A cluster of immunoresolvents links coagulation to innate host defense in human blood

Abstract

Blood coagulation is a protective response that prevents excessive bleeding upon blood vessel injury. Here, we investigated the relationship between coagulation and the resolution of inflammation and infection by lipid mediators (LMs) through metabololipidomic-based profiling of human whole blood (WB) during coagulation. We identified temporal clusters of endogenously produced pro-thrombotic and proinflammatory LMs (eicosanoids), as well as specialized proresolving mediators (SPMs). In addition to eicosanoids, a specific SPM cluster was identified that consisted of resolvin E1 (RvE1), RvD1, RvD5, lipoxin B4, and maresin 1, each of which was present at bioactive concentrations (0.1 to 1 nM). Removal of adenosine from the coagulating blood markedly enhanced the amounts of SPMs produced and further increased the biosynthesis of RvD3, RvD4, and RvD6. The cyclooxygenase inhibitors celecoxib and indomethacin, which block the production of thomboxanes and prostanoids, did not block the production of clot-driven SPMs. Unbiased mass cytometry analysis demonstrated that the SPM cluster produced in human blood targeted leukocytes at the single-cell level, directly activating ERK and CREB signaling in neutrophils and CD14+ monocytes. Treatment of human whole blood with the components of this SPM cluster enhanced both the phagocytosis and killing of Escherichia coli by leukocytes. Together, these data identify a pro-resolving LM circuit, including endogenous molecular brakes and accelerators, which promoted host defense. These temporal LM-SPM clusters can provide accessible metabolomic profiles for precision and personalized medicine.

Maresin 1 activates LGR6 receptor promoting phagocyte immunoresolvent functions

“…CyTOF was also carried out with peripheral blood leukocytes (compare with ref. 39). MaR1 increased phosphorylation of pERK and pCREB, giving the strongest signals in PMN and CD14 + classical monocytes followed by CD16 + non-classical monocytes and dendritic cells.…”

Resolvins in inflammation: emergence of the pro-resolving superfamily of mediators

“…As with ALX/FPR2, peptide and SPM signaling events by this receptor have distinct patterns of activation for intracellular pathways, including ERK and NF-κB phosphorylation (46). SPM interactions with ALX/FPR2 and ERV1 decrease NF-κB activity and cytokine production (46,116). Translocation of NF-κB to the nucleus and its activity are also regulated by SPM signaling at DRV1 and DRV2 (97,100,117).…”

“…In human peripheral blood, several laboratories collectively identified plasma SPMs (38,168), as well as a plasma SPM cluster consisting of RvE1, RvE2, RvD1, 17-epi-RvD1, RvD2, RvD5, RvD6, PD1, 17-HDHA, and 18-HEPE (150,153), and a serum cluster of RvD1, 17-epi-RvD1, RvD2, RvD3, PD1, MaR1, RvE1, and RvE2. SPM concentrations attained in human peripheral blood target PMNs and monocytes (at the single-cell level determined by CyTOF mass cytometry) to increase phagocytosis and killing of E. coli (116). PUFAs are associated with reduced incidence of fatal coronary heart disease (169,170), and it has recently been established that omega-3 supplementation at doses up to 10 g/d (EPA and DHA) does not increase risk of bleeding or affect other coagulation parameters (75).…”

Astrocyte-derived lipoxins A4 and B4 promote neuroprotection from acute and chronic injury

“…Analysis was carried out in negative ion mode, and eicosanoids and PUFA were quantitated using scheduled multiple reaction monitoring (MRM) using from 4 to 6 specific transition ions for each analyte. We used specific retention time and established prominent fragment ions for identification and quantification of LXB 4 and LXA 4 (52,71,72 qPCR. Mouse retinal mRNA was isolated using RNeasy Isolation Kit (QIAGEN), quantified using NanoDrop, and mRNA reverse transcribed with the High Capacity cDNA Kit (Applied Biosystems).…”