BMS-1001

BMS-1001 lindert die hemmende Wirkung des löslichen PD-L1 auf die T-Zell-Rezeptor-vermittelte Aktivierung von T-Lymphozyten. Diese Verbindung ist wirksam bei der Abschwächung der hemmenden Wirkung des zelloberflächenassoziierten PD-L1.[1]

• Zelllinien

  PD-L1⁺ aAPC/CHO-K1 cells (aAPCs), PD-1 Effector Cells (ECs)

• Konzentrationen

  0.12 μM, 0.3 μM, 1.2 μM, 3 μM

• Inkubationszeit

  24 h

• Methode

  For the evaluation of the BMS-1001 impact on T cell inhibition by soluble PD-L1, the ECs are stimulated with the anti-CD3 antibody in the presence of the recombinant human sPD-L1. For this, the 96-well white flat bottom plates are coated overnight at 4°C with 5 μg/ml of the anti-CD3 antibody or the isotype control solution in PBS. The antibody solution is removed and the plates are washed 3 times with PBS and dried. sPD-L1 (aa 18–134) is diluted in PBS supplemented with the penicillin/streptomycin solution (100 U/ml final concentration each) in the presence of this compound or a corresponding volume of DMSO. Then, 15 μl of the solution is added to each well of the antibodycoated plate. ECs are centrifuged and diluted to 50 000 per ml, and 60 μl of the cell solution is added to each well. The final concentration of sPD-L1 was 10 μg/ml (0.6 μM). The final concentrations of this chemical are: 0.12, 0.3, 1.2 and 3 μM, giving the following BMS:sPD-L1 molar ratios: 1:5, 1:2, 2:1 and 5:1. The cells are cultured for 24 h and the luciferase activity assay is performed using the Bio-Glo Luciferase Assay System.