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Generation and characterization of an immunogenic dendritic cell population.

Authors :
Gehring S
Gregory SH
Wintermeyer P
San Martin M
Aloman C
Wands JR
Source :
Journal of immunological methods [J Immunol Methods] 2008 Mar 20; Vol. 332 (1-2), pp. 18-30. Date of Electronic Publication: 2008 Jan 14.
Publication Year :
2008

Abstract

Dendritic cells (DCs) capture, internalize and process antigens leading to the induction of antigen-specific immune responses. The aim of this study was to develop, implement and characterize an efficient approach for DC-based immunization. Dendritic cells were expanded in vivo by hydrodynamic delivery of a human flt3 ligand expression plasmid. Splenic DCs were isolated and purified with magnetic beads linked to hepatitis C virus (HCV) nonstructural protein-5 (NS5), anti-CD40 and/or LPS. The DCs that contained beads were purified by passage over a magnetic column and subsequently phenotyped. Enrichment resulted in a population consisting of 80% CD11c(+) cells. Uptake of uncoated microparticles promoted DC maturation and the expression of CD80, CD86, and MHC-II molecules; beads coated with LPS and anti-CD40 further increased the expression of these co-stimulatory molecules, as well as the secretion of IL-12. Mice immunized subcutaneously with DCs containing beads coated with HCV NS5 protein, anti-CD40 and LPS exhibited significant antigen-specific, increases in IFN-gamma-producing CD4(+) T cells and CTL activity. This approach combines three critical elements necessary for efficient DC-based immunization that include DC enrichment, maturation and antigen targeting.

Details

Language :
English
ISSN :
0022-1759
Volume :
332
Issue :
1-2
Database :
MEDLINE
Journal :
Journal of immunological methods
Publication Type :
Academic Journal
Accession number :
18258252
Full Text :
https://doi.org/10.1016/j.jim.2007.12.007