BLITC reporter for monitoring of GvHD. BLITC reporter for monitoring of GvHD. H-Y specific CD4+ T cells were harvested from female ML-BLITC mice and sorted to high purity (>98%). T cells were activated and expanded for 3 days. At day of ATT, male and female (control) mice were sublethally irradiated (2 × 1.5 Gy within 4 hours) before i.v. injection of 5 × 104 (males) or 5 × 105 ML-BLITC T cells (females). GvHD was monitored daily by scoring as well as BLI for NFAT-CBR signals (mean + SEM; BLI and scoring data representative of four independent experiments; n = 3–4 mice). BLI intensities were measured by setting ROI encompassing the entire mouse trunk. A, Total body NFAT-CBR signals were plotted against GvHD scores. B, BLI images for both Rluc (left) as well as NFAT-CBR signals (right) are shown for the time of GvHD duration. Rluc signals for day 8 were not acquired due to GvHD status of affected mice. C, In a similar experiment with identical settings demonstrating similar GvHD progression, female mice were sacrificed for T-cell analysis at day 6 and male mice at days 6 (male on peak) and 16 (male off peak) after T-cell transfer. Cells were isolated from spleen, liver, and mesenteric lymph nodes and analyzed for CD25 expression (left). IFNy serum levels were measured via ELISA. Graphs display mean + SEM (n = 5 mice). Martin Szyska et al. Cancer Immunol Res 2018;6:110-120 ©2018 by American Association for Cancer Research