Transition cows experience a nutrient deficit, particularly immediately postpartum. The inflammatory balance is altered at this time, and cows exhibit an immune response primed for inflammation. Studies in other mammals demonstrate a role of nutrient-sensing kinases in determining immune phenotypes and inflammatory balance. Our primary objective was to investigate changes in energy storage (glycogen) and signaling through the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) and adenosine monophosphate-activated kinase (AMPK) pathway in bovine immune cells in the transition period. Glycogen concentration was measured by an enzymatic-fluorometric method in peripheral blood mononuclear cells (PBMC) of multiparous Holstein cows (n = 72) at 3 time points (21 d before, 7 and 28 d after calving). Phosphorylation of AKT, mTOR substrates 4EBP1 and S6RP, and AMPK were assessed by immunoblotting in PBMC from 60 animals at the same time. Repeated measures ANOVA was performed in Proc MIXED (SAS v. 9.4) and pairwise comparisons controlled with Tukey’s procedure. The average (95% CI) concentration of glycogen decreased from 1.5 (1.35 to 1.66) µg/10⁶ PBMC on d 21 prepartum to 63% of the prepartum levels on d 7 postpartum [0.95 (0.86 to 1.04) µg/10⁶ PBMC, P < 0.0001)]. On d 28 postpartum, average concentrations increased to 84% of the prepartum values [1.25 (1.12 to 1.41) µg/10⁶ PBMC], P < 0.01 to both other time points. Compared with prepartum values, activation of AKT in bovine PBMC on d 7 was reduced by 9% (P = 0.001), non-phosphorylated 4EBP1 increased by 5% (P = 0.07) whereas no change was observed in the phosphorylation of S6RP (P > 0.27). The activation of AMPK increased by 42% postpartum (P < 0.001). We conclude that decreasing energy storage at the time of the nadir in nutrient balance goes along with changes in nutrient-signaling pathways in bovine PBMC postpartum. Temporal associations with changes in immune response at this time and the documented role of the studied pathways in immune-phenotype determination of other species are intriguing and warrant further investigation as a possible link between nutrient deficit, metabolism, and bovine immune response postpartum.