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Objective. The objective of our research is to study the rate and modes of cell death among white blood cells using an Annexin V and 7-AAD flow cytometry kit in patients with chronic rhinosinusitis without nasal polyps (CRSsNP). Methods. Flow cytometry was used to detect cell death modes of leukocytes in twelve patients with CRSsNP. Annexin V FITC, anti-CD45 PE and 7-AAD (7-aminoactinomycin D) were used for the assessment of apoptosis stages. The current method allows identifying four different states of cells: 1 viable cells (Annexin Vnegative,7-AAD negative cells); 2 early apoptotic cells (AnnexinV positive, 7-AAD negative cells); 3 late apoptotic/necrotic cells (AnnexinV positive, 7AAD positive cells); 4 dead necrotic cells (AnnexinV negative, 7-AAD positive cells). To assess the rate of oxidative stress in CRSsNP, 8-isoprostane levels in blood serum were determined using ELISA. Results. We found that CRSsNP was accompanied by the development of oxidative stress, evidenced by elevation of 8isoprostane in blood serum, and the increased rate of cell death among white blood cells with the prevalence of early apoptotic cells, confirmed by a higher percentage of AnnexinV positive, 7-AAD negative cells compared with control subjects. Conclusion. Our findings suggest that CRSsNP is accompanied by the activation of leukocyte apoptosis.

Distinguishing modes of leukocyte cell death using flow cytometry in patients with chronic rhinosinusitis without nasal polyps