Efficient lentiviral transduction of human cord blood CD34(+) cells followed by their expansion and differentiation into dendritic cells

Citation
M. Oki et al., Efficient lentiviral transduction of human cord blood CD34(+) cells followed by their expansion and differentiation into dendritic cells, EXP HEMATOL, 29(10), 2001, pp. 1210-1217
Citations number
37
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Cardiovascular & Hematology Research
Journal title
EXPERIMENTAL HEMATOLOGY
ISSN journal
0301-472X → ACNP
Volume
29
Issue
10
Year of publication
2001
Pages
1210 - 1217
Database
ISI
SICI code
0301-472X(200110)29:10<1210:ELTOHC>2.0.ZU;2-S
Abstract
Objective. To support immune reconstitution after cord blood transplantatio n, immunotherapy using gene-modified dendritic cells (DCs), the most potent antigen-presenting cells, can be a powerful strategy for preventing infect ion and recurrence. To investigate the applicability of lentiviral vector-t ransduced DCs compared to retroviral vectors, we transduced umbilical cord blood (CB) CD34(+) cells, then expanded and differentiated them into DCs. Materials and Methods. We transduced CB CD34(+) cells by vesicular stomatit is virus G-protein pseudotyped self-inactivating lentiviral vector or retro viral vectors carrying the enhanced green fluorescent protein gene. The cel ls were expanded in the stroma-dependent culture system and transferred to the culture condition for developing DCs. The efficiency of transduction an d expression of the transgene in severe combined immunodeficiency (SCID) mi ce-repopulating cells (SRCs) and DCs were compared between lentiviral vecto r and retroviral vectors. Induced DCs were cocultured with allogeneic or au tologous T cells to test the ability to present antigens. Results. CB CD34(+) cells transduced by lentiviral vector and expanded ex v ivo sustained stable transgene expression and multipotentiality by assessin g SRCs assay and clonogenic assay of bone marrow cells from the transplante d mice. DCs derived from these cells expressed green fluorescent protein an d surface markers CD1a, CD80, and HLA-DR and showed potent allo-stimulatory activity as well as nontransduced DCs did. On the other hand, we did not d etect transgene expression in SRCs and DCs transduced by retroviral vectors . Conclusion. Gene-modified DCs derived from ex vivo expanded CB CD34(+) cell s transduced by lentiviral vector will be useful in future immunotherapy pr otocols. (C) 2001 International Society for Experimental Hematology. Publis hed by Elsevier Science Inc.