Systematic Determination of the Packaging Limit of Lentiviral Vectors

Kumar, Mukesh; Keller, Brian C.; Makalou, Ndeye; Sutton, Richard E.

doi:10.1089/104303401753153947

Cited by 400 publications

(305 citation statements)

References 16 publications

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“…Although considerable progress has been made with regard to spatial control of gene delivery to cells, transient gene expression and poor transfection/transduction efficiencies may not allow sufficient control of cell phenotype and tissue development. LV represents an attractive option for efficient and sustained gene delivery, in addition to other advantageous properties including high packaging capacity (>10 kb), low immunogenicity, low cytotoxicity, and broad tropism (56)(57)(58)(59). The development of self-inactivating vectors has addressed the serious safety concerns of earlier generations of viral systems (60) and made LV an attractive option for clinical use that overcomes the shortcomings of many alternative vector systems.…”

Section: Discussionmentioning

confidence: 99%

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Brunger

Huynh

Guenther

et al. 2014

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

116

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The ability to develop tissue constructs with matrix composition and biomechanical properties that promote rapid tissue repair or regeneration remains an enduring challenge in musculoskeletal engineering. Current approaches require extensive cell manipulation ex vivo, using exogenous growth factors to drive tissue-specific differentiation, matrix accumulation, and mechanical properties, thus limiting their potential clinical utility. The ability to induce and maintain differentiation of stem cells in situ could bypass these steps and enhance the success of engineering approaches for tissue regeneration. The goal of this study was to generate a self-contained bioactive scaffold capable of mediating stem cell differentiation and formation of a cartilaginous extracellular matrix (ECM) using a lentivirus-based method. We first showed that poly-L-lysine could immobilize lentivirus to poly(e-caprolactone) films and facilitate human mesenchymal stem cell (hMSC) transduction. We then demonstrated that scaffoldmediated gene delivery of transforming growth factor β3 (TGF-β3), using a 3D woven poly(e-caprolactone) scaffold, induced robust cartilaginous ECM formation by hMSCs. Chondrogenesis induced by scaffold-mediated gene delivery was as effective as traditional differentiation protocols involving medium supplementation with TGF-β3, as assessed by gene expression, biochemical, and biomechanical analyses. Using lentiviral vectors immobilized on a biomechanically functional scaffold, we have developed a system to achieve sustained transgene expression and ECM formation by hMSCs. This method opens new avenues in the development of bioactive implants that circumvent the need for ex vivo tissue generation by enabling the long-term goal of in situ tissue engineering.biomaterials | regenerative medicine | genetic engineering | gene therapy | chondrocyte

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Section: Discussionmentioning

confidence: 99%

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Brunger

Huynh

Guenther

et al. 2014

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

116

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“…Edited sequences were then aligned to reference sequences representative of all HIV-1 subtypes obtained from the Los Alamos database (http://hivweb.lanl.gov) in ClustalW (Thompson et al 1994). Aligned sequences from env and pol genes were subjected to phylogenetic inference through the neighborjoining method and Kimura 2-parameter model of the MEGA 2.1 package (Kumar et al 2001) for assignment of HIV-1 subtype. The genotypic interpretation of antiretroviral drug resistant mutations in PR and RT was carried out by electronic submission to the Stanford database (http://hivdb.stanford.edu).…”

Section: Seroreactivitymentioning

confidence: 99%

Anti-human immunodeficiency virus type 1 humoral immune response and highly active antiretroviral treatment

Bongertz

Ouverney

Fernandez

et al. 2007

Mem. Inst. Oswaldo Cruz

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Highly active antiretroviral treatment (HAART) of human immunodeficiency type 1 (HIV-1) infection is very effective in controlling infection, but elimination of viral infection has not been achieved as yet, and upon treatment interruption an immediate rebound of viremia is observed. A combination of HAART with an immune stimulation might allow treatment interruption without this rebounding viremia, as the very low viremias observed with successful HAART may be insufficient to permit maintenance of a specific anti-HIV-1 immune response. The objective of this study was to compare the humoral immune response of individuals undergoing successful HAART (NF=no failure) with that of individuals with evidence of failure of therapy (FT) and to verify if the viremia peaks observed in individuals with therapy failure would act as a specific stimulus for the humoral anti-HIV-1 immune response. Antibodies binding to gp120 V3 genotype consensus peptides were more frequently observed for FT, mainly against peptides corresponding to sequences of genotypes prevalent in the Rio de Janeiro city area, B and F. HIV-1 neutralization of HIV-1 IIIB and of four primary isolates from Rio de Janeiro was less frequently observed for plasma from the NF than the FT group, but this difference was more expressive when plasma from individuals with detectable viremia were compared to that of individuals with undetectable viral loads in the year before sample collection. Although statistically significant differences were observed only in some specific comparisons, the study indicates that presence of detectable viremia may contribute to the maintenance of a specific anti-HIV-1 humoral immune response.Key words: antibodies -seroreactivity -neutralization -antiretroviral treatment -treatment failure Anti-HIV highly active antiretroviral treatment (HAART) reduces HIV in peripheral blood and reverts the characteristic immunodeficiency, and has benefited patients increasing individual survival at least 13-14 years (Vermund et al. 2006). However, no elimination of viral infection has been achieved, and the antiretroviral drugs will probably have to be taken throughout the life of the patient. Attempts to stimulate a potent specific anti-HIV-1 immune response to permit control of viral replication. After HAART has reduced viremia to undetectable levels, have so far been less than successful. During successful HAART, a decline in antibody response is generally observed (Fournier et al. 2002, Devito et al. 2006, with cases of seroreversion being described (Amor et al. 2006) and the extent of immune reconstitution in HAART treated individuals is controversial, apparently depending on the immune status of the individuals at the start of therapy (Cheonis et al. 2005). However, the benefits of HAART are evident even when treatment apparently fails, leading neither to a complete control of viremia nor to a complete reconstitution in CD4 T lymphocytes (Brígido et al. 2004, Kovacs et al. 2005).During successful anti-HIV therapy, a reduction in virus specific ce...

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“…LV have a large packaging capacity (410 kb transgene cassette 1,2 ) and envelope glycoprotein pseudotyping achieves broad or specific tropism as necessary. These vectors are also associated with low immunogenicity.…”

Section: Introductionmentioning

confidence: 99%

Integrase-defective lentiviral vectors: progress and applications

2009

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Lentiviral vectors (LVs) offer the advantages of a large packaging capacity, broad cell tropism or specific cell-type targeting through pseudotyping, and long-term expression from integrated gene cassettes. However, transgene integration carries a risk of disrupting gene expression through insertional mutagenesis and may not be required for all applications. A non-integrating LV may be beneficial in cases in which transient gene expression is desired. Several recent publications outline the development and initial biological characterization of such vectors. Here, we discuss the potential applications and new directions for the development of integration-defective LVs.

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Systematic Determination of the Packaging Limit of Lentiviral Vectors

Cited by 400 publications

References 16 publications

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

Anti-human immunodeficiency virus type 1 humoral immune response and highly active antiretroviral treatment

Integrase-defective lentiviral vectors: progress and applications

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