Resolvin D2 is a potent regulator of leukocytes and controls microbial sepsis

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Abstract

A growing body of evidence indicates that resolution of acute inflammation is an active process1,2. Resolvins are a new family of lipid mediators enzymatically generated within resolution networks that possess unique and specific functions to orchestrate catabasis, the phase in which disease declines2,3. Resolvin D2 (RvD2) was originally identified in resolving exudates, yet its individual contribution in resolution remained to be elucidated. Here, we establish RvD2’s potent stereoselective actions in reducing excessive neutrophil trafficking to inflammatory loci. RvD2 decreased leukocyte–endothelial interactions in vivo by endothelial-dependent nitric oxide production, and by direct modulation of leukocyte adhesion receptor expression. In mice with microbial sepsis initiated by caecal ligation and puncture, RvD2 sharply decreased both local and systemic bacterial burden, excessive cytokine production and neutrophil recruitment, while increasing peritoneal mononuclear cells and macrophage phagocytosis. These multi-level pro-resolving actions of RvD2 translate to increased survival from sepsis induced by caecal ligation and puncture and surgery. Together, these results identify RvD2 as a potent endogenous regulator of excessive inflammatory responses that acts via multiple cellular targets to stimulate resolution and preserve immune vigilance.

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Figure 1: Stereochemical assignment, biosynthesis and total organic synthesis of RvD2.
Figure 2: RvD2 potently reduces leukocyte–endothelial interactions to reduce microbial peritonitis.
Figure 3: Modulation of leukocyte trafficking by RvD2 is nitric oxide dependent.
Figure 4: RvD2 reduces bacterial levels, systemic inflammation and enhances survival in microbial sepsis.

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Acknowledgements

We acknowledge support from National Institutes of Health grants GM-38765 and P50-DE016191 (C.N.S.), Welcome Trust Programme grant 086867/Z/08/Z (R.J.F. and M.P.) and Project grant 085903/Z/08 (R.J.F.) and Arthritis Research Campaign UK fellowships 18445 and 18103 (to L.V.N. and D.C., respectively). M.S. received a National Research Service Award from the NHLBI (HL087526). We thank J. W. Winkler and J. Uddin for work related to RvD2 synthesis, P. Pillai, K. Martinod, G. Fredman and J. Dalli for technical assistance, and M. H. Small for assistance with the manuscript. We also thank B. Schmidt for histopathology, Children’s Hospital Boston.

Author Contributions M.S. and L.V.N. designed and carried out experiments, analysed data and wrote the manuscript; L.S., R.Y. and D.C. carried out experiments and analysed data; N.A.P. synthesized RvD2; R.J.F. and M.P. designed experiments, analysed data and contributed to the manuscript; C.N.S. planned the project, designed experiments, analysed data and wrote the manuscript.

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Correspondence to Charles N. Serhan.

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Competing interests

[Competing Interests: The resolvins are biotemplates for stable analogues. Patents on these are awarded and assigned to Brigham and Women’s Hospital, C.N.S. is inventor. These analogue patents are licensed for clinical development.]

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-11 with Legends, Supplementary Methods and Supplementary References. (PDF 4216 kb)

Supplementary Movie 1

This file shows a time-lapse movie (approximately 1 min at double-speed) of leukocyte adhesion to a post-capillary venule, and emigration into the surrounding tissue following 1h of PAF (1 nM) superfusion. (MOV 3218 kb)

Supplementary Movie 2

This file shows a time-lapse movie (approximately 1 min at double-speed) of decreased leukocyte adhesion to a post-capillary venule, and emigration into the surrounding tissue following 2h of PAF (1 nM) plus 1h of RvD2 (1 nM) superfusion. (MOV 3289 kb)

Supplementary Movie 3

This file shows the remarkable difference in behaviour of vehicle (red) or RvD2 (100 ng i.v.; blue) treated mice 12h after CLP-induced sepsis. (MOV 5050 kb)

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