PMID- 12547597
OWN - NLM
STAT- MEDLINE
DCOM- 20031030
LR  - 20190728
IS  - 0264-410X (Print)
IS  - 0264-410X (Linking)
VI  - 21
IP  - 9-10
DP  - 2003 Feb 14
TI  - Generation of antigen-loaded dendritic cells in a serum-free medium using 
      different cytokine combinations.
PG  - 877-82
AB  - Dendritic cells (DCs) are antigen-presenting cells that play a critical role in 
      the induction of cytotoxic T-lymphocytes. An optimal method for the generation of 
      DC for clinical use remains to be established. The aim of our study was to find 
      an optimal cytokine combination for DC generation from peripheral blood stem 
      cells (PBSC) and peripheral blood mononuclear cells (PBMC) in serum-free 
      conditions. Serial immunophenotyping enabled us to observe changes in DC content 
      during the culture as well as the development of maturation and activation 
      markers. As a source for DC culture, we used PBSC from patients with multiple 
      myeloma after stem cell mobilization using cyclophosphamide and G-CSF, or PBMC 
      from healthy donors without mobilization. The cells were cultured in a serum-free 
      medium with different cytokine combinations including GM-CSF, TNF-alpha, Flt-3, 
      CD40L, IFN-gamma, IL-1alpha, IL-6, PGE1, and IL-4. The cell cultures were 
      evaluated by immunophenotyping. For PBMC, interleukin-12 assay was performed. For 
      PBSC, the yield of DC as determined by CD83+ cell count ranged from 0. 6 x 10(5) 
      to 30.1 x 10(4) (mean: 9.4 x 10(4)) of DC generated per 1 x 10(6) of initially 
      plated nucleated cells from apheresis. This yield corresponded to (0.3-19.1) x 
      10(5) (mean: 4.3 x 10(5)) per 1 x 10(6) of CD34+ cells in the apheresis products. 
      For PBMC, the yield was (0.4-24.8) x 10(4) (mean: 2.4 x 10(4)) of DC generated 
      per 1 x 10(6) of initially plated mononuclear cells from venous blood. The 
      cultured cells expressed the mature immunophenotype. No significant differences 
      in cell yield or immunophenotype were detected when comparing different cytokine 
      combinations.
FAU - Buchler, Tomas
AU  - Buchler T
AD  - Department of Clinical Hematology, Masaryk University Hospital, Brno, Czech 
      Republic. tbuchler@fnbrno.cz
FAU - Hajek, Roman
AU  - Hajek R
FAU - Bourkova, Lida
AU  - Bourkova L
FAU - Kovarova, Lucie
AU  - Kovarova L
FAU - Musilova, Romana
AU  - Musilova R
FAU - Bulikova, Alena
AU  - Bulikova A
FAU - Doubek, Michal
AU  - Doubek M
FAU - Svobodnik, Adam
AU  - Svobodnik A
FAU - Mareschova, Iveta
AU  - Mareschova I
FAU - Vanova, Pavlina
AU  - Vanova P
FAU - Tuzova, Eva
AU  - Tuzova E
FAU - Vidlakova, Petra
AU  - Vidlakova P
FAU - Vorlicek, Jiri
AU  - Vorlicek J
FAU - Penka, Miroslav
AU  - Penka M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Vaccine
JT  - Vaccine
JID - 8406899
RN  - 0 (Antigens)
RN  - 0 (Cancer Vaccines)
RN  - 0 (Culture Media, Serum-Free)
RN  - 0 (Cytokines)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 187348-17-0 (Interleukin-12)
RN  - 207137-56-2 (Interleukin-4)
RN  - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor)
SB  - IM
MH  - Antigens/*administration & dosage
MH  - Cancer Vaccines/administration & dosage
MH  - Cell Differentiation
MH  - Cells, Cultured
MH  - Culture Media, Serum-Free
MH  - Cytokines/*administration & dosage
MH  - Dendritic Cells/cytology/*immunology
MH  - Granulocyte-Macrophage Colony-Stimulating Factor/administration & dosage
MH  - Humans
MH  - Immunophenotyping
MH  - Immunotherapy
MH  - Interleukin-12/biosynthesis
MH  - Interleukin-4/administration & dosage
MH  - Recombinant Proteins/administration & dosage
MH  - Tumor Necrosis Factor-alpha/administration & dosage
EDAT- 2003/01/28 04:00
MHDA- 2003/10/31 05:00
CRDT- 2003/01/28 04:00
PHST- 2003/01/28 04:00 [pubmed]
PHST- 2003/10/31 05:00 [medline]
PHST- 2003/01/28 04:00 [entrez]
AID - S0264410X02005352 [pii]
AID - 10.1016/s0264-410x(02)00535-2 [doi]
PST - ppublish
SO  - Vaccine. 2003 Feb 14;21(9-10):877-82. doi: 10.1016/s0264-410x(02)00535-2.