Changes in Neutrophil Metabolism upon Activation and Aging

Neutrophil activation is an important mechanism of host defense against pathogens. Chronic inflammation and autoimmunity are often associated with abnormalities in phenotype and functions of neutrophils. Since effector functions of immune cells during inflammation are tightly linked to their metabolic state, changes in neutrophil metabolome upon activation have been investigated in this study. Human neutrophils from healthy blood donors (n = 6) were treated either with tumor necrosis factor α (TNF-α) or lipopolysaccharide (LPS), whereas untreated neutrophils were used as control. Since apoptotic cells are abundant at sites of inflammation, the metabolome of aged, mainly apoptotic neutrophils was analyzed too. NMR spectroscopy of water-soluble metabolites revealed a clear distinction between aged neutrophils and neutrophils in control and activated samples. Higher levels of NAD⁺ (4- to 9-fold) and lower levels of ATP (0.3-fold), glutathione (0.8-fold), hypotaurine (0.8-fold), and phosphocholine (0.6-fold) were detected in aged neutrophils than in the other samples. Differences in metabolic profiles between LPS and TNF-α-stimulated cells as well as between stimulated and control neutrophils were statistically not significant. Replication with additional six blood donors confirmed increased NAD⁺ levels in aged cells compared to activated and control neutrophils.