Transgenic expression of the coxsackie/adenovirus receptor enables adenoviral-mediated gene delivery in naive T cells.
The inability to easily and efficiently introduce genes into primary T cells has hampered the investigation of the pathways controlling T cell fate. To enable adenoviral-mediated gene transfer into normal naive T cells, transgenic (Tg) mice expressing the coxsackie/adenovirus receptor (CAR) in their T cell compartment were constructed. Whereas naive T cells are resistant to adenoviral infection, Tg expression of CAR on T cells greatly facilitates adenoviral-mediated gene expression ex vivo, in vivo, and in differentiated T helper cells. Thus we have developed a technology for efficient gene delivery to naive T cells. By using adenoviral vectors encoding specific inhibitors, we show that G1 cyclin-dependent kinase, NF-kappaB, and caspase activities are required for the proliferation of primary T cells. In addition, by expressing Bcl-x(L) protein at a level that closely approximates mitogen-induced levels, we demonstrate that Bcl-x(L) expression is sufficient to account for mitogen-mediated survival of primary T cells. Thus, adenoviral-mediated gene delivery to CAR Tg T cells should be useful for the analysis of many genes controlling T cell fate.
Wan
Y Y
YY
Departments of Biochemistry and Molecular Genetics and Pediatrics, and Microbiology, Program in Molecular Biology, University of Colorado Health Sciences Center, BRB802, 4200 East Ninth Avenue, Denver, CO 80262, USA.
Leon
R P
RP
Marks
R
R
Cham
C M
CM
Schaack
J
J
Gajewski
T F
TF
DeGregori
J
J
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002455:Cell Division; D004770:Enterovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D011991:Receptors, Virus / Q000235:genetics*; D011995:Recombination, Genetic; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D014161:Transduction, Genetic
D011991:Receptors, Virus