Efficient gene transfer into human hepatocytes by baculovirus vectors.

Abstract

Viral vectors are the most efficient tools for gene delivery, and the search for tissue-specific infecting viruses is important for the development of in vivo gene therapy strategies. The baculovirus Autographa californica nuclear polyhedrosis virus is widely used as a vector for expression of foreign genes in insect cells, and its host specificity is supposed to be restricted to arthropods. Here we demonstrate that recombinant A. californica nuclear polyhedrosis virus is efficiently taken up by human hepatocytes via an endosomal pathway. High-level reporter gene expression from heterologous promoters was observed in human and rabbit hepatocytes in vitro. Mouse hepatocytes and some other epithelial cell types are targeted at a considerably lower rate. The efficiency of gene transfer by baculovirus considerably exceeds that obtained by calcium phosphate or lipid transfection. These properties of baculovirus suggest a use for it as a vector for liver-directed gene transfer but highlight a potential risk in handling certain recombinant baculoviruses.

Role of paracellular junction complexes in baculovirus-mediated gene transfer to nondividing rat hepatocytes

“…Many different gene delivery methods are available including cationic lipid-based DNA transfection [44][45][46][47][48][49] and retroviral and adenoviral gene transfer vectors; [50][51][52][53][54][55] however, the use of these methods for hepatocytes has not been effective because hepatocytes do not divide, are highly susceptible to toxicity, and transfection efficiencies in hepatocytes are extremely low (HC Isom, unpublished data). 11 Previous studies have reported baculovirus-mediated gene delivery to primary hepatocytes. 6,11,12 In these studies, baculovirus-mediated gene delivery was highly efficient (470%) and homogeneous.…”

“…11 Previous studies have reported baculovirus-mediated gene delivery to primary hepatocytes. 6,11,12 In these studies, baculovirus-mediated gene delivery was highly efficient (470%) and homogeneous. However, these studies were carried out using cells in short-term culture conditions and as such, the hepatocytes were poorly differentiated and do not have intact paracellular junctions.…”

“…2,5 The baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) can efficiently deliver genes to numerous mammalian cell types including hepatic cells. [6][7][8][9][10][11][12][13][14] Recently, we have characterized baculovirusmediated gene delivery into long-term primary rat hepatocytes maintained in a chemically defined medium supplemented with DMSO. 14 Baculovirus-mediated gene transfer in this culture system was dose-dependent and restricted to peripheral cells of hepatocellular islands when extracellular calcium was present during baculovirus infection.…”

Truncated vesicular stomatitis virus G protein improves baculovirus transduction efficiency in vitro and in vivo

“…To study this hypothesis, the transduction efficiency of HeLa, SKOV-3, BT4C, HepG2, EAHY and 293T cells was determined with the VSV-GED virus and control virus using MOIs ranging from 10 to 1000. 20,21 On the contrary, EAHY cell line has been reported to be poorly susceptible to baculovirus transduction. 22 Transduction efficiency of HeLa, SKOV-3, BT4C and 293T settles between HepG2 and EAHY.…”

Lentiviral vectors pseudotyped with baculovirus gp64 efficiently transduce mouse cells in vivo and show tropism restriction against hematopoietic cell types in vitro

“…16 Early studies showed that baculovirus vectors could efficiently transduce human hepatocyte cell lines and explanted human liver tissue. 17,18 Subsequently, it was demonstrated that baculovirus vectors could enter a broader variety of human cell types. [19][20][21] We set out to determine whether gp64 could be used to pseudotype other viral vectors and confer high infectivity in cells of hepatic origin, which are primary targets for many genetherapy applications.…”