PMID- 20003055
OWN - NLM
STAT- MEDLINE
DCOM- 20100505
LR  - 20171116
IS  - 1537-2995 (Electronic)
IS  - 0041-1132 (Linking)
VI  - 50
IP  - 4
DP  - 2010 Apr
TI  - Efficient generation of clinical-grade genetically modified dendritic cells for 
      presentation of multiple tumor-associated proteins.
PG  - 831-42
LID - 10.1111/j.1537-2995.2009.02519.x [doi]
AB  - BACKGROUND: Dendritic cells (DCs) play a central role in the initiation and 
      regulation of immune responses. DCs for clinical applications can be generated 
      with high yield from leukapheresis products. Using adenoviral transduction we 
      genetically modified human DCs to produce and present melanoma-associated 
      antigens. Coexpression of green fluorescent protein and epitope tags were used to 
      monitor genetic modification. Generation, genetic modification, and 
      cryoconservation of gene modified human DCs on a clinical scale in a closed 
      system is feasible. STUDY DESIGN AND METHODS: CD14-positive monomuclear cells 
      were isolated from leukapheresis products of HLA-A* 0201 positive voluntary blood 
      donors using immunomagnetic beads. Selected cells were cultivated for 7 days. 
      Adenovirus transduction was optimal on Day 4. Maturation was induced on Day 5. 
      Mature DC were aliquoted and cryoconserved on Day 7. Quality control was 
      performed using flow cytometry, expression profiling, and functional assays 
      (ELISPOT, CBA). RESULTS: We were able to generate sufficient genetically modified 
      mature DCs in serum-free cultures that could be stored by cryopreservation. The 
      use of a closed system facilitated development of methods for standardized 
      production of clinically applicable genetically modified DCs. The adenoviral 
      transduction system allowed simultaneous and flexible expression of 
      tumor-associated antigens for prolonged presentation of multiple epitopes. 
      CONCLUSION: The feasibility of a closed-bag system for the cultivation of 
      genetically modified human DCs is shown. The immature DCs were genetically 
      modified by recombinant replication-deficient adenoviruses to express multiple 
      epitopes of tumor-associated proteins and then differentiated to mature 
      antigen-presenting DCs.
FAU - Garritsen, Henk S P
AU  - Garritsen HS
AD  - Institute for Clinical Transfusion Medicine, Braunschweig, Germany.
FAU - Macke, Lars
AU  - Macke L
FAU - Meyring, Wilhelm
AU  - Meyring W
FAU - Hannig, Horst
AU  - Hannig H
FAU - Pagelow, Ute
AU  - Pagelow U
FAU - Wormann, Bernhard
AU  - Wormann B
FAU - Geffers, Robert
AU  - Geffers R
FAU - Dittmar, Kurt E J
AU  - Dittmar KE
FAU - Lindenmaier, Werner
AU  - Lindenmaier W
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20091210
PL  - United States
TA  - Transfusion
JT  - Transfusion
JID - 0417360
RN  - 0 (Antigens, CD)
RN  - 0 (Antigens, CD19)
RN  - 0 (HLA-A Antigens)
RN  - 0 (HLA-A*02:01 antigen)
RN  - 0 (HLA-A2 Antigen)
RN  - 0 (Lipopolysaccharide Receptors)
RN  - 0 (Neoplasm Proteins)
SB  - IM
MH  - Animals
MH  - Antigens, CD/genetics/immunology
MH  - Antigens, CD19/immunology
MH  - Cell Survival/immunology
MH  - Dendritic Cells/*cytology/immunology/*physiology
MH  - HLA-A Antigens/genetics/*immunology
MH  - HLA-A2 Antigen
MH  - Histocompatibility Testing/methods
MH  - Humans
MH  - Lipopolysaccharide Receptors/genetics/immunology
MH  - Lymphocyte Activation/immunology
MH  - Neoplasm Proteins/*analysis
MH  - Organisms, Genetically Modified/*physiology
MH  - Polymerase Chain Reaction
MH  - T-Lymphocytes/immunology
MH  - Vaccination/methods
EDAT- 2009/12/17 06:00
MHDA- 2010/05/06 06:00
CRDT- 2009/12/17 06:00
PHST- 2009/12/17 06:00 [entrez]
PHST- 2009/12/17 06:00 [pubmed]
PHST- 2010/05/06 06:00 [medline]
AID - TRF2519 [pii]
AID - 10.1111/j.1537-2995.2009.02519.x [doi]
PST - ppublish
SO  - Transfusion. 2010 Apr;50(4):831-42. doi: 10.1111/j.1537-2995.2009.02519.x. Epub 
      2009 Dec 10.