We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
A novel approach to identifying and quantifying neutrophil extracellular trap formation in septic dogs using immunofluorescence microscopy.
- Authors
Li, Ronald H. L.; Johnson, Lynelle R.; Kohen, Casey; Tablin, Fern
- Abstract
Background: Canine neutrophils release neutrophil extracellular traps (NETs) in response to lipopolysaccharide but NETs from clinical septic dogs had not been identified. The primary aim is to describe the methodology of identifying and quantifying neutrophil extracellular traps (NETs) in cytology samples of septic foci in dogs with sepsis using immunofluorescence microscopy. Cytology samples including endotracheal tracheal wash (ETW), bronchoalveolar lavage (BAL), abdominal and pleural effusion collected from 5 dogs (3 septic, 2 non-septic) were fixed, permeabilized and stained for myeloperoxidase (MPO), citrullinated histone H3 (citH3) and cell-free DNA (cfDNA). Fluorescence microscopy was used to identify and quantify NETs in 10 random views at 40× magnification. NETs were identified based on co-localization of MPO, citH3 and cfDNA. NETs were quantified as a ratio (number of NETs: number of neutrophils). Neutrophils were identified based on cytoplasmic MPO, cellular diameter and nuclear morphology. Results: NETs were identified and quantified in all cytology samples collected from septic dogs. A small number of NETs was documented in one dog with sterile chronic bronchitis. No NETs were found in sterile abdominal effusion collected from one dog with congestive heart failure. Conclusions: Immunofluorescence microscopy could be a useful tool for the study of NETs in dogs with clinical sepsis.
- Subjects
NEUTROPHILS; SEPSIS; DOG diseases; MICROSCOPY; IMMUNOFLUORESCENCE
- Publication
BMC Veterinary Research, 2018, Vol 14, Issue 1, pN.PAG
- ISSN
1746-6148
- Publication type
Article
- DOI
10.1186/s12917-018-1523-z