PD-1, but not PD-L1, expressed by the BDC2

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PD-1, but not PD-L1, expressed by the BDC2 PD-1, but not PD-L1, expressed by the BDC2.5 T cell is required to limit antigen-specific proliferation, CXCR3 expression, and infiltration of the pancreas. PD-1, but not PD-L1, expressed by the BDC2.5 T cell is required to limit antigen-specific proliferation, CXCR3 expression, and infiltration of the pancreas. A: The absolute number of WT or PD-1 KO BDC2.5 T cells was measured 7 days posttransfer using the low-transfer model. Data are representative of six experiments containing spleen, pLN, and pancreas, five of which also contained inguinal LNs, with three to four mice for each genotype. B: The absolute number of WT or PD-L1 KO BDC2.5 T cells was determined 8 days posttransfer. Data are representative of three experiments containing spleen, pLN, and pancreas, two of which also contained inguinal LNs, with three to four mice per genotype. C: Flow cytometry histograms showing CXCR3 expression of WT or PD-1 KO BDC2.5 T cells 7 days posttransfer. D: Frequency of CXCR3+ WT or PD-1 KO BDC2.5 T cells 7 days posttransfer. E: Histological analysis of pancreas from mice 7–8 days posttransfer of WT or PD-1 KO BDC2.5 T cells. Gray is insulin, red is Thy1.1+ BDC2.5 T cells, green is CD3+ T cells, and DAPI marking nuclei is blue. Single color images showing BDC2.5 T cells in red are to the right of the overlaid image containing all four colors. White lines in the red single color image show the outline of the islet body containing the insulin-producing cells. BDC2.5 T cells are marked with yellow arrows in the WT group to aid in the visualization of the cells. Scale bar = 100 μm. F: Insulitis was scored blinded from mice 7–8 days posttransfer of WT or PD-1 KO BDC2.5 T cells. Thirty islets from WT and sixty-four islets from PD-1 KO transfers were scored blinded. Insulitis scores: 1, peri-insulitis or <25% infiltrated; 2, 25–50% infiltrated; and 3, >50% infiltrated. E and F: Two independent experiments with four mice per group. ingLN, inguinal LN; max, maximum; pancLN, pancreatic LN. ns, not significant, P > 0.05. *Significant, P = 0.01–0.05. **Very significant, P = 0.001–0.01. ***Extremely significant, P < 0.001. Kristen E. Pauken et al. Diabetes 2013;62:2859-2869 ©2013 by American Diabetes Association