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Adenosine from a biologic source regulates neutrophil extracellular traps (NETs).

Journal of leukocyte biology (2019-03-26)
Kai Xu, Kimberly A Cooney, Eric Y Shin, Lanfang Wang, Juline N Deppen, Sydney C Ginn, Rebecca D Levit

Neutrophil extracellular traps (NETs) are implicated in autoimmune, thrombotic, malignant, and inflammatory diseases; however, little is known of their endogenous regulation under basal conditions. Inflammatory effects of neutrophils are modulated by extracellular purines such as adenosine (ADO) that is inhibitory or ATP that generally up-regulates effector functions. In order to evaluate the effects of ADO on NETs, human neutrophils were isolated from peripheral venous blood from healthy donors and stimulated to make NETs. Treatment with ADO inhibited NET production as quantified by 2 methods: SYTOX green fluorescence and human neutrophil elastase (HNE)-DNA ELISA assay. Specific ADO receptor agonist and antagonist were tested for their effects on NET production. The ADO 2A receptor (A2A R) agonist CSG21680 inhibited NETs to a similar degree as ADO, whereas the A2A R antagonist ZM241385 prevented ADO's NET-inhibitory effects. Additionally, CD73 is a membrane bound ectonucleotidase expressed on mesenchymal stromal cells (MSCs) that allows manipulation of extracellular purines in tissues such as bone marrow. The effects of MSCs on NET formation were evaluated in coculture. MSCs reduced NET formation in a CD73-dependent manner. These results imply that extracellular purine balance may locally regulate NETosis and may be actively modulated by stromal cells to maintain tissue homeostasis.

Product Number
Product Description

Phorbol 12-myristate 13-acetate, ≥99% (TLC), film or powder
Lipopolysaccharides from Escherichia coli O111:B4, γ-irradiated, BioXtra, suitable for cell culture
Red Blood Cell Lysis Buffer, solution, Roche, pkg of 100 mL, sufficient for 50-500 reactions
Adenosine, ≥99%
Cell Death Detection ELISA, sufficient for ≤96 tests, suitable for ELISA, kit of 1 (9 components)
Ethylene glycol-bis(2-aminoethylether)-N,N,N′,N′-tetraacetic acid, BioXtra, ≥97 .0%
5′-(N-Ethylcarboxamido)adenosine, powder