Influence of Different Bacteria Strains Isolated from Septic Children on Release and Degradation of Extracellular Traps by Neutrophils from Healthy Adults

Abstract

Neutrophils are the first line of immune defense against pathogens. They use three major antimicrobial mechanisms: phagocytosis, degranulation, and release of neutrophil extracellular traps (NETs). NETs are structures which consist of nuclear DNA conjugated with antibacterial proteins. They are formed to entrap and kill pathogens. The aim of the study was to evaluate the influence of Escherichia coli (E. coli), Streptococcus pneumoniae (S. pneumoniae), Stenotrophomonas maltophilia (S. maltophilia), and Pseudomonas aeruginosa (P. aeruginosa), isolated from the peripheral blood of children with sepsis, on the release and degradation of NETs by neutrophils isolated from blood healthy adult subjects. Neutrophils were stimulated with the bacterial strains outlined above. The quantitative and qualitative analyses of NETs release were performed by fluorometric measurement and immunofluorescence, respectively. The ability of bacteria to degrade NETs was studied qualitatively. Oxidative burst was assessed by flow cytometry. Histone H3 citrullination was evaluated by Western blot. We found that NETs were formed only when neutrophils were incubated with S. pneumoniae. E. coli, P. aeruginosa, and S. maltophilia did not induce the release of the NETs. P. aeruginosa, S. pneumoniae, and E. coli induced the production of reactive oxygen species (ROS) by neutrophils. Two studied bacterial strains (S. pneumoniae and E. coli) were able to degrade NETs. However, none of the strains induced the citrullination of histone H3. We conclude that the ability of bacteria to induce and degrade NETs depends on the specific bacterial strain.