Date of Graduation

5-2025

Document Type

Thesis

Degree Name

Master of Science in Cell & Molecular Biology (MS)

Degree Level

Graduate

Department

Cell & Molecular Biology

Advisor/Mentor

Durdik, Jeannine M.

Committee Member

Ivey, D. Mack

Second Committee Member

Lessner, Daniel J.

Third Committee Member

Pare, Adam

Keywords

macrophage metabolism; fever response; glycolysis; oxidative phosphorylation; RNA sequencing; species-specific immune response

Abstract

We investigated the real-time metabolic rates of activated macrophage cell lines derived from mouse, chicken and rainbow trout. Each species showed a different kinetic signature with metabolic rates accelerating under fever conditions specific to each organism. Upon activation mouse macrophages increased their rate of glycolysis early and then a later reprogramming phase occurred with a dramatic decrease in oxidative phosphorylation (OXPHOS) accompanied by a reciprocal second increase in glycolytic rates. Chicken macrophages showed an initial increase in both glycolytic and OXPHOS rates. The late drop OXPHOS rates also occurred in chicken however there was no reprogramming that occurred as glycolytic rates steadily decreased. Although overall metabolic rates differed depending on stimuli and temperature, rainbow trout macrophages showed a response to temperature increases but weaker metabolic responses to activation. To determine the role of fever on the macrophage transcriptome, RNA sequencing was performed using mouse macrophages isolated from four time points, referred to as BANG, determined by distinct features in their metabolic signature. The results indicated that differential expression was most prominent at 39°C early after stimulation. It was confirmed that overall protein synthesis is upregulated when cells were exposed to fever temperatures. Additionally, fever showed increased regulation of inflammatory responses to lipopolysaccharide (LPS) either alone or with additional cytokine stimulus. The enrichment of the metabolic response showed both glycolysis and OXPHOS were differentially expressed early and at higher temperatures. This suggests that the acceleration of the metabolic response due to increased temperature is initiated early upon activation.

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