title: Intracellular tracking of protamine/antisense oligonucleotide nanoparticles and their inhibitory effect on HIV-1 transactivation.
abstract: Membrane transport of antisense oligonucleotides (ODN) is an inefficient process which requires special carriers for their intracellular delivery. We have developed a delivery system for AS-ODN and their phosphorothioate analogues (AS-PTO) directed against human immunodeficiency virus type 1 (HIV-1) tat mRNA for efficient transfection of HIV-1 target cells. Protamine was used to complex AS-ODN and AS-PTO to form nanoparticles with diameters of about 180 nm and surface charges in the range of -18 to +30 mV. Cellular uptake of these nanoparticles was significantly enhanced compared to naked oligonucleotides. A double labeling technique with fluorescently tagged protamine and AS-ODN was used to follow the intracellular fate of the nanoparticles. Protamine/AS-ODN nanoparticles showed release of the antisense compound leading to specific inhibition of tat mediated HIV-1 transactivation. In contrast, protamine/AS-PTO complexes were stable over 72 h, and failed to release AS-PTO. These results demonstrate that protamine/AS-ODN nanoparticles are useful for future therapeutical application to inhibit viral gene expression.
authors.affiliation: Department of Virology and Cell Biology, Chemotherapeutisches Forschungsinstitut Georg-Speyer-Haus, Paul-Ehrlich-Strasse 42-44, D-60596 Frankfurt am Main, Germany.
authors.forename: Norbert
authors.initials: N
authors.lastname: Dinauer
authors.forename: Dirk
authors.initials: D
authors.lastname: Lochmann
authors.forename: Ilhan
authors.initials: I
authors.lastname: Demirhan
authors.forename: Abdellatif
authors.initials: A
authors.lastname: Bouazzaoui
authors.forename: Andreas
authors.initials: A
authors.lastname: Zimmer
authors.forename: Angelika
authors.initials: A
authors.lastname: Chandra
authors.forename: Jörg
authors.initials: J
authors.lastname: Kreuter
authors.forename: Hagen
authors.initials: H
authors.lastname: von Briesen
mesh_terms: D002470:Cell Survival / Q000187:drug effects; D015696:Gene Products, tat / Q000737:chemistry / Q000378:metabolism; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D019169:Jurkat Cells; D008027:Light; D008863:Microspheres; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D010316:Particle Size; D011479:Protamines / Q000008:administration & dosage / Q000493:pharmacokinetics / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013499:Surface Properties; D015533:Transcriptional Activation / Q000187:drug effects*; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D015696:Gene Products, tat; D016376:Oligonucleotides, Antisense; D011479:Protamines; D012333:RNA, Messenger; D054322:tat Gene Products, Human Immunodeficiency Virus