Source:http://linkedlifedata.com/resource/pubmed/id/17235289
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
7
|
| pubmed:dateCreated |
2007-3-20
|
| pubmed:abstractText |
The transfer of T-cell receptor (TCR) genes into primary human T-cells to endow their specificity toward virus-infected and tumor cells is becoming an interesting tool for immunotherapy. TCR-modified T cells are mainly generated by retrovirus-mediated gene transfer. To produce TCR-retrovirus particles, fibroblast packaging cell lines are the most common tool. We constructed two packaging cell lines based on the human suspension T-cell lymphoma line Deltabeta-Jurkat, which lacks endogenous TCRbeta-chains and is therefore unable to express CD3 complexes on the cell surface. After supply of gag-pol (murine leukemia virus (Mo-MLV)) and env (GALV or MLV-10A1) genes, a green fluorescent protein (GFP)-encoding retrovirus vector was transduced into both packaging cell clones, which then stably produced GFP-retroviruses with titers of up to 4 x 10(5) infectious particles (IP)/ml. After transfer of a TCRalpha/beta-encoding retrovirus vector, Deltabeta-Jurkat/GALV and Deltabeta-Jurkat/10A1 cells expressed CD3 molecules on the cell surface. CD3-high expressing packaging cells were enriched by fluorescence-activated cell sorter sorting. In these cells, the CD3 expression level directly correlated with the titer of vector particles. TCR-retroviruses efficiently transduced human T-cell lines and primary T cells. In conclusion, the method allowed the fast and easy generation of high virus titer supernatants for TCR gene transfer.
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| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical | |
| pubmed:status |
MEDLINE
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| pubmed:month |
Apr
|
| pubmed:issn |
0969-7128
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| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
14
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
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| pubmed:pagination |
595-603
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| pubmed:meshHeading |
pubmed-meshheading:17235289-Adoptive Transfer,
pubmed-meshheading:17235289-Antigens, CD3,
pubmed-meshheading:17235289-Blotting, Western,
pubmed-meshheading:17235289-Clone Cells,
pubmed-meshheading:17235289-Flow Cytometry,
pubmed-meshheading:17235289-Gene Expression Regulation, Viral,
pubmed-meshheading:17235289-Gene Therapy,
pubmed-meshheading:17235289-Genetic Engineering,
pubmed-meshheading:17235289-Genetic Vectors,
pubmed-meshheading:17235289-Green Fluorescent Proteins,
pubmed-meshheading:17235289-Humans,
pubmed-meshheading:17235289-Jurkat Cells,
pubmed-meshheading:17235289-Receptors, Antigen, T-Cell,
pubmed-meshheading:17235289-Retroviridae,
pubmed-meshheading:17235289-Reverse Transcriptase Polymerase Chain Reaction,
pubmed-meshheading:17235289-T-Lymphocytes,
pubmed-meshheading:17235289-Transduction, Genetic
|
| pubmed:year |
2007
|
| pubmed:articleTitle |
Suspension packaging cell lines for the simplified generation of T-cell receptor encoding retrovirus vector particles.
|
| pubmed:affiliation |
Max-Delbrück-Center for Molecular Medicine, Berlin, Germany.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|