PMID- 18258252
OWN - NLM
STAT- MEDLINE
DCOM- 20080828
LR  - 20220110
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 332
IP  - 1-2
DP  - 2008 Mar 20
TI  - Generation and characterization of an immunogenic dendritic cell population.
PG  - 18-30
LID - 10.1016/j.jim.2007.12.007 [doi]
AB  - 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.
FAU - Gehring, Stephan
AU  - Gehring S
AD  - The Liver Research Center, Providence, Rhode Island 02903, USA; The Warren Alpert 
      Medical School of Brown University, Providence, Rhode Island 02903, USA.
FAU - Gregory, Stephen H
AU  - Gregory SH
FAU - Wintermeyer, Philip
AU  - Wintermeyer P
FAU - San Martin, Maryann
AU  - San Martin M
FAU - Aloman, Costica
AU  - Aloman C
FAU - Wands, Jack R
AU  - Wands JR
LA  - eng
GR  - AA-02666/AA/NIAAA NIH HHS/United States
GR  - AA-08169/AA/NIAAA NIH HHS/United States
GR  - CA-35711/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20080114
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (CD40 Antigens)
RN  - 0 (Cytokines)
RN  - 0 (Ligands)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Viral Nonstructural Proteins)
RN  - EC 2.7.10.1 (FLT3 protein, human)
RN  - EC 2.7.10.1 (fms-Like Tyrosine Kinase 3)
RN  - EC 2.7.7.48 (NS-5 protein, hepatitis C virus)
SB  - IM
MH  - Animals
MH  - Antigen-Antibody Reactions
MH  - CD4-Positive T-Lymphocytes/immunology
MH  - CD40 Antigens/chemistry/immunology
MH  - Cell Culture Techniques
MH  - Cells, Cultured
MH  - Cytokines/metabolism
MH  - Dendritic Cells/cytology/*immunology
MH  - Female
MH  - Flow Cytometry/instrumentation/methods
MH  - Humans
MH  - Immunization/methods
MH  - Immunomagnetic Separation
MH  - Immunophenotyping
MH  - Ligands
MH  - Lipopolysaccharides/chemistry/immunology
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Phenotype
MH  - Plasmids/genetics
MH  - T-Lymphocytes, Cytotoxic/immunology
MH  - Viral Nonstructural Proteins/chemistry/*immunology
MH  - fms-Like Tyrosine Kinase 3/genetics/metabolism
EDAT- 2008/02/09 09:00
MHDA- 2008/08/30 09:00
CRDT- 2008/02/09 09:00
PHST- 2007/04/04 00:00 [received]
PHST- 2007/09/26 00:00 [revised]
PHST- 2007/12/05 00:00 [accepted]
PHST- 2008/02/09 09:00 [pubmed]
PHST- 2008/08/30 09:00 [medline]
PHST- 2008/02/09 09:00 [entrez]
AID - S0022-1759(07)00393-6 [pii]
AID - 10.1016/j.jim.2007.12.007 [doi]
PST - ppublish
SO  - J Immunol Methods. 2008 Mar 20;332(1-2):18-30. doi: 10.1016/j.jim.2007.12.007. 
      Epub 2008 Jan 14.