Source:http://linkedlifedata.com/resource/pubmed/id/10022543
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
1999-4-13
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pubmed:abstractText |
Dendritic cells (DCs), the most potent antigen-presenting cells, can be generated from CD34+ hematopoietic stem cells and used for generating therapeutic immune responses. To develop immunotherapy protocols based on genetically modified DCs, we have investigated the conditions for high-level transduction of a large amount of CD34+-derived DCs. Thus, we have used an efficient and clinically applicable protocol for the retroviral transduction of cord blood (CB) or mobilized peripheral blood (MPB) CD34+ cells based on infection with gibbon ape leukemia virus (GALV)-pseudotyped retroviral vectors carrying the nls-LacZ reporter gene. Infected cells have been subsequently cultured under conditions allowing their dendritic differentiation. The results show that using a growth factor combination including granulocyte-macrophage colony-stimulating factor plus tumor necrosis factor alpha plus interleukin 4 plus stem cell factor plus Flt3 ligand, more than 70% of DCs derived from CB or MPB CD34+ cells can be transduced. Semiquantitative PCR indicates that at least two proviral copies per cell were detected. Transduced DCs retain normal immunophenotype and potent T cell stimulatory capacity. Finally, by using a semisolid methylcellulose assay for dendritic progenitors (CFU-DCs), we show that more than 90% of CFU-DCs can be transduced. Such a highly efficient retrovirus-mediated gene transfer into CD34+-derived DCs makes it possible to envision the use of this methodology in clinical trials.
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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
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pubmed:day |
20
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pubmed:volume |
10
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
175-87
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:10022543-Antigens, CD34,
pubmed-meshheading:10022543-Base Sequence,
pubmed-meshheading:10022543-Cell Division,
pubmed-meshheading:10022543-Cell Line,
pubmed-meshheading:10022543-DNA Primers,
pubmed-meshheading:10022543-Dendritic Cells,
pubmed-meshheading:10022543-Fetal Blood,
pubmed-meshheading:10022543-Gene Transfer Techniques,
pubmed-meshheading:10022543-Hematopoietic Stem Cell Mobilization,
pubmed-meshheading:10022543-Humans,
pubmed-meshheading:10022543-Immunophenotyping,
pubmed-meshheading:10022543-Retroviridae,
pubmed-meshheading:10022543-T-Lymphocytes,
pubmed-meshheading:10022543-Transduction, Genetic
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pubmed:year |
1999
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pubmed:articleTitle |
High level of retrovirus-mediated gene transfer into dendritic cells derived from cord blood and mobilized peripheral blood CD34+ cells.
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pubmed:affiliation |
Biologie et Thérapie des Pathologies Immunitaires, ESA CNRS 7087 CERVI, CHU Pitié Salpétrière, Paris, France.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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