Artikel
Development of a Clinical-Grade Protocol to mature moncyte-derived dendritic cells in TH1-direction
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Veröffentlicht: | 20. März 2006 |
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Gliederung
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Purpose: Dendritic cells (DC) can initiate primary adaptive immune responses and skew T cell reactivity toward a TH1 or TH2 pattern. We improved the capacity of clinical-grade monocyte-derived dendritic cells to elaborate IL-12 with special regard of their state of maturation, different maturation stimuli and its regulation by TH1/TH2-influencing cytokines.
Methods: Monocyte-derived DC were generated in serum-free medium with GM-CSF and IL-4 for 7 days and matured for another 24 or 48 h with PGE2, TNF-α, ± IL-6 , ± IL-1 and ± Interferon-γ. Matured DC were phenotypically characterised and measured for IL-6, IL-10 and IL-12 p70 by ELISA.
Results: All investigated clinical-grade protocols for maturation of monocyte-derived DC appeared able to provide the phenotypic characteristics necessary to initiate DC maturation. DC matured with PGE2 and TNF-α only resulted in high levels of IL-10 but only low levels of IL-12 p70. Addition of IL-1 and IL-6 (Jonuleit cocktail) and Interferon-γ lead to significant higher levels of bioactive IL-12 concomitant with lower levels of IL-10. Besides addition of INF-γ resulted in more pronounced upregulation of CD83, major histocompatibility complex class I and B7 molecules by DC
Conclusion: PGE2, TNF-α, IL-1, Il-6 and IFN-γ represent a suitable cytokine cocktail for the ex vivo maturation of monocyte-derived DC that can be used as cellular vaccines to promote a potent type 1 immune response.
Key Words: Dendritic cells, Maturation, Adjuvants, Cytokines, Immunotherapy