Journal: JCI Insight
Article Title: Nectin-4 reduces T cell effector function and is a therapeutic target in pancreatic cancer
doi: 10.1172/jci.insight.194290
Figure Lengend Snippet: ( A ) IFN-γ and TNF-α production by peripheral T cells from patients with PDAC after in vitro stimulation with anti-CD3 and anti-CD28 in the presence of plate-bound (pb, n = 8) or soluble (s, n = 4) Nectin-4. Each point represents data from 1 patient. Data are shown as mean ± SD. Unpaired 2-tailed t tests with Welch’s correction. * P < 0.05; ** P < 0.001; *** P < 0.001; **** P < 0.0001. ( B ) Representative expression level of PVRL4 by qPCR (bars indicate mean of technical duplicates). ( C ) Nectin-4 expression in PDAC PDOs by Western blot. ( D ) Dose response curves from PDAC PDOs treated with FOLFIRINOX, gemcitabine plus paclitaxel (Gem/Pac), or enfortumab vedotin. The relative viability in percentage at a given drug concentration of 2 independent biological replicates is shown. ( E ) Z scores generated from relative AUC from dose response curves from PDAC PDOs either treated with FOLFIRINOX, Gem/Pac, or enfortumab vedotin. ( F ) Representative images of 2 PDAC PDOs either treated with the standard regimen FOLFIRINOX, Gem/Pac, or enfortumab vedotin. PDOs were stained with caspase-3 dye profiling apoptosis (green) and imaged after 3 days. Scale bar: 50 μm.
Article Snippet: One day prior seeding, 96-well plates (U-bottom, Greiner Bio-One) were coated overnight with 10 μg/mL anti-CD3 (BioLegend, RRID:AB_11146991), 10 μg/mL anti-CD28 (BioLegend, RRID:AB_11148949) together with 20 μg/mL recombinant Nectin-4 protein (R&D Systems).
Techniques: In Vitro, Expressing, Western Blot, Concentration Assay, Generated, Staining