Hemagglutinating Virus of Japan (HVJ) Envelope Vector as a Versatile Gene Delivery System

Abstract

We have developed a simple method for converting the lipid envelope of an inactivated virus to a gene transfer vector. Hemagglutinating virus of Japan (HVJ; Sendai virus) envelope vector was constructed by incorporating plasmid DNA into inactivated HVJ particles. This HVJ envelope vector introduced plasmid DNA efficiently and rapidly into various cell lines, including cancer cells and several types of primary cell culture. Efficiency of gene transfer was greatly enhanced by protamine sulfate and centrifugation. Fluorescein isothiocyanate-labeled oligodeoxynucleotides (FITC-ODN) were also delivered to cells at > 95% efficiency. When HVJ envelope vector was injected into organs directly, reporter gene expression was observed in organs including liver, brain, skin, uterus, tumor masses, lung, and eye. When HVJ envelope vector containing luciferase gene was injected into mouse tail vein, luciferase gene expression was detected primarily in spleen. FITC-ODN were also delivered to spleen cells by intravenous injection of HVJ envelope. These results suggest that HVJ envelope vector will be useful for both ex vivo and in vivo gene therapy experiments.

Ring-Sp1 decoy oligonucleotide effectively suppresses extracellular matrix gene expression and fibrosis of rat kidney induced by unilateral ureteral obstruction

“…It has been reported that a highly effective HVJ-liposome gene transfer technique introduced decoy ODN mainly into renal interstitial fibroblasts. 40 Together with the present results, this suggests that this strategy specifically suppresses Sp1-mediated transcription in the interstitial areas of the targeted kidney, and thus can be used as an effective therapy for tubulointerstitial fibrosis caused in nephritic diseases.…”

“…40 Briefly, egg yolk phosphatidylcholine (Sigma), dioleolyphosphatidylethanolamine (Avanti Polar Lipid, Birmingham, AL, USA), egg yolk sphingomyelin (Sigma), bovine brain phosphatidylserine (Sigma), and cholesterol (Sigma) were mixed in a molar ratio of 16. …”

In vivo transfection of a cis element ‘decoy’ against signal transducers and activators of the transcription 6 (STAT6) binding site ameliorates the response of contact hypersensitivity

“…The subcutaneous injection of STAT6 decoy ODN with HVJ-E after sensitization exhibited a significant inhibitory effect on both STAT6 binding to nucleus and the late-phase response in line with Kaneda's report. 27 A histopathological analysis revealed a tremendous reduction in both the infiltration of eosinophils, mast cells, and especially mast cell degradation in single or repeated challenged skin in STAT6 decoy ODN-transfected mice. STAT6 signaling is essential for the hapten-induced increase in Th2 cytokines production in vivo.…”

“…27 The virus was inactivated by b-propiolactone (0.0075-0.001%) treatment or by UV irradiation. In both cases, the virus preparation lost the ability to replicate.…”

“…The aliquots of the virus (3 Â 10 10 particles/1.5 ml tube) were centrifuged (18 500 g, 15 min) at 41C, and the viral pellet was stored at À201C. 27 The inactivated virus suspension (10 000 haemagglutinating activity units (HAU)) was mixed with STAT6 decoy OND or scrambled decoy ODN (800 mg of DNA), 8 ml of 3% Triton/TE and balanced salt solution (BSS; 10 nM Tris-HCl (pH 7.5), 137 mM NaCl and 5.4 mM KCl)) to yield a final volume of 100 ml. The mixture was centrifuged at 18 500 g for 15 min at 41C.…”