Summary of Study ST001997

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench,, where it has been assigned Project ID PR001268. The data can be accessed directly via it's Project DOI: 10.21228/M8T39S This work is supported by NIH grant, U2C- DK119886.


This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001997
Study TitlePolyamine import and accumulation causes immunomodulation in macrophages engulfing apoptotic cells (Part 2)
Study SummaryPhagocytosis of apoptotic cells, termed efferocytosis, is critical for tissue homeostasis and drives anti-inflammatory programming in engulfing macrophages. Here, we assess metabolites in naïve and inflammatory macrophages following engulfment of multiple cellular and non-cellular targets. Efferocytosis leads to unique increases in the arginine-derived polyamines, spermidine and spermine, in vitro and in vivo. Surprisingly, polyamine accumulation after efferocytosis does not arise from retention of apoptotic cell metabolites or de novo synthesis, but from enhanced polyamine import that is dependent on Rac1, actin, and PI3 kinase. Blocking polyamine import prevents efferocytosis from suppressing macrophage IL-1or IL-6. This identifies efferocytosis as a trigger for polyamine import and accumulation, and imported polyamines as mediators of efferocytosis-induced immune reprogramming.
University of Colorado Denver
Last NameHaines
First NameJulie
Address12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA
Submit Date2021-11-22
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2021-12-08
Release Version1
Julie Haines Julie Haines application/zip

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Collection ID:CO002071
Collection Summary:Experimental animals This study was approved and performed in accordance with the ethical guidelines of the Institutional Animal Care and Use Committee at National Jewish Health. C57BL/6J (000664), B6-CD45.1 (002014) and CCR2KO (004999) mice were obtained from Jackson Laboratories (Bar Harbor, ME, USA). DsRed.T3 mice (available as 006051 at Jackson Labs) were generously shared by the Nagy laboratory (Vintersten et al., 2004). CCR2KO mice were crossed with DsRed mice until double homozygous. Mice used in experiments were between 8-16 weeks of age. Both male and female sex were used in experiments except bone marrow chimera experiments, which used only male mice. Human tissue This study used deidentified donor peripheral blood mononuclear cells (PBMC) collected and isolated (Young et al., 2011) by the National Jewish Blood Prep Core (IRB No. HS-1285). Age and sex of deidentified blood donors in this study is not known. Cell Lines Jurkat (human T lymphocytes, clone E6-1) were obtained from ATCC and cultured in RPMI-1640 (GIBCO) supplemented with 10% heat-inactivated fetal bovine serum (v/v) (GeminiBio), 1X Pen/Strep/Glut, 1mM HEPES, and 100M Sodium Pyruvate. Cells were cultured in a humidified CO2 incubator at 37C. The Jurkat cell line was established from peripheral blood of a 14-year-old male diagnosed with T cell leukemia. Murine primary cell cultures Peritoneal cells were isolated from naïve mice by lavage with 10mL of PBS containing 0.5mM EDTA. Lavage was estimated to contain 50% macrophages. Peritoneal macrophages were isolated by adhesion purification of lavage cells in RPMI-10 (RPMI-1640 containing 10% heat-inactivated fetal bovine serum (v/v), 1X Pen/Strep/Glut, 1mM HEPES, and 100M Sodium Pyruvate) plated 1.3x106 macrophages/well in 6-well plates for FACS sorting or 4x106 macrophage/well in 24-well plates for flow cytometry. Non-adherent cells were removed by washing after 3-5 hours, and macrophages were cultured in fresh RPMI-10 overnight. 400ng/mL purified E. coli O111:B4 LPS (List Biological Laboratories Inc.) was added to generate inflammatory macrophages 12h before the addition of target cells. Bone marrow-derived macrophages were generated by 8-day culture of murine bone marrow cells grown in media containing M-CSF (High glucose DMEM containing 10% v/v FBS, 20% v/v L929-conditioned media, 1X Pen/Strep/Glut, 100M Sodium Pyruvate, 55M beta-Mercaptoethanol). All macrophages were removed from culture dishes by 3-minute incubation with 1X Trypsin-EDTA (Sigma) plus gentle scraping.
Sample Type:Macrophages