It is known that exercise through endogenous catecholamines such as adrenaline (Adr) and noradrenaline (Nor) can have a significant effect on number and activity of immune cells, including the neutrophils. The mechanisms of these influences have not been studied sufficiently. The aim of the present study was to investigate in vitro effect of Adr, Nor and selective agonist of β-adrenergic receptor isoproterenol (Iso) on neutrophil functional activity and cytokine production in athletes. Ethical approval for the study of neutrophils from 29 healthy athletes of both sexes (19.5±1.14 years old) was granted by the University Ethics Committee. The functional activity of neutrophils was determined in EDTA-stabilized venous blood. We defined the capacity of neutrophils to ingest zymosan in phagocytosis assay, oxidative burst in spontaneous and stimulated NBT-test. Before the tests, the blood was incubated (37°C, 1 h) with 100 μl of DPBS or Adr (0.01, 0.1, 1 μM), Nor (0.001, 0.1, 1 μM), Iso (0.1 μM). To analyse how catecholamines effect on cytokine secreting activity, neutrophils were isolated by two different methods: (1) by density gradient centrifugation of whole blood over medium Lympholyte-H, dextran sedimentation and red cells osmotic lysis (Neu1); (2) similarly to Neu1, but with an additional immunomagnetic negative selection step, using the EasySep Human Neutrophil Enrichment Kit (StemCell Technologies) (Neu2). Purity of the isolated neutrophils was assessed by Romanowsky-Giemsa staining and flow cytometry using markers CD66b-FITC/CD16-PE. The purity of neutrophil suspension averaged 92-98% for Neu1 and >99% for Neu2. Neutrophils were resuspended in RPMI 1640 medium and incubated (37°C, 5% CO2, 20 h) at a concentration of 1×106 cells/ml/well with addition of 100 μl DPBS or 100 μl of different agonists of adrenoreceptors. Concentrations IL-1b, IL-6, IL-8, TNF-a and MIP-1b in cell-free supernatants were measured by ELISA kits. Statistical analysis was carried out using non-parametric models. We found that Adr and Nor in the pharmacological concentration (1 μM) inhibited neutrophil phagocytic and fMLP-induced oxidative activity (P<0.05). Adr in all doses stimulated the secretion of IL-1b by Neu1 (P<0.05). Stimulation of IL-1b secretion was also observed after the incubation of Neu1 with Iso (P<0.05). Nor did not effect on neutrophil cytokine release. A basal secretion level of IL-1b and MIP-1b was not detected in supernatants from non-stimulated neutrophils after additional immunomagnetic negative selection (Neu2). Unlike Neu1 Adr and Iso did not stimulate release IL-1b by Neu2. Taken together, our results indicate that Adr and Nor in the pharmacological concentration have anti-inflammatory effect on neutrophil phagocytic and oxidative activity, but they have limited effects on the secretion of cytokines by neutrophils in athletes.