Application of Epstein Barr Virus (EBV) for preparation and characterization of cells with ability to express telomerase (hTERT) mRNA.
2007
Aqa`i Pur, Khosru | Shokr Gozar, Mohammad Ali | Safaviyeh, Sediqeh Sadat | Kargar Mo`akhhar, Ruhani
Epstein - Barr virus is one of the most important viruses that could transform some of the cells. The virus infects B Lymphocytes through CD21 surface receptor and immortalize human B Lymphocytes via transformation and makes them B Lymphoblastoid cell lines (BLCLs). This study has been investigated the mechanism of immortalization of B Lymphocytes transformed by EBV and its relation with switching on their telomerase gene. Telomerase is a ribonucleoprotein acts as reveres transcriptase, and adds telomeric repeat at the end of eukaryotic chromosomes to maintain the prolifrative capacity of cells by preventing cells replicative senescenses. In this study, the peripheral blood mononuclear cells (PBMC) were transformed by EBV and then the cell content (telomerase) was extracted. In the second part of the study, telomerase activity was measured using TRAP assay (Telomeric Repeat Amplification Protocol) techniqu and the retative telomerase activity of these transformed cell lines investigated altogether and with none transformed cells. Twenty two of the twenty four transformed cells were telomerase positive with strong telomerase activity. Four none transformed PBMC used, which all of them were telomerase negative or with very trace activity. Results are consistent with the hypothesis that the mechanism of immortalization of B Lymphocytes transformed by EBV is majority by telomerase gene switching on. Thus, telomerase activity could be discussed as a marker for their immortality. Following this study, it could be proposed that EBV is one of the main candidates for diploid cell immortalization, via telomerase switching on. These immortalized cell lines could be used as a source of telomerase positive cells and for molecular studies on telomerase and its subunits.
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