Response to correspondence: Flow cytometric quantification of neutrophil extracellular traps: Limitations of the methodological approach by C iepiela et al.
Year: 2016
Type: article
Abstract: the average amount of neutrophils which expressed citH3 after 2 min was 7.54 6 4.46% and did not differ between unstimulated, PMA-or CI-treated cells (n56).There were no changes in the level of citH3 in unstimulated samples over time.The amount of citH3 increased in samples stimulated with PMA (7.25 6 4.8%, 8.55 6 5.0%, 10.2 6 5.8% in 2nd, 5th, and 10th min, respectively) with significant difference between 2nd and 10th min (p50.03).In CI-stimulated samples, the level of citH3 also increased to 7.7 6 5.0, 9.03 6 5.8, 9.03 6 3.8% over time, but there was no statistical significance (p50.06 and 0.09 for 2nd vs 5th and 2nd vs 10th min) (Fig. 1B).The level of citrullination after 10th min did not change significantly up to the 3rd h of incubation with NETs' stimulators.The flow cytometry method for NETs' quantification is a real challenge as it allows to analyze cells only in suspension.Undoubtedly, Gavillet et al. opened a new field in NETs' studies.However, the methodology of flow-assisted extracellular traps' analysis needs significant improvement.Recently, Zhao et al. published a paper based on imaging flow cytometry, which allows for parallel quantification and morphological assessment of NETs.Their analysis is based on the assessment of decondensed nuclei in the cells as a first step in NET formation and analysis of elongation of cells' shape [5].Nowadays, it seems to be the most suitable NETs' quantification method for rapid studies in a suspension of cells.
Source: American Journal of Hematology
DOI: 10.1002/ajh.24292
Authors: David A. Williams, Mathilde Gavillet
Language: English
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W2225577511