Rhadinovirus vector-derived human telomerase reverse transcriptase expression in primary T cells.
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.
Toptan
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T
Institute for Infection Medicine, Christian-Albrecht University of Kiel, Kiel, Germany.
Ensser
A
A
Fickenscher
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H
D018801:CD2 Antigens / Q000276:immunology; D002472:Cell Transformation, Viral / Q000235:genetics*; D005822:Genetic Vectors; D006567:Herpesvirus 2, Saimiriine / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007378:Interleukins / Q000276:immunology; D010750:Phosphoproteins / Q000235:genetics; D013601:T-Lymphocytes / Q000201:enzymology*; D019098:Telomerase / Q000235:genetics*; D014161:Transduction, Genetic / Q000379:methods*; D014764:Viral Proteins / Q000235:genetics; D019061:src-Family Kinases / Q000032:analysis / Q000276:immunology
D018801:CD2 Antigens; C406767:IL26 protein, human; D007378:Interleukins; D010750:Phosphoproteins; D014764:Viral Proteins; C092384:tyrosine kinase interacting protein, Saimiriine herpesvirus 2; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases; D019098:Telomerase