In Vivo Gene Transfer Using a Nonprimate Lentiviral Vector Pseudotyped with Ross River Virus Glycoproteins

Abstract

Vectors derived from lentiviruses provide a promising gene delivery system. We examined the in vivo gene transfer efficiency and tissue or cell tropism of a feline immunodeficiency virus (FIV)-based lentiviral vector pseudotyped with the glycoproteins from Ross River Virus (RRV). RRV glycoproteins were efficiently incorporated into FIV virions, generating preparations of FIV vector, which after concentration attain titers up to 1.5 ؋ 10 8 TU/ml. After systemic administration, RRV-pseudotyped FIV vectors (RRV/FIV) predominantly transduced the liver of recipient mice. Transduction efficiency in the liver with the RRV/FIV was ca. 20-fold higher than that achieved with the vesicular stomatitis virus G protein (VSV-G) pseudotype. Moreover, in comparison to VSV-G, the RRV glycoproteins caused less cytotoxicity, as determined from the levels of glutamic pyruvic transaminase and glutamic oxalacetic transaminase in serum. Although hepatocytes were the main liver cell type transduced, nonhepatocytes (mainly Kupffer cells) were also transduced. The percentages of the transduced nonhepatocytes were comparable between RRV and VSV-G pseudotypes and did not correlate with the production of antibody against the transgene product. After injection into brain, RRV/FIV preferentially transduced neuroglial cells (astrocytes and oligodendrocytes). In contrast to the VSV-G protein that targets predominantly neurons, <10% of the brain cells transduced with the RRV pseudotyped vector were neurons. Finally, the gene transfer efficiencies of RRV/FIV after direct application to skeletal muscle or airway were also examined and, although transgene-expressing cells were detected, their proportions were low. Our data support the utility of RRV glycoprotein-pseudotyped FIV lentiviral vectors for hepatocyte-and neurogliarelated disease applications.

Innocuous full-length botulinum neurotoxin targets and promotes the expression of lentiviral vectors in central and autonomic neurons

“…2,3 In the case of enveloped virions, which include lentiviruses, targeted gene delivery can be achieved by incorporating envelope-attachment glycoproteins. 4,5 Importantly, although such interventions enhance the viral selectivity for nerve cells, none render transgene expression neuron specific.…”

A replication competent lentivirus (RCL) assay for equine infectious anaemia virus (EIAV)-based lentiviral vectors

“…First, classical virological approaches, such as focus forming assays, 9 are not suitable due to the use of a variety of heterologous envelope glycoproteins to pseudotype retroviral [9][10][11] and lentiviral [12][13][14][15] vectors. Secondly, the use of EIAV as a positive control virus is inappropriate because the cis-acting sequences and regulatory proteins required for natural replication are missing and because the vectors are so divergent from their parental virus: the only lentiviral gene present in the EIAV packaging system is codon-optimized Gag-Pol, and the components are separated onto different plasmids; together these features are aimed at minimizing the potential for recombination.…”

FIV: from lentivirus to lentivector

“…FIV vectors have been used to target cells in the brain, eye, airway, hematopoietic system, liver, muscle and pancreas [8,15,16,19,[45][46][47][48][49][50][51][52][53][54][55][56][57]. Exceptional results in the central nervous system have been obtained [47].…”