Statin-mediated Immunomodulation in Mice

Antibodies for in vivo mouse treatments specific for PD-1 (clone RMP1-14) and CD8 (clone YTS 169.4) were from BioXCell. Fluorescent-conjugated flow cytometry antibodies for mouse tumor experiments were obtained from BD Biosciences, Biolegend, Miltenyi or Abcam (see online supplemental methods table S1). Statin drugs (simvastatin, lovastatin, fluvastatin, pitavastatin, atorvastatin, rosuvastatin, pravastatin, table 1), were obtained from VWR International. For in vitro experiments, stock solutions were made with sterile DMSO and frozen at −20°C until use. For in vivo administration, statins were prepared as an oral suspension with 6% polyethylene glycol 400, 1% propylene glycol, and 0.1% Tween (all from Sigma) in water, as previously described.31 (link) Aliquots of statin suspension were frozen at −20°C for up to 2 weeks prior to administration to mice by oral gavage.