Source:http://linkedlifedata.com/resource/pubmed/id/10680843
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
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| pubmed:dateCreated |
2000-3-16
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| pubmed:abstractText |
To generate T cell-specific retroviral vectors an scFv phage display library derived from immunized mice was selected for binding to the human T cell line Molt-4/8. The scFv cDNAs recovered from the selected phages were transiently expressed as an N-terminal fusion of the spleen necrosis virus (SNV) transmembrane protein (TM) subunit of the viral envelope protein (Env) in the cell line DSH-cxl, which packages the beta-galactosidase gene into SNV particles. Screening of supernatants from about 150 transfections resulted in the identification of 5 scFvs that mediated efficient transduction of Molt-4/8 cells. Using stable packaging cell lines vector preparations with titers greater than 10(4) EFU/ml on human T cells were obtained. The scFv 7A5 in particular was able to mediate selective transduction of human T cells with high efficiency. Titers of up to 106 EFU/ml were reached on Molt-4/8, Jurkat, and A301 cells, while titers on HeLa cells, TE671 cells, 293T cells, and HT1080 cells were below 102 EFU/ml. Transduction of stimulated primary human peripheral blood cells, which consisted mainly of T cells, was about fivefold more efficient than transduction of B cells. Western blot analysis of supernatant from the 7A5 packaging cells demonstrated incorporation of 7A5-TM into vector particles and indicated proteolytic processing of the coexpressed unmodified TM during particle formation. Binding of bacterially expressed 7A5-scFv to a panel of cell lines correlated well with the transduction results. These data provide the first proof of concept that a general approach can be taken to obtain scFvs able to mediate selective gene transfer into target cells.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical | |
| pubmed:status |
MEDLINE
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| pubmed:month |
Jan
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| pubmed:issn |
1043-0342
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
20
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| pubmed:volume |
11
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
293-303
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| pubmed:dateRevised |
2006-11-15
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| pubmed:meshHeading |
pubmed-meshheading:10680843-Amino Acid Sequence,
pubmed-meshheading:10680843-Animals,
pubmed-meshheading:10680843-Antibodies, Monoclonal,
pubmed-meshheading:10680843-B-Lymphocytes,
pubmed-meshheading:10680843-Blotting, Western,
pubmed-meshheading:10680843-Cell Line,
pubmed-meshheading:10680843-Flow Cytometry,
pubmed-meshheading:10680843-Gene Transfer Techniques,
pubmed-meshheading:10680843-Genetic Vectors,
pubmed-meshheading:10680843-HeLa Cells,
pubmed-meshheading:10680843-Humans,
pubmed-meshheading:10680843-Immunoglobulin Fragments,
pubmed-meshheading:10680843-Leukocytes, Mononuclear,
pubmed-meshheading:10680843-Mice,
pubmed-meshheading:10680843-Mice, Inbred BALB C,
pubmed-meshheading:10680843-Molecular Sequence Data,
pubmed-meshheading:10680843-Peptide Library,
pubmed-meshheading:10680843-Radioimmunoprecipitation Assay,
pubmed-meshheading:10680843-Retroviridae,
pubmed-meshheading:10680843-T-Lymphocytes
|
| pubmed:year |
2000
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| pubmed:articleTitle |
Targeting human T cells by retroviral vectors displaying antibody domains selected from a phage display library.
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| pubmed:affiliation |
Department of Medical Biotechnology, Paul-Ehrlich-Institut, Langen, Germany.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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