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- Title
Characterization of Early-Phase Neutrophil Extracellular Traps in Urinary Tract Infections.
- Authors
Yu, Yanbao; Kwon, Keehwan; Tsitrin, Tamara; Bekele, Shiferaw; Sikorski, Patricia; Nelson, Karen E.; Pieper, Rembert
- Abstract
Neutrophils have an important role in the antimicrobial defense and resolution of urinary tract infections (UTIs). Our research suggests that a mechanism known as neutrophil extracellular trap (NET) formation is a defense strategy to combat pathogens that have invaded the urinary tract. A set of human urine specimens with very high neutrophil counts had microscopic evidence of cellular aggregation and lysis. Deoxyribonuclease I (DNase) treatment resulted in disaggregation of such structures, release of DNA fragments and a proteome enriched in histones and azurophilic granule effectors whose quantitative composition was similar to that of previously described in vitro-formed NETs. The effector proteins were further enriched in DNA-protein complexes isolated in native PAGE gels. Immunofluorescence microscopy revealed a flattened morphology of neutrophils associated with decondensed chromatin, remnants of granules in the cell periphery, and myeloperoxidase co-localized with extracellular DNA, features consistent with early-phase NETs. Nuclear staining revealed that a considerable fraction of bacterial cells in these structures were dead. The proteomes of two pathogens, Staphylococcus aureus and Escherichia coli, were indicative of adaptive responses to early-phase NETs, specifically the release of virulence factors and arrest of ribosomal protein synthesis. Finally, we discovered patterns of proteolysis consistent with widespread cleavage of proteins by neutrophil elastase, proteinase 3 and cathepsin G and evidence of citrullination in many nuclear proteins.
- Subjects
NEUTROPHILS; URINARY tract infections; ANTI-infective agents; DEOXYRIBONUCLEASES; FLUORESCENCE microscopy; DNA; STAPHYLOCOCCUS aureus genetics; BACTERIAL genetics; ESCHERICHIA coli; GENETICS
- Publication
PLoS Pathogens, 2017, Vol 13, Issue 1, p1
- ISSN
1553-7366
- Publication type
Article
- DOI
10.1371/journal.ppat.1006151