gms | German Medical Science

56. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e. V. (DGNC)
3èmes journées françaises de Neurochirurgie (SFNC)

Deutsche Gesellschaft für Neurochirurgie e. V.
Société Française de Neurochirurgie

07. bis 11.05.2005, Strasbourg

Serum-free generation of mature dendritic cells from malignant glioma patients

Serumfreie Gewinnung reifer dendritischer Zellen von Gliompatienten

Meeting Abstract

  • Z. Özcan - Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Düsseldorf
  • corresponding author M. Rapp - Department of Neurosurgery, Heinrich-Heine-University Medical Center, Düsseldorf
  • S. Vleeschouwer - Department of Neurosurgery, Heinrich-Heine-University Medical Center, Düsseldorf
  • W. Stummer - Department of Neurosurgery, Heinrich-Heine-University Medical Center, Düsseldorf
  • P. Wernet - Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Düsseldorf
  • H.-J. Steiger - Department of Neurosurgery, Heinrich-Heine-University Medical Center, Düsseldorf
  • R. Sorg - Institute for Transplantation Diagnostics and Cell Therapeutics, Heinrich-Heine-University Medical Center, Düsseldorf
  • M. Sabel - Department of Neurosurgery, Heinrich-Heine-University Medical Center, Düsseldorf

Deutsche Gesellschaft für Neurochirurgie. Société Française de Neurochirurgie. 56. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie e.V. (DGNC), 3èmes journées françaises de Neurochirurgie (SFNC). Strasbourg, 07.-11.05.2005. Düsseldorf, Köln: German Medical Science; 2005. DocP171

Die elektronische Version dieses Artikels ist vollständig und ist verfügbar unter: http://www.egms.de/de/meetings/dgnc2005/05dgnc0439.shtml

Veröffentlicht: 4. Mai 2005

© 2005 Özcan et al.
Dieser Artikel ist ein Open Access-Artikel und steht unter den Creative Commons Lizenzbedingungen (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.de). Er darf vervielf&aauml;ltigt, verbreitet und &oauml;ffentlich zug&aauml;nglich gemacht werden, vorausgesetzt dass Autor und Quelle genannt werden.


Gliederung

Text

Objective

Immunotherapy with tumor-antigen loaded dendritic cells (DC) is a promising approach, which may allow the specific killing of tumor cells without excessive damage to normal tissues. However, it relies on ex-vivo generation of sufficient numbers of functionally competent DC, which may be impaired in patients. Here, mature monocyte-derived DC were generated from malignant glioma patients in a two-step culture protocol using either plasma-supplemented or serum-free medium.

Methods

DC generation in glioma patients and controls was compared, using GMP-quality, serum-free CellGro DC 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

In patients (n=14), frequencies of lymphocytes and CD3+ T-cells were significantly reduced, whereas the frequency of neutrophils was increased. Immunophenotypic characterization of T-cells revealed a decreased frequency of CD25+ cells in patients. Functionally, there was a slightly better mitogen-responsiveness of T-cells in patients as compared to controls. In the monocyte compartment, the only phenotypic difference observed was a lower frequency of CD80+ cells in patients. DC differentiation of monocytes revealed more homogenous generation of mature CD83+/CD14- DC for patients compared to controls in the presence of X-vivo 15 medium supplemented with autologous plasma. Residual CD14 expression was lower and expression of CD83, CD80 and CD25 was higher for patients. When serum-free CellGro DC medium was used instead, all of these differences disappeared, besides the higher frequency of CD25+ DC in patients.

Conclusions

Overall, the serum-free medium was superior to plasma-supplemented X-vivo 15 medium. Thus, only minor differences between glioma patients and controls were observed, which probably will not affect DC-based immunotherapy. In addition, a serum-free culture system is defined, which allows efficient and homogenous ex vivo generation of mature DC from glioma patients for clinical application.