Expression of vhs and VP16 during HSV-1 helper virus-free amplicon packaging enhances titers

Bowers, WJ; Howard, D. H.; Brooks, AI; Halterman, MW; Federoff, H J

doi:10.1038/sj.gt.3301340

Cited by 54 publications

(41 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…[47][48][49][50] This packaging strategy requires the cotransfection of eucaryotic cells that are receptive to HSV propagation (ie, BHK, Vero) with packaging-incompetent HSV genomic DNA, amplicon DNA, and any accessory HSV genes shown to enhance amplicon titers. 51 Crude vector lysates are then purified by a series of ultracentrifugation steps and titered by expression or transduction-based methodologies. 52 The titers obtained from helper virus-free amplicon packaging typically range from 10 7 to 10 8 expressing virus particles/ml.…”

Section: Hsv Amplicon Vectorsmentioning

confidence: 99%

Immune responses to replication-defective HSV-1 type vectors within the CNS: implications for gene therapy

2003

View full text Add to dashboard Cite

Section: Hsv Amplicon Vectorsmentioning

confidence: 99%

Immune responses to replication-defective HSV-1 type vectors within the CNS: implications for gene therapy

2003

View full text Add to dashboard Cite

“…Recombinant adenoviruses containing HA-Pincher and HAPincherG68E (7), T7-RacN17, EGFR, ETrkB, EGFR-GFP, Rab5-GFP, Rab7-GFP, or RacV12-GFP were made using the Ad-Easy system (Stratagene). Recombinant adenoviruses containing HSVs containing PincherG68E or GFP were packaged into helper virus-free amplicon particles (29).…”

Section: Methodsmentioning

confidence: 99%

Trk retrograde signaling requires persistent, Pincher-directed endosomes

Philippidou

Valdez

Akmentin

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Target-derived neurotrophins use retrogradely transported Trk-signaling endosomes to promote survival and neuronal phenotype at the soma. Despite their critical role in neurotrophin signaling, the nature and molecular composition of these endosomes remain largely unknown, the result of an inability to specifically identify the retrograde signaling entity. Using EGF-bound nanoparticles and chimeric, EGF-binding TrkB receptors, we elucidate Trk-endosomal events involving their formation, processing, retrograde transport, and somal signaling in sympathetic neurons. By comparing retrograde endosomal signaling by Trk to the related but poorly neuromodulatory EGF-receptor, we find that Trk and EGF-receptor endosomes are formed and processed by distinct mechanisms. Surprisingly, Trk and EGF-receptors are both retrogradely transported to the soma in multivesicular bodies. However, only the Trk-multivesicular bodies rely on Pincher-dependent macroendocytosis and processing. Retrograde signaling through Pincher-generated Trk-multivesicular bodies is distinctively refractory to signal termination by lysosomal processing, resulting in sustained somal signaling and neuronal gene expression.

show abstract

“…HSV-1 amplicons have a transgene capacity sufficient to carry the NR2D cDNA and the coexpressed GFP reporter gene (Arvanian et al, 2004). HSV-1-gfpmediated gene expression peaks during the first 24 -48 h (Bowers et al, 2000(Bowers et al, , 2001. The motoneuron infection was determined by using double immunolabeling with peripherin (motoneurons marker, green) and with GFP (red).…”

Section: Spinal Cord Injury and Treatment Deliverymentioning

confidence: 99%

“…Packaging of helper virus-free amplicon vector stocks and subsequent virus purification and determination of amplicon virus titers via both expression-and transduction-based methodologies were performed as previously described (Bowers et al, 2000(Bowers et al, , 2001. Vectors carried genes for either NR2D (HSV-NR2D) or ␤-galactosidase (HSVlac), as well as the reporter gene green fluorescent protein (GFP).…”

Section: Hsv Amplicon Vector Construction and Packagingmentioning

confidence: 99%

Chondroitinase ABC Combined with Neurotrophin NT-3 Secretion and NR2D Expression Promotes Axonal Plasticity and Functional Recovery in Rats with Lateral Hemisection of the Spinal Cord

García-Alías¹,

Petrosyan²,

Schnell³

et al. 2011

J. Neurosci.

View full text Add to dashboard Cite

Elevating spinal levels of neurotrophin NT-3 (NT3) while increasing expression of the NR2D subunit of the NMDA receptor using a HSV viral construct promotes formation of novel multisynaptic projections from lateral white matter (LWM) axons to motoneurons in neonates. However, this treatment is ineffective after postnatal day 10. Because chondroitinase ABC (ChABC) treatment restores plasticity in the adult CNS, we have added ChABC to this treatment and applied the combination to adult rats receiving a left lateral hemisection (Hx) at T8. All hemisected animals initially dragged the ipsilateral hindpaw and displayed abnormal gait. Rats treated with ChABC or NT3/HSV-NR2D recovered partial hindlimb locomotor function, but animals receiving combined therapy displayed the most improved body stability and interlimb coordination [Basso-Beattie-Bresnahan (BBB) locomotor scale and gait analysis]. Electrical stimulation of the left LWM at T6 did not evoke any synaptic response in ipsilateral L5 motoneurons of control hemisected animals, indicating interruption of the white matter. Only animals with the full combination treatment recovered consistent multisynaptic responses in these motoneurons indicating formation of a detour pathway around the Hx. These physiological findings were supported by the observation of increased branching of both cut and intact LWM axons into the gray matter near the injury. ChABC-treated animals displayed more sprouting than control animals and those receiving NT3/HSV-NR2D; animals receiving the combination of all three treatments showed the most sprouting. Our results indicate that therapies aimed at increasing plasticity, promoting axon growth and modulating synaptic function have synergistic effects and promote better functional recovery than if applied individually.

show abstract

Expression of vhs and VP16 during HSV-1 helper virus-free amplicon packaging enhances titers

Cited by 54 publications

References 32 publications

Immune responses to replication-defective HSV-1 type vectors within the CNS: implications for gene therapy

Immune responses to replication-defective HSV-1 type vectors within the CNS: implications for gene therapy

Trk retrograde signaling requires persistent, Pincher-directed endosomes

Chondroitinase ABC Combined with Neurotrophin NT-3 Secretion and NR2D Expression Promotes Axonal Plasticity and Functional Recovery in Rats with Lateral Hemisection of the Spinal Cord

Contact Info

Product

Resources

About