Resumen
Neutrophil extracellular traps (NETs) are extracellular web-like structures comprised of proteins and DNA that have been associated with the development of cancer. Citrullination of histone H3 has been implicated in the formation of NETs. The aim of our study was to develop a quick and reliable method for NET quantitation. Here, we describe a novel protocol for the quantification of NETs based on the detection of citrullinated histone H3 bound to DNA (CitH3DNA binding assay). This assay was validated by comparing the ability of neutrophils from control tumor-free and myeloma-bearing mice to form NETs in response to stimuli. We demonstrated that neutrophils from tumor-bearing mice produced more NETs than those from tumor-free counterparts following stimulation with PMA. The increase in NET production as detected by microscopy correlated with significantly higher histone H3 citrullination levels and increased measurements of CitH3DNA in our novel binding assay.