Journal: The Journal of Clinical Investigation
Article Title: Blockade of the immunosuppressive KIR2DL5/PVR pathway elicits potent human NK cell–mediated antitumor immunity
doi: 10.1172/JCI163620
Figure Lengend Snippet: ( A – C ) Redirected cytotoxicity of KIR2DL5 + primary NK cells against P815. ( A ) The lysis of P815 cells ( n = 4). ( B ) The degranulation (CD107a) and cytokine production (IFN-γ and TNF-γ) of KIR2DL5 + primary NK cells ( n = 4). CD56 served as negative control. ( C ) Cytokine production in the coculture supernatant of KIR2DL5 + primary NK cells with the indicated antibody-coated P815 ( n = 5). Data are represented as mean ± SEM. ( D ) Lysis of scrambled control or PVR KO A427 or Jurkat cells by KIR2DL5-transduced primary NK cells (KIR2DL5/NK) or KIR2DL5 – control NK cells (Control NK) at indicated E/T ratios. ( E ) PVR-KIR2DL5–mediated inhibitory synapse formation. Left: Representative imaging of cell conjugates acquired upon sorted KIR2DL5 + primary NK contact with control-YFP/Raji (top) or PVR-YFP/Raji (bottom), followed by staining with anti-KIR2DL5 mAbs and phalloidin. Scale bars: 10 μm. Right: Intensity quantification of F-actin, YFP, and KIR2DL5 at the immunological synapses (IS) and the cell surface away from synapses (Non-IS) from KIR2DL5 + NK cell–Control Raji ( n = 25) and KIR2DL5 + NK-PVR/Raji ( n = 35) conjugates. ( F ) Lysis of scrambled control or PVR KO A427 (top) or Jurkat (bottom) cells by sorted KIR2DL5 + primary NK cells in the presence of F8B30 or mIgG1 at indicated E/T ratios. In D and F , data are mean for duplicate measurements and representative of 3 independent experiments with 3 different donors. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, by 1-way ANOVA ( A and B ), 2-tailed paired Student’s t test ( C and E ), or multiple unpaired t test ( D and F ).
Article Snippet: Mouse cell lines used in this study were mouse fibroblast line NIH 3T3 (ATCC, CRL-1658), mouse mast cell line P815 (ATCC, TIB-64), and mouse myeloma cell line NSO (a gift from Matthew D. Scharff, Department of Cell Biology, Albert Einstein College of Medicine).
Techniques: Lysis, Negative Control, Control, Imaging, Staining
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