The shifting lipidomic landscape of blood monocytes and neutrophils during pneumonia
Alex R. Schuurman, … , W. Joost Wiersinga, Tom van der Poll
Alex R. Schuurman, … , W. Joost Wiersinga, Tom van der Poll
Published February 22, 2024
Citation Information: JCI Insight. 2024;9(4):e164400. https://doi.org/10.1172/jci.insight.164400.
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Research Article Immunology Infectious disease

The shifting lipidomic landscape of blood monocytes and neutrophils during pneumonia

Abstract

The lipidome of immune cells during infection has remained unexplored, although evidence of the importance of lipids in the context of immunity is mounting. In this study, we performed untargeted lipidomic analysis of blood monocytes and neutrophils from patients hospitalized for pneumonia and age- and sex-matched noninfectious control volunteers. We annotated 521 and 706 lipids in monocytes and neutrophils, respectively, which were normalized to an extensive set of internal standards per lipid class. The cellular lipidomes were profoundly altered in patients, with both common and distinct changes between the cell types. Changes involved every level of the cellular lipidome: differential lipid species, class-wide shifts, and altered saturation patterns. Overall, differential lipids were mainly less abundant in monocytes and more abundant in neutrophils from patients. One month after hospital admission, lipidomic changes were fully resolved in monocytes and partially in neutrophils. Integration of lipidomic and concurrently collected transcriptomic data highlighted altered sphingolipid metabolism in both cell types. Inhibition of ceramide and sphingosine-1-phosphate synthesis in healthy monocytes and neutrophils resulted in blunted cytokine responses upon stimulation with lipopolysaccharide. These data reveal major lipidomic remodeling in immune cells during infection, and link the cellular lipidome to immune functionality.

Authors

Alex R. Schuurman, Osoul Chouchane, Joe M. Butler, Hessel Peters-Sengers, Sebastiaan Joosten, Xanthe Brands, Bastiaan W. Haak, Natasja A. Otto, Fabrice Uhel, Augustijn Klarenbeek, Christine C.A. van Linge, Antoine van Kampen, Mia Pras-Raves, Michel van Weeghel, Marco van Eijk, Maria J. Ferraz, Daniël R. Faber, Alex de Vos, Brendon P. Scicluna, Frédéric M. Vaz, W. Joost Wiersinga, Tom van der Poll

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Figure 6

Inhibition of SPT and Sphk1 blunts LPS-induced cytokine release by monocytes and neutrophils ex vivo.

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Inhibition of SPT and Sphk1 blunts LPS-induced cytokine release by monoc...
(A) Box-and-whisker plots depicting cytokine release of monocytes (500,000 cells per well, 10 healthy donors) preincubated for 1 hour with either myriocin or PF543, and subsequently stimulated for 24 hours with lipopolysaccharide (LPS; 100 ng/mL). The y axis depicts concentration in pg/mL (±SEM, vertical bars). The colors indicate the different conditions. Significance was determined by a paired Wilcoxon’s rank-sum test. A P value below 0.05 was considered significant. *P < 0.05, **P < 0.01. (B) Identical to panel A, but here for neutrophils (500,000 cells per well, 8 healthy donors), stimulated for 2 hours with LPS. (C) Box-and-whisker plots showing levels (in pmol/mL) of ceramide species in monocytes incubated with either vehicle or myriocin, measured by targeted MS (n = 4 per condition). Cer, ceramide; dh-Cer, dihydroceramide; glcCer, glucosylceramide; lacCer, lactosylceramide; Gb3, globotriaosylceramide. (D) Box-and-whisker plots showing levels (in pmol/mL) of S1P in monocytes and neutrophils incubated with either vehicle or PF543, measured by targeted MS (n = 4 per condition). S1P, sphingosine-1-phosphate. (E) Box-and-whisker plots showing intensity levels of intracellular phosphorylation markers p-ERK-171Yb (pERK), p-NF-κB-Er166 (p-NF-κB), and p-p38-Gd156 (pp38) in monocytes and neutrophils after LPS stimulation and treatment with either myriocin or PF543, measured by cytometry by time of flight (CyTOF). The y axis indicates the arsinh-normalized medium intensity levels. There were no significant differences (n =4 per condition). (F) Box-and-whisker plots showing mRNA levels for key cytokines in monocytes treated with vehicle, myriocin, or PF543, measured by qRT-PCR. There were no significant differences (n = 4 per condition).