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Generation and characterization of an immunogenic dendritic cell population.
- 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.
- Subjects :
- Animals
Antigen-Antibody Reactions
CD4-Positive T-Lymphocytes immunology
CD40 Antigens chemistry
CD40 Antigens immunology
Cell Culture Techniques
Cells, Cultured
Cytokines metabolism
Dendritic Cells cytology
Female
Flow Cytometry instrumentation
Flow Cytometry methods
Humans
Immunization methods
Immunomagnetic Separation
Immunophenotyping
Ligands
Lipopolysaccharides chemistry
Lipopolysaccharides immunology
Mice
Mice, Inbred BALB C
Phenotype
Plasmids genetics
T-Lymphocytes, Cytotoxic immunology
Viral Nonstructural Proteins chemistry
fms-Like Tyrosine Kinase 3 genetics
fms-Like Tyrosine Kinase 3 metabolism
Dendritic Cells immunology
Viral Nonstructural Proteins immunology
Subjects
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