Phenotypical and functional evaluation of dendritic cells after exosomal delivery of miRNA-155
Phenotypical and functional evaluation of dendritic cells after exosomal delivery of miRNA-155
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DOI：
10.1016/j.lfs.2019.01.005
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发表时间：
2019-02-15
期刊：
影响因子：
6.1
通讯作者：
Amani, Davar
中科院分区：
文献类型：
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作者：
Asadirad, Ali;Hashemi, Seyed Mahmoud;Amani, Davar
目的：树突状细胞（DC）治疗的临床疗效有待提高。外泌体作为一种膜纳米囊泡，携带生物大分子，在细胞间串扰中发挥重要作用。在此，提出肿瘤细胞来源的外泌体可以用作将外源性miRNA-155递送到DC中的载体，用于同时递送miRNA和DC的抗原引发。主要方法：采用不同电压（0.100、0.200和0.300 kV）将外源性miRNA-155电穿孔至不同浓度的肿瘤细胞来源的exosomes中，然后用负载miRNA-155的exosomes处理DC。为了评估加载miRNA-155的外来体对DC的影响，通过ELISA测量培养物上清液中IL 12 p70、IFN-γ和IL 10的表达水平。结果：外源性miRNA-155可成功插入肿瘤细胞来源的exosomes中，并在体外培养的条件下，成功地将其插入到肿瘤细胞来源的exosomes中，成功地将其插入到肿瘤细胞来源的exosomes中。增强肿瘤细胞来源的外泌体的加载条件，以用作将miRNA-155递送到DC中的载体。对表面分子的分析显示，miRNA-155可以增加MHCII（I/A-I/E）、CD86、CD40和CD83的表达水平。ELISA分析表明，miRNA-155可以显著提高IL 12 p70、IFN-γ和IL 10的水平。意义：最后，可以说明miRNA-155可以作为树突状细胞成熟的候选物。该方法可应用于靶细胞的体外修饰研究。
Aims: The clinical efficiency of dendritic cell (DC) therapy needs to be improved. Exosomes, as membrane nano-vesicles, carry bio-macromolecules and play essential roles in intercellular crosstalk. Here, it is proposed that tumor cell-derived exosomes could function as vehicles to deliver exogenous miRNA-155 into DCs, for simultaneous miRNA delivery and antigen priming of DCs. Following optimization of the miRNA-155 delivery, the effect of exogenous miRNA-155 overexpression on DCs is evaluated.Main methods: For this purpose, exogenous miRNA-155 was electroporated with various voltages (0.100, 0.200, and 0.300 kV) into tumor cell-derived exosomes with various concentrations, and then DCs were treated with miRNA-155 loaded exosomes. To assess the effect of miRNA-155 loaded exosomes on DCs, the expression levels of IL12p70, IFN-gamma, and IL10 in culture supernatants were measured by ELISA. Then, the expression profiles of DC surface markers, including CD11C, MHCII (I/A-I/E), CD86, CD40, and CD83 were investigated by flow cytometry.Key findings: Concerning the results, exogenous miRNA-155 can be successfully inserted into tumor cell derived exosomes. Loading conditions for tumor cell-derived exosomes were enhanced for utilization as vehicles to deliver miRNA-155 into DCs. Analysis of the surface molecule revealed that miRNA-155 can increase the expression levels of MHCII (I/A-I/E), CD86, CD40, and CD83. ELISA analysis indicates that miRNA-155 can significantly increase, the levels of IL12p70, IFN-gamma, and IL10.Significance: Finally, it can be stated that miRNA-155 could be a candidate for dendritic cell maturation. This method can be applied in the modification of target cells in in vitro studies.