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  • Cell-Based Therapy
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Cell-Based Therapy

Engineering mouse T lymphocytes specific to type II collagen by transduction with a chimeric receptor consisting of a single chain Fv and TCR zeta

Gene Therapy volume 7, pages 714–722 (2000)Cite this article

Abstract

The chimeric cell surface receptor scC2Fv/CD8/ζ was constructed to engineer primary mouse T lymphocytes with antibody-type specificity to type II collagen (CII). Such cells could be used as gene carriers in the anti-inflammatory gene therapy of an autoimmune arthritis. This receptor includes the single chain Fv domain (scFv) of the anti-CII monoclonal antibody (mAb) C2, hinge region of CD8α and the transmembrane and cytoplasmic domains of TCRζ. The scC2Fv/CD8/ζ gene was transduced into T cell hybridomas and primary mouse lymphocytes using retrovirus-mediated gene transfer. The chimeric receptor scC2Fv/CD8/ζ forms covalently bound homodimers, as demonstrated in T cell hybridomas and packaging fibroblasts. It does not associate with endogenous signalling subunits of the TCR complex. When scC2Fv/CD8/ζ-expressing clones of T cell hybridomas MD.45 and HCQ6 were stimulated with CII they produced IL-2. The level of their IL-2 response correlated with the expression level of the chimeric receptor on the cell surface. Splenocytes isolated from DBA/1 mice were stimulated with Con A in vitro to facilitate retrovirus-mediated transfer of the scC2Fv/CD8/ζ gene. As a result of transduction, approximately 4% of the Con A-activated splenocytes expressed the chimeric receptor scC2Fv/CD8/ζ on the cell surface. These cells proliferated in response to stimulation with CII.

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Acknowledgements

We thank Drs Thomas Broker for donating the plasmid p2δRICD8-ζ and Marco Londei for critically reading the manuscript. This work was supported by the Arthritis Research Campaign and Multiple Sclerosis Society, United Kingdom.

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  1. Kennedy Institute of Rheumatology, London, UK

    A Annenkov & Y Chernajovsky

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  1. A Annenkov
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  2. Y Chernajovsky
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Annenkov, A., Chernajovsky, Y. Engineering mouse T lymphocytes specific to type II collagen by transduction with a chimeric receptor consisting of a single chain Fv and TCR zeta. Gene Ther 7, 714–722 (2000). https://doi.org/10.1038/sj.gt.3301149

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