Different permeabilization patterns of splenocytes and thymocytes to combination of pulsed electric and magnetic field treatments

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 electro- magnetic 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 N14% increase in thymo- cytes permeabilization is achieved when electroporation is applied in combination with EMF, however splenocytes responded in a different manner – a statistically significant (P b 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.