Regular Article
Uptake and Gene Expression of Naked Plasmid DNA in Cultured Brain Microvessel Endothelial Cells☆

https://doi.org/10.1006/bbrc.1998.8334Get rights and content

Abstract

Cellular uptake and gene expression of plasmid DNA and its cationic liposome complexes were studied using primary cultures of bovine brain microvessel endothelial cells (BMEC) developed as an in vitro model of the blood-brain barrier. An avid association of naked plasmid DNA with the BMEC monolayer was observed at 37°C, which is comparable to that of the DNA/liposome complex.The cellular association significantly decreased at low temperature (4°C). The binding at 4°C was saturable and significantly inhibited by polyanions involving polyinosinic acid and dextran sulfate, typical ligands for the macrophage scavenger receptors, but not by polycytidylic acid or in the presence of EDTA. Unexpectedly, a significant gene expression in the BMEC was obtained by transfection with naked plasmid DNA although the expression level was lower than that obtained by plasmid DNA/cationic liposome complex. Taken together, cultured capillary endothelial cells derived from the brain are able to take up naked plasmid DNA via a scavenger receptor like-mediated mechanism for polyanions and gene expression in the cells takes place.

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    ☆

    Abbreviations: BBB, blood-brain barrier; BMEC, brain microvessel endothelial cells; HUVEC, human umbilical vein endothelial cells; pCMV-CAT, plasmid DNA encoding chloramphenicol acetyltransferase under the control of cytomegalovirus promoter; pCMV-Luc, plasmid DNA encoding firefly luciferase under the control of cytomegalovirus promoter; Poly-I, polyinosinic acid; Poly-C, polycytidylic acid; DS, dextran sulfate

    1

    To whom correspondence should be addressed: Mitsuru Hashida, Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606-01, Japan. Fax: 81-75-753-4575. E-mail:[email protected].

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