title: Evaluation of 3 clinical dendritic cell maturation protocols containing lipopolysaccharide and interferon-gamma.
abstract: Dendritic cells (DCs) are important adjuvants for cancer vaccines. Immature dendritic cells (iDCs) are often produced by the stimulation of peripheral blood monocytes with interleukin (IL)-4 and granulocyte macrophage-colony stimulating factor. For many applications iDCs are treated with cytokines or inflammatory signals to produce mature DCs (mDCs). iDCs are often treated ex vivo with lipopolysaccharide (LPS) and interferon (IFN)-gamma to produce mDCs for clinical therapy. The purpose of this study was to determine if the DC maturation cocktail LPS plus IFN-gamma could be improved by the addition of 2 other DC maturation agents IL-1beta and tumor necrosis factor (TNF)-alpha. Peripheral blood mononuclear cells were collected from 6 healthy subjects. Monocytes were isolated from the peripheral blood mononuclear cell concentrates by elutriation and were incubated for 3 days with granulocyte macrophage-colony stimulating factor and IL-4 to produce iDCs. iDCs from each subject were divided into 3 and were incubated for 24 hours with LPS plus IFN-gamma; LPS, IFN-gamma, plus IL-1beta; or LPS, IFN-gamma, IL-1beta, plus TNF-alpha to produce mDCs. The DCs were compared by measuring the expression of costimulator and antigen presenting molecules (CD80, CD83, CD86, and human leukocyte antigen-DR) by flow cytometry, cytokine production (IL-12p70 and IL-10) by enzyme-linked immunosorbent assay and global gene expression using an oligonucleotide microarray. There were no differences in the expression of costimulatory molecules, human leukocyte antigen-DR and CCR7 and production of IL-12p70 among the mDCs produced with the 3 cocktails. Global gene expression analysis found that the expression of 9576 genes differed between the iDCs and mDCs, but the expression of only 13 differed among the 3 different groups of mDCs. There was no benefit of adding IL-1beta and TNF-alpha to LPS and IFN-gamma to produce mDCs.
authors.affiliation: Department of Transfusion Medicine, Clinical Center, National Institutes of Health, 10 Center Drive, Bethesda, MD 20892, USA.
authors.forename: Tae Hee
authors.initials: TH
authors.lastname: Han
authors.forename: Ping
authors.initials: P
authors.lastname: Jin
authors.forename: Jiaqiang
authors.initials: J
authors.lastname: Ren
authors.forename: Stefanie
authors.initials: S
authors.lastname: Slezak
authors.forename: Francesco M
authors.initials: FM
authors.lastname: Marincola
authors.forename: David F
authors.initials: DF
authors.lastname: Stroncek