Journal Article

In vitro induction of NETosis: Comprehensive live imaging comparison and systematic review

van Wamel, Willem J. B., Hoppenbrouwers, Tamara, Abraham, Tsion E., van Cappellen, Wiggert A., Houtsmuller, Adriaan B., van Beusekom, Heleen M. M., de Maat, Moniek P. M., Autar, Anouchska S. A., van Neck, Johan W., Sultan, Andi R.

2017

Overview

Multiple inducers of in vitro Neutrophil Extracellular Trap (NET) formation (NETosis) have been described. Since there is much variation in study design and results, our aim was to create a systematic review of NETosis inducers and perform a standardized in vitro study of NETosis inducers important in (cardiac) wound healing. In vitro NETosis was studied by incubating neutrophils with PMA, living and dead bacteria (S. aureus and E. coli), LPS, (activated) platelets (supernatant), glucose and calcium ionophore Ionomycin using 3-hour periods of time-lapse confocal imaging. PMA is a consistent and potent inducer of NETosis. Ionomycin also consistently resulted in extrusion of DNA, albeit with a process that differs from the NETosis process induced by PMA. In our standardized experiments, living bacteria were also potent inducers of NETosis, but dead bacteria, LPS, (activated) platelets (supernatant) and glucose did not induce NETosis. Our systematic review confirms that there is much variation in study design and results of NETosis induction. Our experimental results confirm that under standardized conditions, PMA, living bacteria and Ionomycin all strongly induce NETosis, but real-time confocal imaging reveal different courses of events.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Antiinfectives and antibacterials / Aspergillosis / Bacteria / Bactericidal activity / Biocompatibility / Biology and Life Sciences / Blood / Calcimycin / Calcium / Calcium buffering / Calcium phosphates / Carbohydrates / Cardiology / Cardiovascular diseases / Centrifugation / Defense mechanisms / Defensive behavior / Deoxyribonucleic acid / Dilution / DNA / E coli / Erythrocytes / Escherichia coli - physiology / Ethylenediaminetetraacetic acids / Extracellular matrix / Extracellular Traps / Fluorescent Antibody Technique / Fungi / Health aspects / Heart diseases / Hematology / Humans / Hydrogen peroxide / In Vitro Techniques / In vivo methods and tests / Incubation / Infectious diseases / Inflammation / Interleukin 6 / Iodides / Literature reviews / Lungs / Markers / Medical research / Medicine and Health Sciences / Microbiology / Milk / Myocardium - pathology / Neonates / Neutrophils / Nuclei / Pathogens / Pathology / Phosphates / Physical Sciences / Physiological effects / Physiology / Protein kinase / Proteins / Research and Analysis Methods / Respiratory tract / Respiratory tract diseases / Staphylococcus aureus - physiology / Surgery / Temperature effects / Thrombosis / Toxicity / Wound Healing