Production and neurotropism of lentivirus vectors pseudotyped with lyssavirus envelope glycoproteins.

We investigated the production efficiency and the gene transfer capacity in the central nervous system of HIV-1-based vectors pseudotyped with either the G protein of the Mokola lyssaviruses (MK-G), a neurotropic virus causing rabies disease, or the vesiculo-stomatitis G protein (VSV-G). Both envelopes induced syncitia in cell cultures. They were incorporated into vector particles and mature virions were observed by electron microscopy. Vector production was two- to sixfold more efficient with VSV-G than with MK-G. For equivalent amounts of physical particles, vector titration was 5- to 25-fold higher with VSV-G than with MK-G pseudotypes on cultured cells, and in vivo gene expression in mouse brain was more intense. Thus, VSV-G pseudotypes were produced more efficiently and were more infectious than MK-G pseudotypes. Tropism for brain cells was analyzed by intrastriatal injections in rats. Both pseudotypes preferentially transduced neurons (70-90% of transduced cells). Retrograde axonal transport was investigated by instilling vector suspensions in the rat nasal cavity. Both pseudotypes were efficiently transported to olfactive neuron bodies. Thus, although coating HIV-1 particles with rabdhovirus envelope glycoproteins enables them to enter neuronal cells efficiently, pseudotyping is not sufficient to confer the powerful neurotropism of lyssaviruses to lentivirus vectors.

Desmaris N ，Bosch A ，Salaün C ，Petit C ，Prévost MC ，Tordo N ，Perrin P ，Schwartz O ，de Rocquigny H ，Heard JM ... -  《MOLECULAR THERAPY》

Simian immunodeficiency virus vector pseudotypes differ in transduction efficiency and target cell specificity in brain.

Liehl B ，Hlavaty J ，Moldzio R ，Tonar Z ，Unger H ，Salmons B ，Günzburg WH ，Renner M ... -  《GENE THERAPY》

Retinal cell type expression specificity of HIV-1-derived gene transfer vectors upon subretinal injection in the adult rat: influence of pseudotyping and promoter.

Gene therapy, and particularly gene restoration, is currently a great hope for non-curable hereditary retinal degeneration. Clinical applications require a gene transfer vector capable of accurately targeting particular cell types in the retina. To develop such a vector, we compared the expression of a reporter gene after subretinal injections of lentiviral constructs of various pseudotypes and with the transgene expression driven by various promoters. Lentiviral vectors expressing the green fluorescent protein (GFP) under the transcriptional control of cytomegalovirus (CMV), mouse phosphoglycerate kinase (PGK), human elongation factor 1-alpha (EF1alpha), or human rhodopsin (RHO) promoters were pseudotyped by vesicular stomatitis virus (VSV) or Mokola virus envelope proteins. These constructs were injected into the subretinal space of adult rdy rats. GFP expression was analyzed in vivo 1 and 4 weeks after injection by fundus examination. The precise location of transgene expression was then determined by immunohistochemistry and in situ hybridization. Constructs of both vesicular stomatitis virus and Mokola pseudotypes with ubiquitous promoters led to a strong expression of GFP in vivo. Histological studies confirmed the production of GFP in the retinal pigment epithelium (RPE) in most cases. However, only the combination of the VSV pseudotype with the RHO promoter led to GFP production in photoreceptors, and did so in a sporadic manner. Mokola-pseudotyped lentiviral vectors are effective for specific gene transfer to the RPE. Neither VSV- nor Mokola-pseudotyped lentiviral vectors are adequate for efficient gene transfer to photoreceptors of adult rats.

Bemelmans AP ，Bonnel S ，Houhou L ，Dufour N ，Nandrot E ，Helmlinger D ，Sarkis C ，Abitbol M ，Mallet J ... -  《JOURNAL OF GENE MEDICINE》

Production of high-titer human immunodeficiency virus type 1 pseudotyped with vesicular stomatitis virus glycoprotein.

Bartz SR ，Vodicka MA 《METHODS》

Gene transfer in human skin with different pseudotyped HIV-based vectors.

Hachiya A ，Sriwiriyanont P ，Patel A ，Saito N ，Ohuchi A ，Kitahara T ，Takema Y ，Tsuboi R ，Boissy RE ，Visscher MO ，Wilson JM ，Kobinger GP ... -  《GENE THERAPY》