Inflammation resolution circuits are uncoupled in acute sepsis and correlate with clinical severity
Bakr Jundi, Do-Hyun Lee, Hyungkook Jeon, Melody G. Duvall, Julie Nijmeh, Raja-Elie E. Abdulnour, Mayra Pinilla-Vera, Rebecca M. Baron, Jongyoon Han, Joel Voldman, Bruce D. Levy
Bakr Jundi, Do-Hyun Lee, Hyungkook Jeon, Melody G. Duvall, Julie Nijmeh, Raja-Elie E. Abdulnour, Mayra Pinilla-Vera, Rebecca M. Baron, Jongyoon Han, Joel Voldman, Bruce D. Levy
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Research Article Inflammation Pulmonology

Inflammation resolution circuits are uncoupled in acute sepsis and correlate with clinical severity

Abstract

Sepsis is a critical illness characterized by dysregulated inflammatory responses lacking counter-regulation. Specialized proresolving mediators are agonists for antiinflammation and for promoting resolution, and they are protective in preclinical sepsis models. Here, in human sepsis, we mapped resolution circuits for the specialized proresolving mediators resolvin D1 and resolvin D2 in peripheral blood neutrophils and monocytes, their regulation of leukocyte activation and function ex vivo, and their relationships to measures of clinical severity. Neutrophils and monocytes were isolated from healthy subjects and patients with sepsis by inertial microfluidics and resolvin D1 and resolvin D2 receptor expression determined by flow cytometry. The impact of these resolvins on leukocyte activation was determined by isodielectric separation and leukocyte function by stimulated phagolysosome formation. Leukocyte proresolving receptor expression was significantly higher in sepsis. In nanomolar concentrations, resolvin D1 and resolvin D2 partially reversed sepsis-induced changes in leukocyte activation and function. Principal component analyses of leukocyte resolvin receptor expression and responses differentiated sepsis from health and were associated with measures of sepsis severity. These findings indicate that resolvin D1 and resolvin D2 signaling for antiinflammation and resolution are uncoupled from leukocyte activation in early sepsis and suggest that indicators of diminished resolution signaling correlate with clinical disease severity.

Authors

Bakr Jundi, Do-Hyun Lee, Hyungkook Jeon, Melody G. Duvall, Julie Nijmeh, Raja-Elie E. Abdulnour, Mayra Pinilla-Vera, Rebecca M. Baron, Jongyoon Han, Joel Voldman, Bruce D. Levy

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Figure 2

RvD1 and RvD2 limit PMN activation by IDS in sepsis and healthy individuals.

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RvD1 and RvD2 limit PMN activation by IDS in sepsis and healthy individu...
(A) Experimental workflow for the isolation and assessment of PMN activation using isodielectric separation (IDS) to determine their electrical signature, isodielectric position (IDP). After isolation, PMN were first incubated with vehicle (<0.01% v/v EtOH), RvD1 (100 nM), or RvD2 (100 nM) for 15 minutes; they were then either stimulated by PMA (20 nM, red) or nonstimulated (black) for 30 minutes. Their IDPs were measured at 7 MHz frequency voltage. (B) Histogram plots of IDP distributions representative of healthy subjects (top panel) and patients with sepsis (bottom 3 panels). The dashed lines on the histogram plots are the median IDP, and their shift is determined upon stimulation of cells with PMA (red lines). (C) Box and whisker plots (median, 25th, and 75th percentiles) of the median IDP in nonstimulated (black) or PMA-stimulated (red) PMN of healthy subjects and patients with sepsis. (D) Box and whisker plots (median, 25th, and 75th percentiles) of the concentration-response curve of the median IDP of PMA-activated PMN from healthy subjects with exogenous RvD1 and RvD2. †P < 0.05 for nonstimulated veh versus stimulated veh in healthy subjects by paired, 2-tailed t test. *P < 0.05 for nonstimulated veh-health versus nonstimulated veh-sepsis by unpaired, 2-tailed t test. ††P < 0.05 for stimulated veh-health versus stimulated veh-sepsis by unpaired, 2-tailed t test. **P < 0.05 for unstimulated RvD1 versus nonstimulated veh in sepsis by unpaired, 2-tailed t test. †††P < 0.05 for stimulated RvD1 and RvD2 versus nonstimulated vehicle in health or sepsis by paired, 2-tailed t test. ***P < 0.05 for concentration-response curve of median IDP of stimulated PMN from healthy donors with exogenous RvD1, RvD2, and vehicle by 1-way ANOVA. n = 5 healthy subjects, n = 4 sepsis.