Pokeweed pwd

Roswell Park Memorial Institute (RPMI) medium (Invitrogen) was used for the cell cultures. The LPS, from Escherichia coli O111:B4, was obtained from Sigma-Aldrich. Carboxyfluorescein succinimidyl ester (CFSE) for the proliferation assays was purchased from Thermo Fisher. The CF-adapted ST395 P. aeruginosa clone was isolated from a patient who had been colonized during 13 years. LPS was obtained as previously described [23] . Colistimethate Sodium (polymyxin E) was purchased from Genéricos Españoles (GES) and dissolved in RPMI medium. The lymphocyte stimulus pokeweed (PWD) was purchased from Sigma-Aldrich. To inhibit PD-L1/PD-1 interaction, a fully human IgG4 (S228P) anti-PD-1 receptor-blocking monoclonal antibody was used (Bristol-Myers Squibb). All the reagents used for the cell cultures were endotoxin-free, as assayed with the Limulus amebocyte lysate test (Cambrex).

The carboxyfluorescein succinimidyl ester (CFSE) for the proliferation assays was purchased from Thermofisher (Darmstadt, Germany). The pokeweed (PWD) used was from Sigma-Aldrich (St Louis, MO, USA). To inhibit PD-L1/PD-1 crosstalk, the anti-PD-1 antibody used was from Bristol-Myers Squibb (Bristol, UK). The 2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamid (PCTA1) for the nuclear factor-κB (NF-κB) inhibitor assay was obtained from Tocris (Bristol, UK).