Green fluorescent protein-adenoviral construct as a model for transient gene therapy for human cultured keratinocytes in an athymic mouse model.
BACKGROUND
The goal of gene therapy for cultured keratinocyte grafts is to accelerate growth and wound healing following engraftment without producing long-term complications from the delivered gene. We studied a Green Fluorescent Protein-Adenoviral construct (GFP-ADV) to determine the characteristics of gene expression in human cultured keratinocyte grafts.


METHODS
Twelve GFP-ADV grafts and twelve control grafts were transplanted to the flanks of 24 athymic mice. Mouse flanks were monitored with fluorescence-filtered microscopy and, on Day 21, were sectioned and stained with anti-human MHC Class I with H&E counterstaining. Real-time PCR was performed on graft biopsies for adenoviral DNA.


RESULTS
Fluorescence decreased from Days 3 to 5 resulting in no difference between GFP-ADV and control grafts from days 5 to 10. All grafts were positive for human MHC Class I with an epithelial architecture by H&E. Day 21 GFP-ADV grafts were negative for adenoviral DNA.


CONCLUSIONS
The delivered gene was transiently expressed without the persistence of viral DNA, demonstrating the potential of adenoviral gene delivery for the improvement of wound healing without long-term adverse effects to the graft.
Campbell
Chris
C
Department of Surgery, University of North Carolina School of Medicine, 136 Burnett-Womack Building, Chapel Hill, NC 27599-7050, USA.
Hultman
Scott
S
Cairns
Bruce
B
DeSerres
Suzan
S
Meyer
Anthony
A
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001706:Biopsy; D002056:Burns / Q000628:therapy; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured / Q000502:physiology*; D004279:DNA, Viral / Q000032:analysis / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006801:Humans; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D015603:Keratinocytes / Q000166:cytology* / Q000637:transplantation*; D008164:Luminescent Proteins; D051379:Mice; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D016133:Polymerase Chain Reaction; D017768:Skin, Artificial; D014162:Transfection / Q000379:methods*; D014945:Wound Healing
D004279:DNA, Viral; D007202:Indicators and Reagents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins