Institute for Infection Medicine, Christian-Albrecht University of Kiel, Kiel, Germany.
Gene Ther. 2010 May;17(5):653-61. doi: 10.1038/gt.2010.3. Epub 2010 Feb 18.
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.
猴疱疹病毒 1 型（HVS）作为一种基因传递载体，可以允许较大的 DNA 插入和长期的基因表达。在 T 细胞转导的情况下，此类载体使用 HVS C488 的病毒转化相关基因进行 T 细胞扩增。在本报告中，我们研究了人端粒酶逆转录酶（hTERT）的催化端粒酶亚基基因是否可以替代猴疱疹病毒 T 细胞转导和扩增中的转化相关基因。通过使用细菌人工染色体进行偶然诱变产生的病毒突变体，我们观察到即使是没有转化相关基因的病毒突变体也能实现非常早期和功能性的转基因表达。HVS 引起的 T 细胞转化的标志物，即 CD2 超反应性、白细胞介素-26 的过表达和酪氨酸激酶 Lyn 的过表达，既不能被诱导，也不能被 hTERT 表达增强。当病毒转化相关基因被 hTERT 基因取代时，即使 hTERT 被猴疱疹病毒载体有效地转导和功能性表达，也不足以实现生长转化。因此，StpC 和 Tip 等转化相关蛋白负责 HVS 转导后的 T 细胞表型，并且独立于 hTERT 表达调节端粒酶活性。
