Peculiarities of Neutrophil Extracellular Traps Formation in Chinchilla Rabbits

Abstract

Neutrophil extracellular traps (NETs) are decondensed nuclear chromatin decorated with bactericidal proteins of various cell organelles and performing an effector function aimed at combating pathogens in the inflammatory focus. At the same time, NETs play an important role in the pathogenesis of many autoimmune and inflammatory diseases as well as malignancies. Rabbits are one of the most commonly used species of laboratory animals in medical and biological research. A large number of models of various diseases of the cardiovascular, immune and other human systems have been developed on rabbits. However, in the scientific literature, there is no information about the ability of rabbit neutrophils to undergo NETosis (the programmed cell death in the process of NETs formation) in response to well-known pharmacological stimuli. The purpose of the present study was to investigate in the in vitro system the ability of neutrophils of Soviet chinchilla rabbit to form NETs in response to mimetic of diacylglycerol phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. To isolate rabbit neutrophils, the Ficoll-Hypaque one-step density gradient centrifugation with modifications was used. Oxidative burst was assessed with luminol-amplified chemiluminescence method, while NETs formation was assessed with immunofluorescence analysis. The study shows for the first time that neutrophils of Soviet chinchilla rabbit do not form NETs in response to PMA but do form traps in response to A23187, and have low levels of oxidative burst in response to PMA, A23187 and chemoattractant N-formyl-methionyl-leucyl-phenylalanine.