Investigations of the functional states of dendritic cells under different conditioned microenvironments by Fourier transformed infrared spectroscopy.
Investigations of the functional states of dendritic cells under different conditioned microenvironments by Fourier transformed infrared spectroscopy.
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傅里叶变换红外光谱研究不同条件微环境下树突状细胞的功能状态
DOI：
10.1186/1475-925x-13-2
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发表时间：
2014-01-10
影响因子：
3.9
通讯作者：
Zeng Z
中科院分区：
文献类型：
--
作者：
Dong R;Long J;Xu X;Zhang C;Wen Z;Li L;Yao W;Zeng Z
树突状细胞是一种强大的特化抗原呈递细胞，在先天免疫和适应性免疫反应的启动和扩增中起着至关重要的作用。以树突状细胞为基础的抗癌疫苗在临床上取得了可喜的成功。但其临床应用仍面临诸多挑战，特别是如何识别其功能状态。方法从人外周血中分离经塑料粘附的CD14+单核细胞，用鸡尾酒免疫磁珠纯化至98%左右。采用传统方法诱导未成熟树突状细胞和成熟树突状细胞。将得到的树突状细胞与正常细胞和癌细胞共培养。利用傅里叶变换红外光谱（FTIR）和分子生物学方法研究了不同条件微环境下树突状细胞(包括未成熟树突状细胞（imdc）和成熟树突状细胞（mDCs）的功能状态。结果傅里叶变换红外光谱结果显示，肿瘤细胞特异性抑制树突状细胞的基因转录活性和能量状态（P< 0.05或0.01）。树突状细胞NF-κB （NF-κB）表达水平也受到肿瘤源性因子的特异性抑制（P< 0.05或0.01）。傅里叶变换红外光谱在给定波数下的吸收强度比值与NF-κB表达水平密切相关（R2:0.69和R2:0.81）。结论傅里叶变换红外光谱在给定波数下的吸收强度比值与NF-κB表达水平呈正相关，提示傅里叶变换红外光谱技术可在临床应用于树突状细胞接种时鉴定树突状细胞的功能状态。这对简化和规范基于树突状细胞的疫苗临床制备方案具有重要意义。
BackgroundDendritic cells are potent and specialized antigen presenting cells, which play a crucial role in initiating and amplifying both the innate and adaptive immune responses. The dendritic cell-based vaccination against cancer has been clinically achieved promising successes. But there are still many challenges in its clinical application, especially for how to identify the functional states.MethodsThe CD14+monocytes were isolated from human peripheral blood after plastic adherence and purified to approximately 98% with cocktail immunomagnetic beads. The immature dendritic cells and mature dendritic cells were induced by traditional protocols. The resulting dendritic cells were cocultured with normal cells and cancer cells. The functional state of dendritic cells including immature dendritic cells (imDCs) and mature dendritic cells (mDCs) under different conditioned microenvironments were investigated by Fourier transformed infrared spectroscopy (FTIR) and molecular biological methods.ResultsThe results of Fourier transformed infrared spectroscopy showed that the gene transcription activity and energy states of dendritic cells were specifically suppressed by tumor cells (P< 0.05 or 0.01). The expression levels of NF-kappa B (NF-κB) in dendritic cells were also specifically inhibited by tumor-derived factors (P< 0.05 or 0.01). Moreover, the ratios of absorption intensities of Fourier transformed infrared spectroscopy at given wave numbers were closely correlated with the expression levels of NF-κB (R2:0.69 and R2:0.81, respectively).ConclusionOur results confirmed that the ratios of absorption intensities of Fourier transformed infrared spectroscopy at given wave numbers were positively correlated with the expression levels of NF-κB, suggesting that Fourier transformed infrared spectroscopy technology could be clinically applied to identify the functional states of dendritic cell when performing dendritic cell-based vaccination. It’s significant for the simplification and standardization of dendritic cell-based vaccination clinical preparation protocols.