In this thesis I study relation between hypoxia and histone H3
citrullination in neutrophilic granulocytes. Neutrophils are
innate immune cells and they often perform their effector
functions in hypoxic environment, such as a site of
inflammation. Histone citrullination is an important epigenetic
modification and it occurs at histone H3 in the greatest
extent. In the case of neutrophils, citrullination is mostly
associated with the process of NETosis, which is an important
neutrophil killing mechanism. For the study of the effect of
hypoxia, the isolated human neutrophils were incubated with
DMOG (dimethyloxalylglycine), which caused stabilisation of
HIF-α (hypoxia inducible factor). That switched neutrophils to
the hypoxic mode. Citrullinated histone H3 was determined by
western blot, intracellular ATP level was also monitored. The
results show that DMOG-induced hypoxia leads to histone H3
citrullination. Citrullination was observed already after 2
hours of incubation and the most intensive citrullination was
observed when 2 mM DMOG was used. In addition, I observed that
intracellular ATP concentration probably does not affect
histone H3 citrullination, but further research is needed to
completely elucidate the mechanism of action.