Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Life Sci. 2019 Feb 15;219:152-162. doi: 10.1016/j.lfs.2019.01.005. Epub 2019 Jan 6.
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.
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-γ, 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.
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-γ, and IL10.
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.
树突状细胞（DC）治疗的临床疗效有待提高。外泌体作为膜纳米囊泡，携带生物大分子，在细胞间通讯中发挥重要作用。在这里，提出肿瘤细胞衍生的外泌体可以作为载体将外源性 miRNA-155 递送到 DC 中，同时进行 miRNA 递呈和 DC 的抗原启动。在外泌体递送 miRNA-155 得到优化后，评估外源性 miRNA-155 过表达对 DC 的影响。
为此，用不同浓度的外泌体将外源性 miRNA-155 用不同的电压（0.100、0.200 和 0.300kV）电穿孔，然后用负载 miRNA-155 的外泌体处理 DC。为了评估负载 miRNA-155 的外泌体对 DC 的影响，通过 ELISA 测量培养上清液中 IL12p70、IFN-γ 和 IL10 的表达水平。然后，通过流式细胞术研究 DC 表面标志物的表达谱，包括 CD11C、MHCII（I/A-I/E）、CD86、CD40 和 CD83。
关于结果，外源性 miRNA-155 可以成功插入肿瘤细胞衍生的外泌体。负载肿瘤细胞衍生的外泌体的条件得到增强，以用作将 miRNA-155 递送到 DC 的载体。表面分子分析表明，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 可以显著增加 IL12p70、IFN-γ 和 IL10 的水平。
最后，可以说 miRNA-155 可以作为树突状细胞成熟的候选物。该方法可应用于体外研究中靶细胞的修饰。
