Abstract:
:Recently, efficient and long-term in vivo gene transfer by recombinant adeno-associated virus type 2 (rAAV) vectors has been demonstrated in a variety of tissues. Further improvement in vector titer and purity will expedite this in vivo exploration and provide preclinical information required for use in human gene therapy. In an effort to obtain higher titers, we constructed a novel AAV helper plasmid which utilizes translational control of AAV Rep genes (J. Li et al., J. Virol. 71:5236-5243, 1997). To address the issue of purity, in this study we report the first rAAV production method which is completely free of adenovirus (Ad) helper virus. The new production system uses a plasmid construct which contains a mini-Ad genome capable of propagating rAAV in the presence of AAV Rep and Cap genes. This construct is missing some of the early and most of the late Ad genes and is incapable of producing infectious Ad. Transfection of 293 cells with the new mini-Ad helper and AAV packaging plasmids results in high-titer rAAV vectors with yields greater than 1,000 transducing units, or 10(5) viral particles per cell. When rAAV vectors were produced by using this production scheme and compared to traditional heat-inactivated rAAV preparations in vitro and in vivo, we observed transduction equivalent to or better than normal levels. The complete removal of infectious Ad from AAV production should facilitate a better understanding of immune response to AAV vectors in vivo, eliminate the need for developing replication-competent Ad assays, and provide a more defined reagent for clinical use.
journal_name
J Viroljournal_title
Journal of virologyauthors
Xiao X,Li J,Samulski RJdoi
10.1128/JVI.72.3.2224-2232.1998subject
Has Abstractpub_date
1998-03-01 00:00:00pages
2224-32issue
3eissn
0022-538Xissn
1098-5514journal_volume
72pub_type
杂志文章abstract::Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), in combination with proteolytic protection assays, has been used to identify the functional epitope on human immunodeficiency virus envelope glycoprotein gp41 for the broadly neutralizing anti-gp41 human monoclonal antibody 2F5. In this protecti...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.75.22.10906-10911.2001
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abstract::Rabies virus (RV) vaccine strain-based vectors show significant promise as potential live-attenuated vaccines against human immunodeficiency virus type 1 (HIV-1). Here we describe a new RV construct that will also likely have applications as a live-attenuated or killed-particle immunogen. We have created a RV containi...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.77.23.12782-12794.2003
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.1.357-364.1995
更新日期:1995-01-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.15.10003-10012.2005
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abstract::Langat virus (LGT), the naturally attenuated member of the tick-borne encephalitis virus (TBEV) complex, was tested extensively in clinical trials as a live TBEV vaccine and was found to induce a protective, durable immune response; however, it retained a low residual neuroinvasiveness in mice and humans. In order to ...
journal_title:Journal of virology
pub_type: 杂志文章
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更新日期:2006-02-01 00:00:00
abstract::Six nonoverlapping peptides of the neuraminidase (NA) glycoprotein of influenza virus A/Puerto Rico/8/34 (H1N1) (PR8 virus) were found to be immunogenic for proliferating T cells when injected into BALB/c mice in Freund adjuvant. T cells elicited by peptide immunization could recognize PR8 virus in vitro. However, onl...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.65.2.672-676.1991
更新日期:1991-02-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.68.11.7083-7091.1994
更新日期:1994-11-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.17.11392-11402.2005
更新日期:2005-09-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.57.3.952-959.1986
更新日期:1986-03-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.10.4645-4651.2000
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.18.8234-8242.2000
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abstract::Herpes simplex virus 1 genome consists of two covalently linked components, L and S, that invert relative to each other to yield four equimolar isomeric populations designated P (prototype), Is (inversion of S component), Il (inversion of L component), and Ils (inversion of L and S components) differing in the orienta...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.59.2.494-499.1986
更新日期:1986-08-01 00:00:00
abstract::An immediate-early protein of murine cytomegalovirus (MCMV), pp89, elicits an immunodominant and protective major histocompatibility complex (MHC) class I Ld-restricted CD8+ T-lymphocyte response. Remarkably, presentation of the naturally processed peptide of pp89, the nonapeptide YPHFMPTNL, is abolished during permis...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.68.1.289-297.1994
更新日期:1994-01-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02152-08
更新日期:2009-03-01 00:00:00
abstract::High-risk human papillomavirus type 16 (HPV-16) and HPV-18 are associated with the majority of human cervical carcinomas, and two viral genes, HPV E6 and E7, are commonly found to be expressed in these cancers. The presence of HPV-16 E7 is sufficient to induce epidermal hyperplasia and epithelial tumors in transgenic ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.8.5905-5914.1997
更新日期:1997-08-01 00:00:00
abstract::Wild-type bacteriophage phie and IS (interference-sensitive) mutants of the related phage SP82G did not productively infect strains of Bacillus subtilis that were lysogenic for temperate phage SPO2. In these abortive infections, the sensitive phages adsorbed to and penetrated the nonpermissive host, phage-directed mac...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.18.3.839-847.1976
更新日期:1976-06-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02323-20
更新日期:2021-01-06 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.67.6.3304-3311.1993
更新日期:1993-06-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.76.11.5315-5325.2002
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.1.3.591-598.1967
更新日期:1967-06-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00722-20
更新日期:2020-08-31 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.3.1984-1991.1997
更新日期:1997-03-01 00:00:00
abstract::Broadly neutralizing antibodies have been isolated that bind the glycan shield of the HIV-1 envelope spike. One such antibody, PGT135, contacts the intrinsic mannose patch of gp120 at the Asn332, Asn392, and Asn386 glycosylation sites. Here, site-specific glycosylation analysis of recombinant gp120 revealed glycan mic...
journal_title:Journal of virology
pub_type: 杂志文章
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更新日期:2015-07-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.61.4.1248-1252.1987
更新日期:1987-04-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
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更新日期:2018-04-27 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.76.24.12845-12854.2002
更新日期:2002-12-01 00:00:00
abstract::Human cytomegalovirus (HCMV) infection has been shown to activate the mTORC1 signaling pathway. However, the phosphorylation of mTORC1 targets is differentially sensitive to the mTORC1 inhibitor rapamycin, and the drug inhibits HCMV replication to a modest extent. Using Torin1, a newly developed inhibitor that targets...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02733-09
更新日期:2010-05-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.75.22.10787-10799.2001
更新日期:2001-11-01 00:00:00
abstract::Members of the Flaviviridae encode a serine proteinase termed NS3 that is responsible for processing at several sites in the viral polyproteins. In this report, we show that the NS3 proteinase of the pestivirus bovine viral diarrhea virus (BVDV) (NADL strain) is required for processing at nonstructural (NS) protein si...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.7.5312-5322.1997
更新日期:1997-07-01 00:00:00