Long-term expression of human clotting factor IX from retrovirally transduced primary human keratinocytes in vivo

A persistent obstacle that has hampered gene transfer experiments is the short-term nature of transgene expression in vivo. In this article we present evidence for sustained expression from primary human keratinocytes, using the retroviral vector MFG. Primary keratinocytes were transduced in culture...

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Bibliographic Details
Published inHuman gene therapy Vol. 9; no. 8; p. 1187
Main Authors White, S J, Page, S M, Margaritis, P, Brownlee, G G
Format Journal Article
LanguageEnglish
Published United States 20.05.1998
Subjects
Animals
Cell Line
Factor IX - biosynthesis
Gene Transfer Techniques
Genetic Vectors
Humans
Keratinocytes - metabolism
Keratinocytes - transplantation
Mice
Mice, Nude
Moloney murine leukemia virus - genetics
Time Factors
Transduction, Genetic
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Summary:A persistent obstacle that has hampered gene transfer experiments is the short-term nature of transgene expression in vivo. In this article we present evidence for sustained expression from primary human keratinocytes, using the retroviral vector MFG. Primary keratinocytes were transduced in culture with the MFG retroviral vector containing the coding region from factor IX cDNA. Transduced keratinocytes, which secreted on average 830 ng of factor IX/10(6) cells/24 hr in tissue culture, were used to form a bilayered skin equivalent and grafted onto nude mice under a silicone transplantation chamber. Between 0.1 and 2.75 ng of human factor IX per milliliter was found in mouse plasma for more than 1 year, suggesting that keratinocyte stem cells were both transduced and grafted. The results show, for the first time, that long-term expression is obtainable in retrovirally transduced keratinocytes after transplantation.
ISSN:1043-0342
DOI:10.1089/hum.1998.9.8-1187