Fig. 5
T cells enhance CXCL10-induced melanocyte death by induction of adaptive immunity. a CXCL10-induced death of vitiligo melanocytes in presence or absence of patients own autologous T cells. As above, IFNγ-pretreated melanocytes were exposed to CXCL10 in presence or absence of CXCR3 antagonist AS612568 (2 μM). The next day, media was replaced and 3 days later patient’s own CD3+ T cell were sorted and added to the melanocytes (n = 3) prior to initiation of IncuCyte. b At the end of the experiment (~48 h), supernatant was collected, remaining cells trypsinised and cytospin sections prepared for immunofluorescence detection of co-stimulatory (CD40, CD80, HLA-DR) and adhesion (ICAM-1) molecules on melanocytes. c T cell-induced potentiation of melanocyte death in IFNγ-pretreated melanocytes (compared to untreated melanocytes) was associated with a parallel increase in the number of CD3+ T cells which was supported by increased expression of Ki67+cells in the same cytospin sections d. Results are shown as individual dot plots with a line at mean ± SEM. e T cell proliferation was quantified by flow analysis measuring the percentage of CD3+CSFE+ cells undergoing 0, 1, 2 or 3+divisions (n = 3). Labelled cells at time zero was used as a negative reference, unstimulated cells left in culture for 72 h before labelling as a control and cells stimulated with PHA for 72 h (Phytohemagglutinin, 5 μg/ml) as a positive control
