Artikel
Ex-Vivo dendritic cell generation from CD14+ monocytes under serum-free conditions for patients with malignant glioma
Serumfreie Gewinnung dendritischer Zellen ex vivo aus CD14+ Monocyten für Gliompatienten
Suche in Medline nach
Autoren
-
Z. Özcan
- Institut für Transplantations-Diagnostik und Zelltherapeutik, Heinrich-Heine-Universität, Düsseldorf
-
M. Gorol
- Neurochirurgische Klinik, Heinrich-Heine-Universität, Düsseldorf
-
R. V. Sorg
- Institut für Transplantations-Diagnostik und Zelltherapeutik, Heinrich-Heine-Universität, Düsseldorf
-
Michael Christoph Sabel
- Neurochirurgische Klinik, Heinrich-Heine-Universität, Düsseldorf
| Veröffentlicht: | 23. April 2004 |
|---|
Gliederung
Text
Objective
Vaccination therapy with autologous, tumor-antigen pulsed dendritic cells (DC) is a promising immunotherapeutic approach. It requires ex-vivo generation of adequate numbers of mature DC. Most protocols for ex-vivo DC generation for clinical application rely on the use of autologous serum/plasma. Serum/plasma from glioma patients, however, may not be suitable, due to factors produced by tumor cells (e.g. cytokines like TGFβ) or medication (e.g. steroids). Therefore, a serum/plasma-free culture system would be preferable.
Methods
DC generation in patients with malignant gliomas (7x GBM IV, 1x AA III ) and controls (n=13) was compared, using GMP-quality, serum-free CellGroDC medium and plasma-supplemented X-Vivo 15 medium. Monocytes were enriched immunomagnetically to over 96% CD14+ purity and cultured in the presence of GM-CSF and IL-4 for the first 6 days, followed by an additional 3-day culture period in the presence of GM-CSF, IL-4 and TNFα, to induce DC maturation. Resulting DC were characterized by flow cytometry.
Results
There were no differences between patients and controls in the incidence of CD14- (2.6±1.2% vs. 0.7±0.2%) and CD83+ (66.2±8.0% vs. 65.8±6.8%) mature DC, which expressed the CD40, CD80, CD86 and HLA-DR antigens. Yield of DC calculated from monocyte input was comparable (24.1±3.3% vs. 23.9±4.4%). The only difference observed was a higher incidence of CD25+ DC (9.2±3.0% vs. 2.0±0.2%; p=0.007) for patients. Compared to the standard procedure using autologous plasma, for patients and controls, CellGroDC medium was superior in ex-vivo generation of mature CD14-/CD83+ DC.
Conclusions
CellGroDC medium allows efficient DC generation from glioma patients under serum/plasma-free conditions and should be suitable for ex-vivo generation of DC for clinical application.
