Incorporation of decay-accelerating factor into the baculovirus envelope generates complement-resistant gene transfer vectors.
Baculovirus vectors are an efficient means to deliver genes into hepatocytes in vitro. In experiments that exclude components of the complement system, gene transfer is facilitated. Therefore, the complement system has been defined to represent a potent primary barrier to direct application of baculoviruses in vivo. Here we have genetically manipulated baculoviruses so that the complement-regulatory protein human decay- accelerating factor (DAF) is incorporated into the viral envelope. We found that this modification protected baculovirus vectors against complement-mediated inactivation. Complement-resistant baculovirus vectors were additionally analyzed by immunoblotting and electron microscopy, showing the extent of envelope-incorporated DAF and shape of complement-resistant baculoviruses after exposure to complement. This modified baculovirus vector allowed for an enhanced gene transfer into complement-sufficient neonatal rats in vivo, and thus represents a step in the development of improved alternative viral vectors for gene therapy.
Hüser
A
A
DeveloGen AG Berlin, Robert-Rössle-Str. 10, D-13125 Berlin, Germany.
Rudolph
M
M
Hofmann
C
C
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D018958:CD55 Antigens / Q000235:genetics* / Q000276:immunology; D003167:Complement Activation; D003169:Complement Inactivator Proteins / Q000235:genetics* / Q000276:immunology; D003165:Complement System Proteins / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D015151:Immunoblotting; D008854:Microscopy, Electron; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D014407:Tumor Cells, Cultured; D014760:Viral Fusion Proteins / Q000235:genetics
D018958:CD55 Antigens; D003169:Complement Inactivator Proteins; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins; D003165:Complement System Proteins