Evaluation of 3 clinical dendritic cell maturation protocols containing lipopolysaccharide and interferon-gamma.
Evaluation of 3 clinical dendritic cell maturation protocols containing lipopolysaccharide and interferon-gamma.
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DOI：
10.1097/cji.0b013e31819e1773
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发表时间：
2009-05
期刊：
影响因子：
--
通讯作者：
Stroncek DF
中科院分区：
文献类型：
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作者：
Han TH;Jin P;Ren J;Slezak S;Marincola FM;Stroncek DF
树突状细胞（DC）是癌症疫苗的重要佐剂。未成熟树突状细胞（iDCs）通常是由IL-4和GM-CSF刺激外周血单核细胞产生的。在许多应用中，dc是用细胞因子或炎症信号治疗以产生成熟的dc （mdc）。未成熟的dc通常用LPS和IFN-γ体外治疗，以产生成熟的dc用于临床治疗。本研究的目的是确定是否可以通过添加另外两种DC成熟剂IL-1β和TNF-α来改善LPS加IFN-γ的DC成熟鸡尾酒。采集6例健康人外周血单个核细胞（PBMCs）。通过洗脱从PBMC浓缩液中分离单核细胞，并与GM-CSF和IL-4孵育3天以产生iDCs。每个受试者的未成熟dc分成3个，用LPS + IFN-γ孵育24小时；LPS, IFN-γ + IL-1β；或LPS， IFN-γ， IL-1β和TNF-α产生mDCs。通过流式细胞术检测共刺激物和抗原呈递分子（CD80、CD83、CD86和HLA-DR）的表达，ELISA检测细胞因子（IL-12p70和IL-10）的产生，以及利用寡核苷酸芯片检测全局基因表达来比较DCs。在共刺激分子、HLA-DR和CCR7的表达以及IL-12p70的产生方面，3种鸡尾酒产生的mdc没有差异。全球基因表达分析发现，在3组不同的mDCs中，有9576个基因的表达存在差异，但只有13个基因的表达存在差异。在LPS和IFN-γ中添加IL-1β和TNF-α以产生mdc没有益处。
Dendritic cells (DC) are important adjuvants for cancer vaccines. Immature dendritic cells (iDCs) are often produced by the stimulation of peripheral blood monocytes with IL-4 and GM-CSF. For many applications iDCs are treatment with cytokines or inflammatory signals to produce mature DCs (mDCs). Immature DCs are often treated ex-vivo with LPS and IFN-γ to produce mature DCs for clinical therapy. The purpose of this study was to determine if the DC maturation cocktail LPS plus IFN-γ could be improved by the addition of two other DC maturation agents IL-1β and TNF-α. Peripheral blood mononuclear cells (PBMCs) were collected from 6 healthy subjects. Monocytes were isolated from the PBMC concentrates by elutriation and were incubated for 3 days with GM-CSF and IL-4 to produce iDCs. Immature DCs from each subject were divided into 3 and were incubated for 24 hours with LPS plus IFN-γ; LPS, IFN-γ plus IL-1β; or LPS, IFN-γ, IL-1β plus TNF-α to produce mDCs. The DCs were compared by measuring the expression of co-stimulator and antigen presenting molecules (CD80, CD83, CD86, and HLA-DR) by flow cytometry, cytokine production (IL-12p70 and IL-10) by ELISA and global gene expression using an oligonucleotide microarray. There were no differences in the expression of co-stimulatory molecules, HLA-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 9,576 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-1β and TNF-α to LPS and IFN-γ to order to produce mDCs.