Development of inducible EIAV-based lentiviral vector packaging and producer cell lines

Stewart, Harry E.; Leroux-Carlucci, Marie A; Sion, Christelle J. M; Mitrophanous, Kyriacos; Radcliffe, Pippa A

doi:10.1038/gt.2009.20

Cited by 47 publications

(27 citation statements)

References 40 publications

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“…Only one of the reported inducible HIV-based PCLs, named GPRG, has been proposed for the production of therapeutic vectors for use in clinical trials targeting SCID-Xl1617. Another inducible EIAV-based LV producer cell line has been developed to make therapeutic vectors for use in clinical trials targeting Parkinson's disease1819. However, the scaling-up of inducible systems necessary for clinical-grade LV production is problematic, and additional purification steps of the vector preps to eliminate inducing agents are required.…”

mentioning

confidence: 99%

Construction of stable packaging cell lines for clinical lentiviral vector production

Sanber

Knight

Stephen

et al. 2015

Sci Rep

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Lentiviral vectors are useful experimental tools for stable gene delivery and have been used to treat human inherited genetic disorders and hematologic malignancies with promising results. Because some of the lentiviral vector components are cytotoxic, transient plasmid transfection has been used to produce the large batches needed for clinical trials. However, this method is costly, poorly reproducible and hard to scale up. Here we describe a general method for construction of stable packaging cell lines that continuously produce lentiviral vectors. This uses Cre recombinase-mediated cassette exchange to insert a codon-optimised HIV-1 Gag-Pol expression construct in a continuously expressed locus in 293FT cells. Subsequently Rev, envelope and vector genome expression cassettes are serially transfected. Vector titers in excess of 106 transducing units/ml can be harvested from the final producer clones, which can be increased to 108 TU/ml by concentration. This method will be of use to all basic and clinical investigators who wish to produce large batches of lentiviral vectors.

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mentioning

confidence: 99%

Construction of stable packaging cell lines for clinical lentiviral vector production

Sanber

Knight

Stephen

et al. 2015

Sci Rep

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“…The consequences of human infection with LV derived from non-human primate-and non-primate-LV are unknown and thus safety concerns remain, particularly in association with risks from horizontal and cross-species transmission of any mobilized, recombined chimeric lentivirus. Noteworthy, a gene-based therapy for Parkinson's using EIAV is currently being evaluated in a Phase I/II trial [111,132].…”

Section: Non-human Lentivirusesmentioning

confidence: 99%

State-of-the-Art Lentiviral Vectors for Research Use: Risk Assessment and Biosafety Recommendations

Pauwels¹,

Gijsbers²,

Toelen³

et al. 2009

CGT

108

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Lentiviral vectors (LV) are competent gene transfer vehicles, as used for both research and gene therapy applications, because of their stable integration in non-dividing and dividing cells and long-term transgene expression. Along with our understanding that LV offer solutions for gene therapy, biosafety concerns have uncovered risks due to insertional mutagenesis, the generation of replication competent lentiviruses (RCL) and vector mobilization. Researchers therefore continue to devote significant efforts in designing LV with improved efficacy and biosafety features.The choice of a particular LV system for experimental studies is often driven by functional considerations, including increased productivity and/or transduction efficiency. The design of safer vectors has also directly benefited researchers allowing them to conduct experimental studies with lower risk. Currently, vectors combine improved safety features (that decrease the risk of recombination and vector mobilization) with increased transduction efficiency. Hence, risks associated with the inadvertent transduction of cells of the investigator gain greater importance in assessing the overall risk of these vectors and become an important biosafety concern.This review outlines the different strategies used to improve LV biosafety by comparing state-of-the-art and emerging LV production systems and highlighting biosafety issues that can arise during their contained use. The few existing national and international biosafety recommendations that specifically address the use of LV in research are discussed and recommendations for most common research activities using LV are proposed.

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“…In addition to HIV-derived, other lentiviral vectors have been developed and reported to retain identical features to those of HIV's based, including the ability to transduce nondividing cells, high titers production, and the possibility to be pseudotyped with different envelope glycoproteins. These include lentiviral vectors based on SIV (simian immunodeficiency virus) (Pandya et al 2001;Schnell et al 2000), BIV (bovine immunodeficiency virus) (Matukonis et al 2002;Molina et al 2004), FIV (feline immunodeficiency virus) (Poeschla et al 1998;Saenz and Poeschla 2004) and EAIV (equine infectious anaemia virus) (Balaggan et al 2006;Mitrophanous et al 1999;Stewart et al 2009). Most of non-HIV derived lentiviral vectors have been reported to be tat and sometimes rev independent, thus falling in the 3 rd or 4 th generation of packaging systems.…”

Section: Lentiviral Vectorsmentioning

confidence: 99%

“…At a laboratory scale, transient production by plasmid transfection has been the first choice to cope with the cytotoxic proteins. For larger-scale production purposes, conditional packaging systems have been developed in which the expression of those is under the control of inducible promoters (Broussau et al 2008;Farson et al 2001;Kuate et al 2002;Pacchia et al 2001;Stewart et al 2009). However transient transfection systems are, as discussed above, difficult to scale-up and do not fulfill adequate batch-to-batch variability standards; and, although the clinical trials currently using lentiviral vectors have been provided exclusively with transiently produced batches (Schweizer and Merten 2010), it is unlikely that a transient based systems will be approved when going from clinical to market.…”

Section: Lentiviral Vectorsmentioning

confidence: 99%

Production of Retroviral and Lentiviral Gene Therapy Vectors: Challenges in the Manufacturing of Lipid Enveloped Virus

Rodrigues¹,

Alves²,

Coroadinha³

2011

Viral Gene Therapy

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Development of inducible EIAV-based lentiviral vector packaging and producer cell lines

Cited by 47 publications

References 40 publications

Construction of stable packaging cell lines for clinical lentiviral vector production

Construction of stable packaging cell lines for clinical lentiviral vector production

State-of-the-Art Lentiviral Vectors for Research Use: Risk Assessment and Biosafety Recommendations

Production of Retroviral and Lentiviral Gene Therapy Vectors: Challenges in the Manufacturing of Lipid Enveloped Virus

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