Generation of dendritic cell-based vaccines for cancer therapy.
Bottom Line: Dendritic cells play a major role in the generation of immunity against tumour cells.They can be grown under various culture conditions, which influence the phenotypical and functional properties of dendritic cells and thereby the consecutive immune response mainly executed by T cells.Here we discuss various conditions, which are important during generation and administration of dendritic cells to elicit a tumouricidal T cell-based immune response.
Affiliation: Klinik und Poliklinik für Dermatologie der Rheinischen Friedrich-Wilhelms-Universität Bonn, Germany.Show MeSH
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Mentions: DC can either be generated from proliferating CD34+ bone marrow precursor cells (Caux et al, 1996) – which differentiate under a variety of different cytokines including SCF, Flt3, GM–CSF, TGF-β and TNF-α – or from non-proliferating peripheral CD14+ cells (monocytes) (Sallusto and Lanzavecchia, 1994). Usually, CD34+ precursors mobilised by G-CSF are isolated by leukapheresis to obtain high numbers of peripheral cells for therapeutical purposes. These cells seem to be more efficient in the activation of tumour-specific CTLs than CD14+ derived DC (Mortarini et al, 1997). CD34+ cells expand 10–30-fold. Yields of 5×106 cells per leukapheresis are typically obtained. In contrast, monocytes are abundantly present in peripheral blood and can be easily obtained by peripheral blood drawings or leukapheresis. Protocols for the generation of large amounts of monocyte-derived DC are known since 1994 (Romani et al, 1994; Sallusto and Lanzavecchia, 1994) and have been used for both experimental and therapeutical purposes. Here, leukocytes are prepared from peripheral blood using Ficoll–Hypaque density centrifugation. Monocytes are isolated by an adherence step and subsequently cultured in the presence of GM–CSF, IL-4 and 10% FCS or alternatively – under serum free conditions (Jonuleit et al, 1997; Thurner et al, 1999b) – with 1% autologous plasma for 7 days (Figure 1Figure 1
Affiliation: Klinik und Poliklinik für Dermatologie der Rheinischen Friedrich-Wilhelms-Universität Bonn, Germany.