Institute of High Magnetic Fields, Vilnius Gediminas Technical University, Vilnius, Lithuania.
State Research Institute Centre for Innovative Medicine, Department of Immunology, Vilnius, Lithuania.
Bioelectrochemistry. 2018 Aug;122:183-190. doi: 10.1016/j.bioelechem.2018.04.006. Epub 2018 Apr 10.
Genetic manipulation of T cells is frequently inefficient, however, when combined with physical methods (i.e. electroporation) a promising alliance with immunotherapy can be formed. This study presents new data on permeabilization of murine thymocytes and splenocytes as a T cell model using pulsed electric (PEF) and electromagnetic field (EMF). The 300ns, 500ns, 2μs and 100μs pulse bursts in a broad range of PEF 0-8kV/cm were applied separately and in combination with 3.3T, 0.2kV/cm EMF pulses. The permeabilization efficiency was evaluated using fluorescent dye (YO-PRO-1) and flow cytometry. It was shown that a >14% increase in thymocytes permeabilization is achieved when electroporation is applied in combination with EMF, however splenocytes responded in a different manner - a statistically significant (P<0.05) reduction in permeabilization was observed. The cytokine secretion patterns were mainly unaltered independently on the applied treatment parameters determined by secretion of IFNγ, IL-4 and IL-17 - the main cytokines of Th1, Th2 and Th17 cells. The results of this study are useful for development of pulsed power protocols for effective genetic modification of T cells.
然而，当基因操作与物理方法（如电穿孔）结合时，就可以与免疫疗法形成有前途的联盟。本研究提出了新的数据，即在使用脉冲电场（PEF）和电磁场（EMF）作为 T 细胞模型时，对鼠胸腺细胞和脾细胞进行通透化的情况。分别应用了 300ns、500ns、2μs 和 100μs 的宽范围 PEF 0-8kV/cm 的脉冲爆发，以及与 3.3T、0.2kV/cm EMF 脉冲的组合。使用荧光染料（YO-PRO-1）和流式细胞术评估了通透化效率。结果表明，当电穿孔与 EMF 联合应用时，胸腺细胞的通透化率增加了>14%，然而脾细胞的反应方式不同——观察到通透化显著降低（P<0.05）。细胞因子分泌模式主要不受应用处理参数的影响，这些参数由 IFNγ、IL-4 和 IL-17 的分泌决定，这些是 Th1、Th2 和 Th17 细胞的主要细胞因子。本研究的结果有助于开发有效的 T 细胞基因修饰的脉冲功率方案。
