PAF-Induced Lethal Shock Model

A schematic schedule of the experimental procedure is shown in Figure ​Figure2A.2A. To determine the effect of PAF on the survival of ICR mice, we divided the animals (6 weeks of age) into the following three groups: PAF group, CpG-A₁₅₈₅ + PAF group, and CpG-A₁₅₈₅ + MAFP + PAF group (N = 8 mice per group). ICR mice were i.p. injected with 50, 100, or 300 µg CpG-A₁₅₈₅ 1, 3, and 5 days before PAF challenge. MAFP (1 mg/kg) was i.p. injected 20 min before CpG-A₁₅₈₅ injection. A stock solution of PAF was made in methanol, and the required aliquot was dried under a stream of nitrogen. PAF was then reconstituted in 0.5 ml PBS containing 0.1% human serum albumin (Wako Pure Chemical Industries, Ltd., Osaka, Japan) before use and administered i.p. into Swiss albino mice. After each treatment, animals were monitored for up to 60 min for survival.