title: Tropism, intracerebral distribution, and transduction efficiency of HIV- and SIV-based lentiviral vectors after injection into the mouse brain: a qualitative and quantitative in vivo study.
abstract: Lentiviruses are suitable to transfer potential therapeutic genes into non-replicating cells such as neurons, but systematic in vivo studies on transduction of neural cells within the complete brain are missing. We analysed the distribution of transduced cells with respect to brain structure, virus tropism, numbers of transduced neurons per brain, and influence of the Vpx or Vpr accessory proteins after injection of vectors based on SIVsmmPBj, HIV-2, and HIV-1 lentiviruses into the right striatum of the mouse brain. Transduced cells were found ipsilaterally around the injection canal, in corpus striatum and along corpus callosum, irrespective of the vector type. All vectors except HIV-2SEW transduced also single cells in the olfactory bulb, hippocampus, and cerebellum. Vector HIV-2SEW was the most neuron specific. However, vectors PBjSEW and HIV-1SEW transduced more neurons per brain (means 41,299 and 32,309) than HIV-2SEW (16,102). In the presence of Vpx/Vpr proteins, HIV-2SEW(Vpx) and HIV-1SEW(Vpr) showed higher overall transduction efficiencies (30,696 and 27,947 neurons per brain) than PBjSEW(Vpx) (6636). The distances of transduced cells from the injection canal did not differ among the viruses but correlated positively with the numbers of transduced neurons. The presence of Vpx/Vpr did not increase the numbers of transduced neurons. Parental virus type and the vector equipment seem to influence cellular tropism and transduction efficiency. Thus, precision of injection and choice of virus pseudotype are not sufficient when targeted lentiviral vector transduction of a defined brain cell population is required.
authors.affiliation: Department of Pathobiology, Institute of Anatomy, Histology and Embryology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210, Vienna, Austria. juraj.hlavaty@vetmeduni.ac.at.
authors.forename: Juraj
authors.initials: J
authors.lastname: Hlavatý
authors.affiliation: Faculty of Applied Sciences, European Centre of Excellence NTIS, University of West Bohemia in Pilsen, Univerzitní 8, 306 14, Pilsen, Czech Republic.
authors.forename: Zbyněk
authors.initials: Z
authors.lastname: Tonar
authors.affiliation: Department of Medicinal Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Renner
authors.affiliation: Department of Medicinal Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Panitz
authors.affiliation: Institute of Virology, University of Veterinary Medicine, Vienna, Austria.
authors.forename: Helga
authors.initials: H
authors.lastname: Petznek
authors.affiliation: Department of Medicinal Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schweizer
authors.affiliation: Department of Medicinal Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Schüle
authors.affiliation: Department of Medicinal Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
authors.forename: Björn-Philipp
authors.initials: BP
authors.lastname: Kloke
authors.affiliation: Institute of Medical Biochemistry, University of Veterinary Medicine, Vienna, Austria.
authors.forename: Rudolf
authors.initials: R
authors.lastname: Moldzio
authors.affiliation: Department of Pathobiology, Institute of Anatomy, Histology and Embryology, University of Veterinary Medicine Vienna, Veterinärplatz 1, 1210, Vienna, Austria.
authors.forename: Kirsti
authors.initials: K
authors.lastname: Witter
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism / Q000821:virology*; D002478:Cells, Cultured; D005260:Female; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics / Q000493:pharmacokinetics*; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D015498:HIV-2 / Q000235:genetics / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D011247:Pregnancy; D036301:Qualitative Research; D015302:Simian Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism*; D014161:Transduction, Genetic / Q000379:methods*; D056189:Viral Tropism
keywords: Double immunofluorescence; EGFP; Gene transfer; Stereology; Vpr; Vpx