Detection and organ distribution of herpes simplex virus thymidine kinase gene-transduced T cells to monitor alloreactivity after bone marrow transplantation in rats
| Autor: | Kolen, S.M.H., Weijtens, M., Spronsen, A. van, Smulders, S., Weger, R. de, Witte, T.J.M. de, Dolstra, H., Hagenbeek, A., Wiel-van Kemenade, E. van de, Martens, A.C.M. |
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| Rok vydání: | 2003 |
| Předmět: | |
| Zdroj: | Human Gene Therapy, 14, 4, pp. 341-51 Human Gene Therapy, 14, 341-51 |
| ISSN: | 1043-0342 |
| Popis: | Item does not contain fulltext Introduction of the HSV-Tk suicide gene into allogeneic T cells offers the possibility to control developing host-reactive cells within the context of allogeneic bone marrow transplantation (BMT). Sensitive quantitative detection methods are a prerequisite to monitor genetically modified T cells in peripheral blood and tissues to study their involvement in graft-versus-host disease (GVHD)-induced lesions as well as their disappearance or persistence after ganciclovir (GCV)-induced suicide. We monitored the alloreactivity of HSV-Tk-transduced T cells after BMT by studying their in vivo distribution and quantity in peripheral blood and in tissues in a WAG/Rij into Brown Norway fully mismatched rat allogeneic BMT model. Genetically modified T cells were quantified in blood and tissues by fluorescence-activated cell sorting, immunohistochemical analysis, and real-time quantitative polymerase chain reaction (PCR) analysis. A significant increase in the number of allogeneic HSV-Tk(+) T cells was found in particular in spleen and lymph nodes and large numbers were found in tongue, skin, and intestines. In blood, an increase in HSV-Tk(+) T cells closely preceded clinical symptoms of GVHD. Real-time quantitative PCR proved to be a fast and accurate tool by which to quantify transduced T cells both in blood and tissues. This enables the study of the in vivo alloreactivity of retrovirus-transduced cells and the response of HSV-Tk-expressing T cells to GCV-induced suicide therapy. Furthermore, we showed the potential use to study specific cause-effect relationships in a broad range of animal and clinical studies involving genetically engineered cells. |
| Databáze: | OpenAIRE |
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