Summary of Study ST003627

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002240. The data can be accessed directly via it's Project DOI: 10.21228/M8325D This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php

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Study IDST003627
Study TitleLipidomic analysis of human regulatory T cells.
Study SummaryRegulatory T cells (Tregs) are characterized by stable FOXP3 expression and controlling immune response by suppressive activity. Unique metabolic properties of Tregs are shown such as reduced glycolysis and increased oxidative phosphorylation. We have combined transcriptomics, metabolomics and lipidomics to dissect metabolic dynamics of Tregs upon activation. Combined metabolomic and lipidomic analysis showed that freshly isolated Tregs have unique metabolomic property, on the other hand, activated Tregs have unique lipidomic property. Interestingly, activated Tregs contained omega-3 polyunsaturated fatty acids (PUFA) enriched diglycerides and triglycerides.
Institute
Jikei University School of Medicine
Last NameSato
First NameYohei
Address3-25-8 Nishishinbashi, Minato-ku, Tokyo, 1058461, Japan
Emailyoheisatoo@gmail.com
Phone+81-3-3433-1111
Submit Date2024-12-09
Analysis Type DetailLC-MS
Release Date2025-06-02
Release Version1
Yohei Sato Yohei Sato
https://dx.doi.org/10.21228/M8325D
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR002240
Project DOI:doi: 10.21228/M8325D
Project Title:Lipidomic analysis of human regulatory T cells.
Project Summary:Regulatory T cells (Tregs) are characterized by stable FOXP3 expression and controlling immune response by suppressive activity. Unique metabolic properties of Tregs are shown such as reduced glycolysis and increased oxidative phosphorylation. We have combined transcriptomics, metabolomics and lipidomics to dissect metabolic dynamics of Tregs upon activation. Combined metabolomic and lipidomic analysis showed that freshly isolated Tregs have unique metabolomic property, on the other hand, activated Tregs have unique lipidomic property. Interestingly, activated Tregs contained omega-3 polyunsaturated fatty acids (PUFA) enriched diglycerides and triglycerides.
Institute:The Jikei University School of Medicine
Last Name:Sato
First Name:Yohei
Address:3-25-8 Nishishinbashi, Minato-ku, Tokyo, 1058461, Japan
Email:yoheisatoo@gmail.com
Phone:+81-3-3433-1111

Subject:

Subject ID:SU003757
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Group
SA393430Activated Teff1Activated Teff
SA393431Activated Teff2Activated Teff
SA393432Activated Teff3Activated Teff
SA393433Activated Treg1Activated Treg
SA393434Activated Treg2Activated Treg
SA393435Activated Treg3Activated Treg
SA393436Teff1Teff
SA393437Teff2Teff
SA393438Teff3Teff
SA393439Treg1Treg
SA393440Treg2Treg
SA393441Treg3Treg
Showing results 1 to 12 of 12

Collection:

Collection ID:CO003750
Collection Summary:Human regulatory T cells were sorted according to the CD4+CD25+CD127- populations by flow cytometry. Human effector T cells were sorted according to the CD4+CD25-CD127+ populations by flow cytometry. Cells were directly sorted in RPMI medium supplemented with 10% FBS. Spin down the collected cells and cell pellets were stored at -80°C.
Sample Type:T-cells

Treatment:

Treatment ID:TR003766
Treatment Summary:Human regulatory T cells were sorted and activated overnight by CD3/CD28/CD2 stimulation in the presence of 100U/mL recombinant human IL-2.

Sample Preparation:

Sampleprep ID:SP003764
Sampleprep Summary:Briefly, methanol was added to the frozen cell pellets to prepare a cell suspension. Total lipids were extracted from the entire suspension (equivalent to 1 x 10-6 cells) using the Bligh & Dyer method (partially modified), a liquid-liquid distribution method using chloroform, methanol, water, etc. The extracted lipid fraction was dried with nitrogen gas, then redissolved in 1 mL of methanol and transferred to a measurement vial for analysis.

Combined analysis:

Analysis ID AN005958 AN005959
Chromatography ID CH004528 CH004528
MS ID MS005673 MS005674
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Dionex Ultimate 3000 Thermo Dionex Ultimate 3000
Column L-column3 C18 Metal Free Column (100 x 2.0 mm, 2.0 μm) CERI L-column3 C18 Metal Free Column (100 x 2.0 mm, 2.0 μm) CERI
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Plus Orbitrap Thermo Q Exactive Plus Orbitrap
Ion Mode POSITIVE NEGATIVE
Units arbitrary units arbitrary units

Chromatography:

Chromatography ID:CH004528
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:L-column3 C18 Metal Free Column (100 x 2.0 mm, 2.0 μm) CERI
Column Temperature:40°C
Flow Gradient:Mobile phase A (isopropanol/methanol/water (5/1/4 v/v/v) supplemented with 5 mM ammonium formate and 0.05% ammonium hydroxide (28% in water))/mobile phase B (isopropanol supplemented with 5 mM ammonium formate and 0.05% ammonium hydroxide (28% in water)) ratios of 60%/40% (0 min), 40%/60% (0-1 min), 20%/80% (1-9 min), 5%/95% (9-11 min), 5%/95% (11-22 min), 95%/5% (22-22.1 min), 95%/5% (22.1-25 min), 60%/40% (25-25.1 min) and 60%/40% (25.1-30 min).
Flow Rate:0.1 mL/min
Solvent A:50% Isopropyl alcohol/10% Methanol/40% Water; 5mM Ammonium Formate; 0.05% Ammonium hydroxide
Solvent B:100% Isopropyl alcohol; 5mM Ammonium Formate, 0.05% Ammonium hydroxide
Chromatography Type:Reversed phase

MS:

MS ID:MS005673
Analysis ID:AN005958
Instrument Name:Thermo Q Exactive Plus Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:LC-ESI/MS/MS. The lipid identification analysis software Lipid Search 5. 1 (Mitsui Knowledge Industry Co., Ltd.) was used to identify lipid molecular species and align the measured samples. Correction value was calculated by following method (Total area correction). Correlation value = (Area value of detected peak) divided by (Total area value of detected peaks). A comprehensive lipidome analysis was performed on the received samples using IC-ESIMS/MS. The m/z values obtained and fragment ions were compared with the database registered in Lipid Search, and the number of molecular species that met the adoption criteria (our non-disclosed indicators) was 23 classes, 611 molecular species. The peak area of each identified molecule was corrected by the total area value (the sum of the detected area values). In addition, the relative values were used to correct the peak area.
Ion Mode:POSITIVE
  
MS ID:MS005674
Analysis ID:AN005959
Instrument Name:Thermo Q Exactive Plus Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:LC-ESI/MS/MS. The lipid identification analysis software Lipid Search 5. 1 (Mitsui Knowledge Industry Co., Ltd.) was used to identify lipid molecular species and align the measured samples. Correction value was calculated by following method (Total area correction). Correlation value = (Area value of detected peak) divided by (Total area value of detected peaks). A comprehensive lipidome analysis was performed on the received samples using IC-ESIMS/MS. The m/z values obtained and fragment ions were compared with the database registered in Lipid Search, and the number of molecular species that met the adoption criteria (our non-disclosed indicators) was 23 classes, 611 molecular species. The peak area of each identified molecule was corrected by the total area value (the sum of the detected area values). In addition, the relative values were used to correct the peak area.
Ion Mode:NEGATIVE
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