Human monocyte isolation methods influence cytokine production from in vitro generated dendritic cells
Human monocyte isolation methods influence cytokine production from in vitro generated dendritic cells
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DOI：
10.1111/j.1365-2567.2004.02076.x
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发表时间：
2005-02-01
期刊：
影响因子：
6.4
通讯作者：
Rowbottom, AW
中科院分区：
文献类型：
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作者：
Elkord, E;Williams, PE;Rowbottom, AW
人们对外周血单核细胞体外生成树突状细胞（DC）越来越感兴趣，但是选择分离CD14（+）单核细胞用于随后DC生成的方法的效果文献很少。分离单核细胞的方法可能通过影响DC表达共刺激分子（CD80/86）、成熟标记物（CD83）和/或产生重要免疫调节细胞因子的能力，对DC的后续功能产生影响。在这项研究中，我们发现抗cd14包被的微球对单核细胞的阳性选择抑制了脂多糖（LPS）诱导的白细胞介素(IL)-12、IL-10和肿瘤坏死因子- α (tnf - α)的产生。然而，当DC从塑料粘附分离的单核细胞中生长时，LPS诱导产生更高水平的这些细胞因子。经干扰素- γ (ifn - γ)酶联免疫吸附斑点形成细胞试验（ELISPOT）、细胞毒性试验、主要组织相容性复合体(MHC)-肽四聚体复合体和辅助性T细胞1/辅助性T细胞2 （Th1/Th2）细胞因子生成试验证实，来自粘附分离单核细胞的DC诱导了流感基质蛋白特异性Tc1亚群的强效细胞毒性T淋巴细胞的发育。
There is growing interest in the in vitro generation of dendritic cells (DC) from peripheral blood monocytes, but the effect of the method chosen to isolate CD14(+) monocytes for subsequent DC generation is poorly documented. The method used to isolate monocytes may have an impact on the subsequent function of DC by affecting their ability to express costimulatory molecules (CD80/86), maturation marker (CD83) and/or to produce important immunomodulatory cytokines. In this study, we show that the positive selection of monocytes by anti-CD14-coated microbeads inhibits the lipopolysaccharide (LPS)-induced production of interleukin (IL)-12, IL-10 and tumour necrosis factor-alpha (TNF-alpha) from human DC. However, when DC were grown from monocytes isolated by plastic adherence, LPS induced the production of much higher levels of these cytokines. DC derived from adherence-isolated monocytes induced the development of potent cytotoxic T lymphocytes of the Tc1 subset specific for influenza matrix protein, as confirmed by interferon-gamma (IFN-gamma) enzyme-linked immunosorbent spot-forming cell assay (ELISPOT), cytotoxicity assay, major histocompatibility complex (MHC)-peptide tetrameric complexes and T helper 1/T helper 2 (Th1/Th2) cytokine production assays.