Induction of early-phase endotoxin tolerance in horses

Six, clinically healthy horses, of mixed age and sex, were infused via a jugular venous catheter with 100 ml of pyrogenfree sterile saline (PFSS; 0.9% NaCl). Animals were infused with Escherichia coli O55:B5 endotoxin (total dose = 50 ng/kg bwt), 24 (LPS-1) and 48 h (LPS-2) after PFSS infusion. Blood was collected before, and every 15 min after, each infusion for the first 8 h and then every 2 h for the following 14 h. Clinical responses (rectal temperature, heart rate, respiration rate and blood pressure) were determined before and every 4 h after each infusion for 20 h. Geometric mean anti-endotoxin antibody titres in serum samples, harvested just before each infusion, were unchanged over the course of the experiment. Serum tumour necrosis factor-alpha (TNF alpha) activity was estimated using a cytotoxic bioassay and WEHI 164 clone 13 murine fibrosarcoma cells as targets. Mean clinical parameter values and geometric mean serum TNF alpha activity at given time points were compared across the 3 infusions. Both LPS-1 and LPS-2 resulted in elevated mean rectal temperature at 4 h after infusion. However, duration of mean rectal temperature elevation was greater (P < 0.05) after LPS-1 (through 12 h) than after LPS-2 (through 8 h). More substantial increases in systolic and diastolic blood pressure were observed after LPS-1 than LPS-2 and mean systolic blood pressure after LPS-1 was elevated at 4 h when compared to PFSS (P < 0.05). Decreased systolic and diastolic blood pressures were observed at 16 h after both LPS infusions, when compared to PFSS infusion. Heart rate was increased, compared to PFSS, after both LPS-1 (8-12 h) and LPS-2 (4-12 h) (P < 0.05). No significant elevations in mean respiratory rate were observed after either LPS-1 or LPS-2 when compared to PFSS. However, at 4 h post infusion, mean respiratory rate after LPS-2 was greater (P < 0.05) than that after LPS-1. Serum TNF alpha activity was not detected after infusion of PFSS, but was detected after both LPS-1 and LPS-2. Serum TNF alpha activity was elevated earlier, was present in higher concentrations and persisted longer after LPS-1 than after LPS-2 (P < 0.05). The decreased duration of fever and attenuated serum TNF alpha response subsequent to successive sublethal LPS challenge observed in this study support the conclusion that these horses developed early-phase endotoxin tolerance (EPET) and, therefore, contributes to the understanding of the role of endotoxaemia in a number of clinical conditions in horses.