Efficient ex vivo generation of dendritic cells from CD14⁺ blood monocytes in the presence of human serum albumin for use in clinical vaccine trials

Abstract

Summary. Dendritic cells (DC) with the potential to induce anti-tumour immunity represent one of the promising candidates for cancer vaccines. Efficiency of ex vivo DC generation depends on culture conditions, especially protein components in the plasma or serum used. Using human serum albumin (HSA), we devised a constant and reproducible culture method for DC generation from peripheral blood CD141 cells. The number of DC obtained with 2% HSA-supplemented cultures containing granulocyte-macrophage colony-stimulating factor and interleukin 4 were consistently higher than in cultures with various concentrations of autologous plasma or serum. The concentrations and time points tested for plasma or serum considerably affected the number of DC recovered. DC prepared with HSA acquired the ability to uptake dextran, and expressed high levels of major histocompatibility (MHC) and co-stimulatory molecules similar to DC cultured with autologous plasma or serum. Although DC cultured with autologous plasma or serum consisted of CD1a 1 and CD1a 2 populations, DC differentiated in the presence of HSA expressed CD1a. DC obtained with HSA primed and induced immunogenic peptide-specific cytotoxic T lymphocytes against a tumour rejection antigen, HER2. These findings suggest that our method for preparation of DC with HSA should prove valuable in DC generation for immunotherapy.

In vitro generation of cytotoxic T lymphocyte response using dendritic cell immunotherapy in osteosarcoma

“…As expected, an increased dextran uptake by iDCs (day 6 culture) was observed compared with the day 8 mDCs; however, the increase was not statistically significant. Various studies have indicated that the uptake of dextran may be a measure of immune response (23)(24)(25). The present results also support earlier studies that suggest that increased antigen uptake is associated with IL-12 and IFN-γ secretion, and CTL response (17,26,27).…”

Phenotype and Function of Murine Peritoneal Cavity Macrophage Derived-Dendritic Cells.

“…Several groups have succeeded in generating large numbers of functional DC from their proliferating progenitors located in bone marrow, spleen or blood in the presence of appropriate cytokines [1,4,6,8,14,16,18,22,23,30]. Other studies also suggest that monocytes can differentiate into macrophages or DC [9,19,34,40].…”

“…However, the ability of GM-CSF alone has been shown to be very limited in obtaining large numbers of cells [16]. Among the cytokines most commonly used to generate myeloid DC, GM-CSF plays a critical role and interleukin 4 (IL-4) has been shown to facilitate the differentiation of the macrophages into CD14-/CD1a+ DC lineage [1]. In all these studies, macrophage derived DC have not only efficiently stimulated lymphocytes in mixed lymphocyte reaction, but also primed T cells in vitro and in vivo, in an antigen-specific manner.…”

Uptake of Apoptotic K562 Leukaemia Cells by Immature Dendritic Cells is Greatly Facilitated by Serum

“…This difference was augmented after maturation for 2 days. In contrast, Araki et al [23] found that DCs generated in RPMI-1640 supplemented with 2% HSA had similar expression to those generated with 1% AS. Finally, Tarte et al [20] found higher expression of relevant markers on DCs generated under serum-free conditions than DCs generated with the addition of AS.…”