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 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.
White
S J
SJ
Chemical Pathology Unit, Sir William Dunn School of Pathology, University of Oxford, UK.
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S M
SM
Margaritis
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P
Brownlee
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GG
D000818:Animals; D002460:Cell Line; D005164:Factor IX / Q000096:biosynthesis*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism* / Q000637:transplantation; D051379:Mice; D008819:Mice, Nude; D008979:Moloney murine leukemia virus / Q000235:genetics*; D013997:Time Factors; D014161:Transduction, Genetic
D005164:Factor IX