Gene expression analysis of Vg4+ vs Vg5/8/9+ gd T cells isolated from human gut

The aim of this experiment was to characterize the the gene expression profile of colonic Vg4+ cells and to compare it to that of Vg5/8/9+ tissue-resident gd T cells. Colonic biopsies (11 per donor) from 4 donors with no intestinal pathologies were dissociated with Miltenyi Biotec's tumour dissociation kit and stained with anti-Vg4 and anti-Vg2/3/4 antibodies. Vg4+Vg2/3/4+ (termed Vg4+) and Vg4-Vg2/3/4- (termed Vg5/8/9+) cells were flow sorted directly into lysis buffer and mRNA was used for library preparation and pair-end (PE100) sequencing on on Illumina NextSeq 2000.