Abt 737

Uninfected C57BL/6 mice received 5000 CD8⁺ T cells, isolated from CD45.1⁺ P14-Bim-mCherry mice with CD8a⁺ T-Cell Isolation Kit (Miltenyi Biotec Inc.), through intravenous (i.v.) adoptive transfer and were infected with LCMV one day later. On day 10 post-infection, splenic CD8⁺ T cells were enriched with CD8a⁺ T-Cell Isolation Kit. Bim-mCherry cells, P14-Bim-mCherry cells, or Nur77-GFP cells were further sorted on BD FACSAria II (BD Biosciences) with the assistance of the Research Flow Cytometry Core at CCHMC. One million of sorted CD8⁺ T cells were i.v. adoptive transferred into timed-infected congenic recipients. After 14 days, the lymphocytes in spleen were analyzed by flow cytometry. ABT-737 was a generous gift from Abbott Laboratories [65 (link)] and given to mice as previously described [15 (link), 22 (link), 66 (link)], 1 mg/mouse/day between days 14 and 23 post-infection.

ABT-737: The mice were treated with either 100 mg/kg/d ABT-737 solution or an equal volume of carrier solution administrated via intraperitoneal injections (i.p.) for 21 consecutive days. ABT-737 was received from Abbott (now AbbVie Inc). After the treatment the mice were followed up for 21 days. Navitoclax, venetoclax, metformin: The mice were treated with vehicle (5% EtOH, 20% Phosal PG in MQ), 100 mg/kg/d Navitoclax or venetoclax, 300 or 600 g/kg/d metformin or with drug combinations delivered via intragastric (i.g.) route for 21 days. The cohorts were followed up until 60 days after transplantation. Navitoclax and venetoclax were from AbbVie Inc. Drugs were sonicated daily for better solubility. In the experiments including anti-PD-1, mice received i.p. injections of either 200 µg control IgG (bxcell, #BE0089) or 200 µg anti-PD-1 (bxcell, #BE0146) every third day, in total four times, together with the 1-week daily adjuvant i.g. treatments of either vehicle, venetoclax, metformin or the combination of venetoclax+metformin. Paclitaxel was administrated every third day, i.p., 10 mg/kg in cremphor EL-ethanol-saline. Plasma ALAT levels were measured in the Biochemical Analysis Core for Experimental Research, University of Helsinki.