Assessment of metabolic activity and energy supply of peripheral blood lymphocytes

Abstract

BACKGROUND: T-cells have the capability to change their metabolism in response to activation signals. Resting T-cells primarily use the oxidation of higher fatty acids and oxidative phosphorylation in mitochondria for their energy needs, whereas activated T-cells switch to aerobic glycolysis and glutaminolysis, using glucose, and glutamine as substrates, respectively.
 AIM: To determine the metabolic activity and energy supply of peripheral blood lymphocytes in predominantly healthy northerners by measuring the intracellular content of HIF-1 (hypoxia-induced factor 1), SIRT3 (sirtuin 3), and ATP (adenosine triphosphate).
 MATERIALS AND METHODS: 39 volunteers, residents of the Arkhangelsk region (23 women and 16 men, 2362 years old), were selected, and examined for this experiment. We established the total number of peripheral blood lymphocytes with CD-typing of lymphocytes (CD3+, CD4+, CD8+, CD71+) by indirect immunoperoxidase reactions, the content of HIF-1 and SIRT3 in the lymphocyte lysate employing enzyme immunoassay, the concentration of ATP by luminescent analysis through luciferin-luciferase reaction. Statistical analysis was conducted in "Statistica 10.0", cluster analysis was applied using the k-means method. Mean values (M) and standard deviations (SD) were calculated, the normal distribution was tested by the KolmogorovSmirnov and Lilliefors criterion. Student's t-test was calculated, and the differences were considered statistically significant at p 0.05.
 RESULTS: The study revealed that the metabolic activity of lymphocytes associated with HIF-1 regulation differs significantly in the examined volunteers,, while in the group with a lower total number of lymphocytes and their subpopulations (CD3+, CD4+, CD8+, CD71+) there is a predominant glycolytic orientation of metabolism with proliferated cells energy supply.
 CONCLUSION: Metabolic activity which can be determined by the HIF-1/SIRT3 ratio, and the energy supply of lymphocytes have a substantial impact on their differentiation, proliferation, and functioning.