DETERMINATION OF ACTIVATION OF PHAGOCYTES IN MOUSE BLOOD

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Authors

KOCURKOVÁ Anna KUBALA Lukáš

Year of publication 2020
Type Chapter of a book
MU Faculty or unit

Faculty of Science

Citation
Description Phagocytes play a key role in inflammation, a complex process underlying the pathogenesis of numerous chronic and acute diseases. Determination of phagocyte activation provides an essential information about inflammatory processes. Among strategies of how to estimate phagocyte activation status is determination of surface expression of selected receptors and adhesion molecules reflecting degranulation of phagocytes. Herein, the activation of mouse blood neutrophil granulocytes was analyzed in mice treated by schizophyllan (SPG), a (1-3)­-beta-D-glucan produced as a cell wall constituent of fungi or plants with suggested immunomodulatory potential. The activation of neutrophil granulocytes was analyzed 2, 4, 6 and 8 hours after SPG i.v. application. Staining of cells with antibodies against Ly6G, CDllb and CD62l was performed in whole blood with consequent fixation and lysation of erythrocytes. Neutrophil granulocytes were identified based on typical forward and side scatter characteristics and expression of Ly6G surface antigen. Further, pro-inflammatory cytokine, TNF-alpha, was analyzed in plasma by ELISA. Application of SPG significantly increased expression of CDllb after 4 hours. CD62l was significantly decreased at 4 and 6 hours after SPG application. Overall, it can be concluded that SPG induced activation of neutrophil granulocytes in mice after intravenous application in time frame from 2 to 8 hours with the highest peak after 4 hours and that CDllb and CD621 are sensitive markers that can be applied for determination of activation of this abundant phagocyte population.
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