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Novi farmakološki pristopi za blokado nevrofilnih zunajceličnih pasti = Novel pharmacological approaches to block neutrophil extracellular trap formation : magistrski študijski program Laboratorijska biomedicina
Miholič, Marta
Neutrophils are phagocytic polymorphonuclear leukocytes that play an important role in the rapid innate host defence. They demonstrate their antimicrobial activity through phagocytosis, oxidative ...burst, and the formation of neutrophil extracellular traps. Neutrophil extracellular traps are extracellular structures consisting of DNA and granule proteins, which can capture and destroy invading pathogens. It was shown that neutrophils require glycolytic ATP, cytoskeleton rearrangement and ROS production for the generation of extracellular traps, but the mechanism by which the mitochondrial DNA gets released remains unexplained. SNARE proteins have previously been shown to play a part in neutrophil degranulation by mediating the transport and fusion of the granules to the plasma membrane. Here, we aim to investigate their role in mitochondrial DNA release and neutrophil extracellular trap formation. To evaluate their involvement in the neutrophil extracellular trap formation, we measured dsDNA release, degranulation, and ROS production in the presence of two fusion proteins, TAT-Syntaxin-4 and TAT-SNAP-23. Additionally, we performed the same assays in the presence of punicalagin, a known membrane stabilization agent, to investigate its effect on neutrophil extracellular trap formation. We confirm that Syntaxin-4 and SNAP-23 are necessary for the degranulation of primary and secondary granules of GM-CSF/C5a stimulated neutrophils. Furthermore, we demonstrate the involvement of SNARE proteins in neutrophil ROS production. Our findings, however, refute the notion that Syntaxin-4 and SNAP-23 are responsible for the release of mitochondrial DNA into extracellular space. Lastly, we demonstrate that punicalagin acts on the cytoskeleton, inhibiting dsDNA release and neutrophil extracellular trap formation.
Type of material - master's thesis
; adult, serious
Publication and manufacture - Ljubljana : [M. Miholič], 2022
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