Summit software v6

Flow cytometry was performed using the following anti-bodies: APC anti-human CD4 (OKT4), Brilliant Violet 711 anti-human CD4 (OKT4), APC anti-human CD56 (HCD56), FITC anti-human CD8a (RPA-T8), Pacific Blue anti-human CD3 (HIT3a), and PE anti-human alpha/beta-TCR (IP26) (BioLegend, San Diego, CA). Analysis and sorting were performed using MoFlow Astrios and Summit software v6.2.3 (Beckman Coulter, Miami, FL). First, live samples were plotted by forward scatter (FSC) and side scatter channels (SSC) then gated on T cells (Fig. 1). Cell aggregates were excluded by gating on T cells plotted by SSC-width versus SSC-height and FSC-width versus FSC-height. For cell purity analyses, T cells were plotted in the Pacific Blue and APC channels to show N 97% CD3+ T cell purity using the Pacific Blue anti-human CD3 antibody and N 98% CD3+CD56− purity from invariant NK T cells using APC anti-human CD56 (Fig. 1A). For cell sorting, T cells were plotted by side scatter and PE channels to gate TCRαβ+ cells (Fig. 1B). These cells were subsequently plotted by APC and FITC channels to sort CD4+, CD8+, and DN T cells.