Addition of the Human Interferon β Scaffold Attachment Region to Retroviral Vector Backbones Increases the Level ofin VivoTransgene Expression among Progeny of Engrafted Human Hematopoietic Stem Cells
| Autor: | L J Murray, Beth L Hill, Marilyn Travis, Ivan Plavec, E. G. Hanania, Katherine Olsson, Karin Luens-Abitorabi, Sean P. Forestell |
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| Rok vydání: | 2000 |
| Předmět: |
Stromal cell
Transgene Genetic enhancement Genetic Vectors CD34 Antigens CD34 Cell Separation Mice SCID Biology Polymerase Chain Reaction Cell Line Viral vector Mice Transduction Genetic Gene expression Genetics Animals Humans Transgenes Molecular Biology Gene Transfer Techniques Genetic Therapy Interferon-beta Flow Cytometry Hematopoietic Stem Cells Virology Molecular biology Mice Mutant Strains Haematopoiesis Retroviridae Molecular Medicine Moloney murine leukemia virus Stem cell |
| Zdroj: | Human Gene Therapy. 11:2039-2050 |
| ISSN: | 1557-7422 1043-0342 |
| Popis: | Absence of durable high-level expression of transgenes from Moloney murine leukemia (Mo-MuLV) retroviral vectors has been a hurdle in bringing effective gene therapy to the clinic. In this study we have analyzed transgene expression among the progeny of mobilized hematopoietic stem cells (HSCs), comparing Mo-MuLV and mouse stem cell virus (MSCV) vectors, with or without addition of a scaffold attachment region (SAR) from the human interferon beta gene. Retroviral (RV) vector supernatant quality was assessed by comparing NGFR transgene expression by HEL cells, and transgene delivery and expression by CD34(+) cells 72 hr after transduction, using real-time PCR and FACS analysis. This is the first description of the effect of SAR within both Mo-MuLV and MSCV vector backbones on long-term RV transgene expression among in vivo HSC progeny in HSC repopulation assays (SCID-hu bone and NOD/SCID). After transduction of mobilized CD34(+) cells with MSCV-SAR vector, transgene expression was observed among a mean of 10% of donor HSC progeny in the SCID-hu bone (range, 0.6-43%). The predominant effect of SAR was to increase the mean fluorescence intensity (MFI) of transgene expression among HSC progeny in both in vivo bone repopulation models (three- to fourfold), and after long-term stromal cultures (twofold). |
| Databáze: | OpenAIRE |
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