Efficient Introduction of Macromolecules and Oligonucleotides into Brain Capillary Endothelial Cells Using HVJ-Liposomes

Abstract

In this study, we examined the feasibility of introducing macromolecules into cultured mouse brain capillary endothelial cells (MBEC4 cells) by utilizing the hemagglutating virus of Japan (HVJ)-liposomes with fusogenic activity. We used fluorescein isothiocyanate dextran (FITC-Dextran) and FITC-labeled oligodeoxynucleotide (FITC-ODN) as models of a macromolecule and an ODN, respectively. Intracellular fluorescence appeared rapidly after the exposure of MBEC4 cells to FITC-Dextran-containing HVJ-liposomes, and remained detectable for at least 3 days. Only a control level of intracellular fluorescence was seen after treatment with FITC-Dextran alone, FITC-Dextran with empty HVJ-liposomes or FITC-Dextran-containing liposomes without fusogenic activity. In the early phase after administration (0-30 min), the introduction of FITC-Dextran into MBEC4 cells by the HVJ-liposome method resulted in a rapid and time-dependent increase of intracellular fluorescence intensity. Moreover, FITC-ODN was also introduced into MBEC4 cells by the HVJ-liposome method, although FITC-ODN alone was not introduced. These results indicate that the HVJ-liposome method is useful for the efficient introduction of macromolecules, including ODN, into brain capillary endothelial cells.

Transluminal Gene Transfer into Brain Capillary Endothelial Cells In Vivo with HVJ-liposomes

“…6) in comparison with uncoated liposomes or plasmid alone. These results are consistent with our previous findings that HVJ-liposomes could introduce macromolecules into BCECs in vitro (Matsuo et al, 2000) and in vivo (unpublished data) more efficiently than uncoated liposomes. Therefore, the efficient expression of b-Gal gene is at least partly attributable to the effective introduction of the gene by HVJ-liposomes.…”

“…Using the intra-arterial delivery of the HVJliposomes, we previously demonstrated efficient gene transfer into the carotid arterial wall in vivo and arterial endothelial cells in vitro (Yonemitsu et al, 1996). Moreover, we demonstrated that HVJ-liposomes can efficiently transfer macromolecules, such as fluorescein isothiocyanate dextran (FITC-dextran) and FITC-labeled oligodeoxynucleotide (FITC-ODN), into cultured brain capillary endo thelial cells (MBEC4 cells) in vitro (Matsuo et al, 2000).…”

Enhanced gene transfer into brain capillary endothelial cells using Antp-modified DNA-loaded nanoparticles

“…It has been reported that effective delivery of DNA mediated by the unmodified cationic polymers requires 3-6 h exposure [19]. However, our results showed an effective transfection in BCECs exposed to the Antp-modified nanoparticles occurred at 20 min, which was similar to the time-dependent manner of Tat-modified liposomes and HVJ-liposomes [4,20]. Such a rapid increase of intracellular DNA was not achieved by using simplex cationic DNA-polymer complexes, which were taken up via an endocytic mechanism [21].…”

A Review of Nanocarrier-Based CNS Delivery Systems

“…These results indicate that the gene was successfully transferred in vivo from HVJ-liposomes into BCECs, where it was expressed, and the gene product was secreted into the brain. Another study reported the in vitro uptake of HVJ-liposomes encapsulated fluorescein isothiocyanate-labeled dextran (FITC-dextran) and FITC-labeled oligodeoxynucleotide (FITC-ODN) by BCEC's (MBEC4 cells), indicating the utility of these fusogenic vesicles to transport macromolecules and oligonucleotides to BCEC's [60].…”