Summary of Study ST003626
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 PR002239. The data can be accessed directly via it's Project DOI: 10.21228/M86R70 This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST003626 |
| Study Title | Metabolomic profile of human regulatory T cells. |
| Study 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 | Jikei University School of Medicine |
| Last Name | Sato |
| First Name | Yohei |
| Address | 3-25-8 Nishishinbashi |
| yoheisatoo@gmail.com | |
| Phone | +81-3-3433-1111 |
| Submit Date | 2024-12-09 |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-06-02 |
| Release Version | 1 |
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Project:
| Project ID: | PR002239 |
| Project DOI: | doi: 10.21228/M86R70 |
| Project Title: | Metabolomic profile 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 |
| Email: | yoheisatoo@gmail.com |
| Phone: | +81-3-3433-1111 |
Subject:
| Subject ID: | SU003756 |
| 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 |
|---|---|---|
| SA393418 | Activated Teff1 | Activated T-cells(Teff) |
| SA393419 | Activated Teff2 | Activated T-cells(Teff) |
| SA393420 | Activated Teff3 | Activated T-cells(Teff) |
| SA393421 | Activated Treg1 | Activated T-cells(Treg) |
| SA393422 | Activated Treg2 | Activated T-cells(Treg) |
| SA393423 | Activated Treg3 | Activated T-cells(Treg) |
| SA393424 | Teff1 | T-cells(Teff) |
| SA393425 | Teff2 | T-cells(Teff) |
| SA393426 | Teff3 | T-cells(Teff) |
| SA393427 | Treg1 | T-cells(Treg) |
| SA393428 | Treg2 | T-cells(Treg) |
| SA393429 | Treg3 | T-cells(Treg) |
| Showing results 1 to 12 of 12 |
Collection:
| Collection ID: | CO003749 |
| 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: | TR003765 |
| 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: | SP003763 |
| Sampleprep Summary: | After culturing the regulatory T cells, the medium was removed and washed twice with mannitol (WAKO pure chemical). A methanol (WAKO pure chemical) solution was added and stirred. Milli-Q water containing 10 μM of an internal standard (HMT, Patent No. 6173667) was added and stirred, and centrifuged (2,300 × g, 4°C, 5 min) (Patent No. 6173667). After centrifugation, the extract was transferred to an ultrafiltration tube (Ultrafree MC PLHCC, HMT, centrifugal filter unit 5 kDa). This was centrifuged (9,100 × g, 4°C) and subjected to ultrafiltration. The filtrate was dried and dissolved in Milli-Q water again for measurement. |
Combined analysis:
| Analysis ID | AN005956 | AN005957 |
|---|---|---|
| Chromatography ID | CH004526 | CH004527 |
| MS ID | MS005671 | MS005672 |
| Analysis type | MS | MS |
| Chromatography type | CE | CE |
| Chromatography system | Agilent 7100 CE | Agilent 7100 CE |
| Column | Fused silica capillary i.d. 50 µm x 80cm | Fused silica capillary i.d. 50 µm x 80cm |
| MS Type | ESI | ESI |
| MS instrument type | TOF | TOF |
| MS instrument name | Agilent 6230 TOF | Agilent 6230 TOF |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | arbitrary number | arbitrary units |
Chromatography:
| Chromatography ID: | CH004526 |
| Instrument Name: | Agilent 7100 CE |
| Column Name: | Fused silica capillary i.d. 50 µm x 80cm |
| Column Temperature: | N/A |
| Flow Gradient: | N/A |
| Flow Rate: | N/A |
| Solvent A: | Cation Buffer Solution (p/n:H3301-1001) |
| Solvent B: | N/A |
| Chromatography Type: | CE |
| Chromatography ID: | CH004527 |
| Instrument Name: | Agilent 7100 CE |
| Column Name: | Fused silica capillary i.d. 50 µm x 80cm |
| Column Temperature: | N/A |
| Flow Gradient: | N/A |
| Flow Rate: | N/A |
| Solvent A: | Anion Buffer Solution(p/n:I3302-1023) |
| Solvent B: | N/A |
| Chromatography Type: | CE |
MS:
| MS ID: | MS005671 |
| Analysis ID: | AN005956 |
| Instrument Name: | Agilent 6230 TOF |
| Instrument Type: | TOF |
| MS Type: | ESI |
| MS Comments: | CE-TOFMS(Basic) positive(Cation) mode. The detected peaks were matched and searched against all substances registered in the HMT metabolite library based on the m/z and MT values. The search tolerance was set to ± 0.5 min for MT and ± 10 ppm for m/z. Mass error (ppm) = (Actual value – theoretical value)/ Actual value×106 For substances with molecular weights of 100 or less and some substances expected to be present in large amounts in the sample, the tolerance was set wider than the above conditions and the search was performed. In addition, if the candidates could not be narrowed down and the same candidate metabolite was assigned to multiple peaks, a branch number was added and the list was displayed. The target metabolites were analyzed. The calibration curve was made using peak areas corrected by the internal standard substance, and the concentration was calculated for each substance with a single calibration of 100 μM (internal standard substance 200 μM). |
| Ion Mode: | POSITIVE |
| MS ID: | MS005672 |
| Analysis ID: | AN005957 |
| Instrument Name: | Agilent 6230 TOF |
| Instrument Type: | TOF |
| MS Type: | ESI |
| MS Comments: | CE-TOFMS(Basic) negative(Anion) mode. The detected peaks were matched and searched against all substances registered in the HMT metabolite library based on the m/z and MT values. The search tolerance was set to ± 0.5 min for MT and ± 10 ppm for m/z. Mass error (ppm) = (Actual value – theoretical value)/ Actual value×106 For substances with molecular weights of 100 or less and some substances expected to be present in large amounts in the sample, the tolerance was set wider than the above conditions and the search was performed. In addition, if the candidates could not be narrowed down and the same candidate metabolite was assigned to multiple peaks, a branch number was added and the list was displayed. The target metabolites were analyzed. The calibration curve was made using peak areas corrected by the internal standard substance, and the concentration was calculated for each substance with a single calibration of 100 μM (internal standard substance 200 μM). |
| Ion Mode: | NEGATIVE |
