: Journal of Immunology, Vol. 180, No. 11, 2008, p. 7736-7746.

Research output: Contribution to journalArticle

Sebestyén, Z, Schooten, E, Sals, T, Zaldivar, I, José, ES, Alarcón, B, Bobisse, S, Rosato, A, Szöllősi, J, Gratama, JW, Willemsen, RA & Debets, R 2008, 'Human TCR that incorporate CD3ζ induce highly preferred pairing between TCRα and β chains following gene transfer', Journal of Immunology, vol. 180, no. 11, pp. 7736-7746.
Sebestyén Z, Schooten E, Sals T, Zaldivar I, José ES, Alarcón B et al. Human TCR that incorporate CD3ζ induce highly preferred pairing between TCRα and β chains following gene transfer. Journal of Immunology. 2008;180(11):7736-7746.
Sebestyén, Zsolt ; Schooten, Erik ; Sals, Tamara ; Zaldivar, Irene ; José, Esther San ; Alarcón, Balbino ; Bobisse, Sara ; Rosato, Antonio ; Szöllősi, J. ; Gratama, Jan Willem ; Willemsen, Ralph A. ; Debets, Reno. / Human TCR that incorporate CD3ζ induce highly preferred pairing between TCRα and β chains following gene transfer. In: Journal of Immunology. 2008 ; Vol. 180, No. 11. pp. 7736-7746.
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