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    Antigen gene transfer to cultured human dendritic cells using recombinant avipoxvirus vectors.

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    Authors
    Brown, Michael D
    Davies, D H
    Skinner, M A
    Bowen, G
    Hollingsworth, S J
    Mufti, G J
    Arrand, John R
    Stacey, Simon N
    Affiliation
    Cancer Research Campaign, Section of Molecular Genetics, Paterson Institute for Cancer Research, Christie Hospital National Health Service Trust, Manchester, United Kingdom.
    Issue Date
    1999
    
    Metadata
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    Abstract
    Advances in understanding the role of dendritic cells (DCs) as the major antigen (Ag)-presenting cell type of the immune system combined with the recent development of methods for the ex vivo expansion of human DCs have opened the possibility for the transfer of tumor Ags to DCs with a view toward tumor immunotherapy. In this study, we examined the feasibility of Ag transfer to cultured human DCs using the host range-restricted avipoxvirus, fowlpoxvirus (FWPV). FWPV was found to infect and express a lacZ marker gene in a number of mammalian cell lines of fibroblastic, epithelial, and hemopoietic lineage origins. LacZ recombinant FWPV (rFWPV) was found subsequently to infect human DCs that had been cultured ex vivo from peripheral blood monocytes. Using rFWPV containing lacZ under the control of a vaccinia virus (VV) early/late promoter (p7.5K) and a 10 plaque-forming units per cell multiplicity of infection, >80% of cells expressed the lacZ marker gene. Quantitative analysis showed that the level of expression continued to rise for 5 days postinfection, at which point the experiments were terminated. Replication-competent recombinant VV (rVV) was also shown to be capable of transferring the marker gene to primary DC cultures. However, neither rFWPV nor rVV were able to express transgenes under the control of late viral promoters, indicating that both rFWPV and rVV infections are arrested at an early stage in human DCs. Infection of CD83 + DCs by rFWPV was confirmed by double-staining cytochemistry. We conclude that host range-restricted FWPV can be used efficiently to transfer Ag genes to human DCs ex vivo and may have a role in the development of tumor immunotherapy protocols.
    Citation
    Antigen gene transfer to cultured human dendritic cells using recombinant avipoxvirus vectors., 6 (3):238-45 Cancer Gene Ther.
    Journal
    Cancer Gene Therapy
    URI
    http://hdl.handle.net/10541/91418
    DOI
    10.1038/sj.cgt.7700014
    PubMed ID
    10359209
    Type
    Article
    Language
    en
    ISSN
    0929-1903
    ae974a485f413a2113503eed53cd6c53
    10.1038/sj.cgt.7700014
    Scopus Count
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