Fig. 1
From: Simplified assay for enrichment of primed human Th17 and Tc17 lymphocytes from peripheral blood
Schematic representation of FACS plots showing strategy for sorting CD146+ Th17 and Tc17 cells. a Cells obtained after whole blood lysis were stained and sorted as CD45 + CD19-CD33-CD14-CD4 + CD45RO + CD146- (Th17 negative subset), CD45 + CD19-CD33-CD14-CD4 + CD45RO + CD146+ (Th17 positive subset), CD45 + CD19-CD33-CD14-CD8 + CD146- (Tc17 negative subset), and CD45 + CD19-CD33-CD14-CD8 + CD146+ (Tc17 positive subset). Lymphocytes were gated on forward and side scatter and doublets were removed. On gated CD45+ cells CD45RO + CD4 + CD146+ were gated for Th17 cells and CD8 + CD146+ as Tc17 cells. b Gating for CD146 was done on the basis of FMO controls. c Sorted cells were acquired to check purity and expression of CD3 antigen on sorted of populations. The cells were collected and stained with Diff-Quick cell staining kit to show lymphocyte morphology. d IL-17A and IFN-g secretion by freshly sorted Th17 and Tc17 positive and negative subset. As observed from the stimulated cells CD146 + CD4+ and CD146+ CD8+ cells were secreting IL-17A and IFN-g