Pantoprazole

The trial was approved by the medical agencies, the ethical committees and the data agency, and was conducted in accordance with the Helsinki declaration and good clinical practice as described by Danish law (ClinicalTrials.gov ID: NCT00937625).
Patients were admitted to hospital at day −8 and a central venous catheter was applied. A lymphodepleting chemotherapy regimen consisting of Cyclophosphamide, 60 mg/kg/d day −7 to −6 and fludarabine phosphate, 25 mg/m2/d at day −5 to −1 administered as previously described [10 (link)]. Prophylactic antiemetics with palonosetron, aprepitant, and Domperidone were given together with pantoprazole. At day 0 autologous TILs were infused intravenously followed by 14 days of subcutaneous IL-2 injections, 2 MIU, starting the same evening.
Patients were treated prophylactically with trimethoprim, sulfamethoxazole, and acyclovir from the beginning of treatment and 6 months thereafter and with fluconazole during the leucopenic period.
Clinical response was monitored with a computed tomography (CT) or positron emission tomography (PET)/CT scan 8 weeks after T cell infusion and assessed according to RECIST 1.0. All scans were reviewed by an independent radiologist at the Hospital who was blinded to previous descriptions.

HEK cells (400000 cells/well) were seeded into 24-well cell culture plates (Greiner Bio-One, Frickenhausen, Germany) and grown for 48 h. Cell adherence was improved by coating plates with poly-L-lysine (Biochrom AG, Berlin, Germany) prior to cell plating. All uptake studies were carried out at 37°C as described [31] (link). For uptake measurements, cells were firstly washed with uptake buffer (145 mM NaCl, 5 mM hydroxyethylpiperazine ethanesulfonic acid, 3 mM KCl, 1 mM CaCl₂, 0.5 mM MgCl₂, 5 mM glucose, pH 7.4, [46] (link)) prewarmed to 37°C. Uptake was initiated by replacing this solution with uptake buffer containing 5 µM [¹⁴C]metformin, 100 µM [¹⁴C]TEA, 10 µM [¹⁴C]omeprazole, or 10 µM [¹⁴C]pantoprazole. Uptake was stopped at indicated time points by removing the uptake buffer and immediately washing the cells 3 times with ice-cold uptake buffer. Cells were lysed with 0.2% SDS and intracellular radioactivity was determined by liquid scintillation counting (Hidex 300SL TDCR liquid scintillation counter, Turku, Finland). For measuring inhibition of OCT-mediated metformin uptake by PPIs, 5 µM [¹⁴C]metformin uptake was measured in the presence of different PPI concentrations and stopped after 5 min (OCT1, OCT3) or 5 sec (OCT2).
For determination of intracellular accumulation of unlabeled lansoprazole, rabeprazole, and tenatoprazole, cells were incubated for 1 or 5 min with 5 µM PPI at 37°C and then immediately washed three times with ice-cold uptake buffer and twice with ice-cold phosphate-buffered saline. Cells were harvested by scraping them off in 500 µl 0.1 M sodium carbonate buffer (pH 9.3) : methanol 4∶1 (v/v) and then transferred into Eppendorf tubes. Cells were disrupted by three cycles of shock freezing/thawing (liquid nitrogen, 37°C water bath) and further by ultra-sonification, three times 3 sec, at 4°C. Disrupted cell solutions were centrifuged 5 min (15000 g) at 4°C and supernatants were transferred into Eppendorf tubes and stored at −20°C for analytic determination of the PPIs.
Protein content of lysed cells was determined in 25-µl aliquots using the bicinchoninic acid assay as described [31] (link).