Effect of treatment with Levamisole 1 mM on activated human T-cells

To investigate the effects of levamisole treatment on in vitro activated human T-cells, we isolated T-cells from buffy coats from 3 donors. All buffy coats were collected from adult anonymous healthy blood donors (Sanquin Blood Supply, Amsterdam, The Netherlands) who provided written informed consent for usage of the remainder of their blood donation in research. Untouched T cells were isolated from PBMCs by negative Magnetic-activated cell sorting (MACS) using the human pan T cell isolation kit (Miltenyi Biotec) according to the manufacturer’s instructions. Purity of the T cell fraction was assessed by flowcytometry for CD3 (>98% for all donors). To investigate the effects of Levamisole treatment on the T-cell immune response we performed bulk RNA sequencing on 24-hour and 48-hour aCD3/aCD28 dynabeads activated T-cells (Gibco, Thermo Fisher Scientific). Levamisole samples were treated with levamisole 1 mM while control samples were treated with the appropriate solvent controls. 3 separate donors were included in this experiment.