Baculovirus-mediated periadventitial gene transfer to rabbit carotid artery

Recombinant Autographa californica multiple nuclear polyhedrosis viruses (A cMNPV) have recently been shown to transduce mammalian cells in vitro. Sinc e baculoviruses offer many advantages over viruses currently used in gene t herapy we have tested them for in vivo gene transfer by constructing a bacu lovirus bearing a nuclear targeted beta-galactosidase marker gene (LacZ) un der a CMV promoter. Both rabbit aortic smooth muscle cells (RAASMC) and hum an EGV-304 cells were susceptible to LacZ-baculovirus transduction. Transge ne expression was evaluated in vivo by applying 1 x 10(9) p.f.u. of LacZ-ba culoviruses or LacZ-adenoviruses in a silastic collar placed around rabbit carotid arteries in the absence of contact with blood components. As a resu lt, baculoviruses led to transgene expression in adventitial cells in rabbi t carotid arteries with efficiency comparable to adenoviruses. The beta-gal actosidase gene expression was transient staying at a high level for 1 week but disappearing at the 14 day time-point. The arterial structure and endo thelium remained intact in the baculovirus-transduced arteries, but macroph age-specific immunostaining defected signs of inflammation comparable to ad enoviruses. Baculoviruses are thus able to mediate transient gene transfer in vive and may become useful tools for gene therapy.