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Rhadinovirus vector-derived human telomerase reverse transcriptase expression in primary T cells

Gene Therapy volume 17, pages 653–661 (2010)Cite this article

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Abstract

The rhadinovirus herpesvirus saimiri (HVS) as a gene delivery vector allows large DNA insertions and long-termed gene expression. In the case of T-cell transduction, such vectors use the viral transformation-associated genes of HVS C488 for T-cell amplification. In this report, we investigated whether the gene for the catalytic telomerase subunit human telomerase reverse transcriptase (hTERT) can substitute for the transformation-associated genes in rhadinoviral T-cell transduction and amplification. By using virus mutants generated by en passant mutagenesis from bacterial artificial chromosomes, we observed a very early and functional transgene expression even by virus mutants without transformation-associated genes. The markers of T-cell transformation by HVS, namely CD2 hyperreactivity, overexpression of interleukin-26, and of the tyrosine kinase Lyn could neither be induced nor enhanced by ectopic hTERT expression. When the viral transformation-associated genes were replaced by the hTERT gene, it was not sufficient for growth transformation, although hTERT was efficiently transduced and functionally expressed by the rhadinovirus vector. Thus, the transformation-associated proteins StpC and Tip are responsible for the T-cell phenotype after transduction by HVS and, additionally, modulate telomerase activity independently of hTERT expression.

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Acknowledgements

This study was supported by a grant to HF from Wilhelm Sander-Stiftung, Neustadt/Donau, Germany, by an initial fellowship to TT by the German Academic Exchange Service (DAAD), Bonn, Germany and, in part, by the Research Center Grant SFB 617 A24 to HF, and the Excellence Center Inflammation at Interfaces (Kiel). In addition, AE was supported, in part, by DFG EN423-2/1 and Research Center Grant SFB 796 and a grant by the Wilhelm Sander-Stiftung. We thank Dr Marco Migliaccio (Lausanne) for providing a functional hTERT construct, Dr Mirko Kummer (Erlangen) for cloning the PGK-driven hTERT expression cassette, Dr Karsten Tischer (Berlin) for providing the reagents needed for en passant mutagenesis and Dr Pierre van der Bruggen (Ludwig Institute of Cancer Research, Brussels) for stimulating discussions.

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Authors and Affiliations

  1. Institute for Infection Medicine, Christian-Albrecht University of Kiel, Kiel, Germany

    T Toptan & H Fickenscher

  2. Department of Infectious Diseases, Virology, Ruprecht-Karl University of Heidelberg, Heidelberg, Germany

    T Toptan & H Fickenscher

  3. Institute for Clinical and Molecular Virology, Friedrich-Alexander University of Erlangen-Nuremberg, Erlangen, Germany

    A Ensser

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  1. T Toptan
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Correspondence to H Fickenscher.

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Toptan, T., Ensser, A. & Fickenscher, H. Rhadinovirus vector-derived human telomerase reverse transcriptase expression in primary T cells. Gene Ther 17, 653–661 (2010). https://doi.org/10.1038/gt.2010.3

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