Different permeabilization patterns of splenocytes and thymocytes to combination of pulsed electric and magnetic field treatments
Vitalij Novickij, Auksė Zinkevičienė, Justinas Valiulis, Jurgita Švedienė, Algimantas Paškevičius, Eglė Lastauskienė, Svetlana Markovskaja, Jurij Novickij, Irutė Girkontaitė
Index: 10.1016/j.bioelechem.2018.04.006
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Abstract
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 300 ns, 500 ns, 2 μs and 100 μs pulse bursts in a broad range of PEF 0–8 kV/cm were applied separately and in combination with 3.3 T, 0.2 kV/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.