Whole-body insulin sensitivity and metabolic responses to epinephrine are reduced during immunoactivation in several animal models, but effects of a chronic LPS infusion on these metabolic changes in lactating cows are not well-known. Therefore, experimental objectives were to characterize the metabolic response to an i.v. glucose tolerance test (GTT; 250 mg/kg BW) and epinephrine challenge (EC; 1.4 µg/kg BW) on d 8 of continuous LPS infusion in mid-lactation Holstein cows (164 ± 22 DIM; 676 ± 16 kg BW; parity 3.1 ± 0.4). Following 3 d acclimation, cows were enrolled in 2 experimental periods (P). During P1 (3 d), cows were fed ad libitum and baseline data were collected. Starting P2 (8 d), cows were assigned to 1 of 2 treatments: (1) saline-infused and pair-fed (CON-PF; i.v. 40 mL/h saline; n = 5) or (2) LPS-infused and ad libitum-fed (LPS-AL; E. coli O55:B5; 0.017, 0.020, 0.026, 0.036, 0.055, 0.088, 0.148, and 0.148 µg/kg BW/h for d 1–8, respectively; n = 6). LPS reduced DMI (28%) on d1 but DMI gradually returned to pre-infusion levels by the end of P2, and overall milk yield was reduced similarly (17%) between treatments. Following the d 8 0600h milking, feed was removed and all cows received a GTT. Feed was returned following the challenge. Four hours following the GTT feed was again removed and cows received an EC. Blood samples for both challenges were collected −30, −20, −10, 0, 5, 10, 20, 30, 45, 60, and 90 min relative to bolus. An additional 2.5 min postbolus sample was collected for the EC. For the GTT, area under the curve (AUC) was calculated through the 90 and 45 min sample for glucose and insulin, respectively. For the EC, AUC was calculated through the 60 min sample for glucose and nonesterified fatty acids. In response to the GTT, glucose and insulin AUC were increased 33 and 56%, respectively in LPS-AL relative to CON-PF cows (P = 0.01). There were no differences in the glucose or nonesterified fatty acid response to the EC. In summary, chronic LPS exposure induces peripheral insulin resistance but does not affect adipocyte and hepatocyte sensitivity to an epinephrine challenge.