RNA sequencing of ex vivo human colonic mucosal Vg2/3/4+ CD103+ and CD103negative T cells

Purpose: to characterise the steady state gene expression and transcriptomic characteristic of tissue residernt human colonic mucosal gd T cells from controls undergoing routine colonoscopy. Overall design: Methods: 12 fresh pinch biopsies were obtained per patient from 4 control patients underoing screening colonoscopy and immediately dissociated using a Miltenyl tumour dissociation protocol. Cells were FACS sorted according to their surface phenotype Vg2/3/4 CD103+ or CD103neg. For the first 3 donors these were frozen and sent to Genewiz for RNA extrraction library preparation and sequencing on an Illumina HiSeq. For the final donor, RNA was purified immediately after the sort and libraries were prepared and sequenced on an Illumina HiSeq 2500 at the Francis Crick Institute in London.