Summary of Study ST001653

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 PR001059. The data can be accessed directly via it's Project DOI: 10.21228/M8T69T This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

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 IDST001653
Study TitleMucosal-Associated-Invariant-T (MAIT) cells stimulated with IL-12/IL-12, anti-CD3/CD28 or both for 16 hours
Study SummaryThe untargeted metabolomics analysis was performed after metabolite extraction from vital cells. The main object of the study was to define the activation of MAIT cells on the molecular level.
Institute
Helmholtz Centre for Environmental Research
Last NameEngelmann
First NameBeatrice
AddressPermoserstraße 15, Leipzig, Saxony, 03418, Germany
Emailbeatrice.engelmann@ufz.de
Phone00493412351099
Submit Date2021-01-12
Num Groups4
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2021-02-01
Release Version1
Beatrice Engelmann Beatrice Engelmann
https://dx.doi.org/10.21228/M8T69T
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001059
Project DOI:doi: 10.21228/M8T69T
Project Title:Untargeted metabolomics analysis of Mucosal-Associated-Invariant-T (MAIT) cells stimulated with IL-12/IL-12, anti-CD3/CD28 or both
Project Type:Untargeted metabolomics
Project Summary:MAIT cells are unique for their ability to recognize bacterial metabolites, inducing an antigen(ag)-dependent activation, but can also be activated in an ag-independent manner but the molecular details of MAIT cell activation are not completely understood. In order to define the activation of MAIT cells on the molecular level, among other things we applied untargeted metabolomics.
Institute:Helmholtz Centre for Environmental Research - UFZ
Department:Molecular Systems Biology
Last Name:Engelmann
First Name:Beatrice
Address:Permoserstraße 15, Leipzig, Saxony, 03418, Germany
Email:beatrice.engelmann@ufz.de
Phone:00493412351099

Subject:

Subject ID:SU001730
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Cell Strain Details:CD161+ TCR V 7.2+ MAIT cells were obtained by positive magnetic separation after isolation of Peripheral blood mononuclear cells from male human donors.

Factors:

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

mb_sample_id local_sample_id Treatment
SA151753Spender60_CD3_pa-CD3/a-CD28 activation
SA151754Spender59_CD3_pa-CD3/a-CD28 activation
SA151755Spender53_CD3_pa-CD3/a-CD28 activation
SA151756Spender42_CD3_pa-CD3/a-CD28 activation
SA151757Spender49_CD3_pa-CD3/a-CD28 activation
SA151758Spender42_combi_pcombinated activation
SA151759Spender60_combi_pcombinated activation
SA151760Spender53_combi_pcombinated activation
SA151761Spender59_combi_pcombinated activation
SA151762Spender49_combi_pcombinated activation
SA151748Spender49_IL12_pIL12/IL18 activation
SA151749Spender60_IL12_pIL12/IL18 activation
SA151750Spender53_IL12_pIL12/IL18 activation
SA151751Spender59_IL12_pIL12/IL18 activation
SA151752Spender42_IL12_pIL12/IL18 activation
SA151763Spender59_unstim_punstimulated
SA151764Spender42_unstim_punstimulated
SA151765Spender49_unstim_punstimulated
SA151766Spender60_unstim_punstimulated
SA151767Spender53_unstim_punstimulated
Showing results 1 to 20 of 20

Collection:

Collection ID:CO001723
Collection Summary:Peripheral blood mononuclear cells were isolated from male human donors at age 20-50 by Ficoll-Paque™ density-gradient centrifugation. CD161+ TCR Valpha 7.2+ MAIT cells were obtained by positive magnetic separation after isolation of Peripheral blood mononuclear cells from male human donors.
Sample Type:Blood (whole)

Treatment:

Treatment ID:TR001743
Treatment Summary:MAIT cells were stimulated with 50ng/ml IL-12 and 50ng/ml IL-18, 10µg/ml plate-bound anti-CD3 and 1µg/ml soluble anti-CD28 or a combination of both.

Sample Preparation:

Sampleprep ID:SP001736
Sampleprep Summary:Cells were quenched 3 times with 1 ml ice-cold 0.9% sodium chloride. After removing the quenching solution, cells were resuspended in 100 µl ice-cold acetonitrile followed by 100 µl ice-cold Milli-Q water. After vortexing for 1 min, cells were centrifuged (14000 rpm, 4 °C, 10 min). Supernatants were transferred to new tubes. Intracellular metabolites were extracted again with 500 µl Methanol:ACN:Milli-Q water (2:3:1). After centrifugation (14000 rpm, 4 ° C, 10 min) both supernatants were combined, evaporated to dryness and stored at -80 °C until measurement.

Combined analysis:

Analysis ID AN002700
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1290
Column Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um)
MS Type ESI
MS instrument type QTOF
MS instrument name Agilent 6540 QTOF
Ion Mode POSITIVE
Units peak area

Chromatography:

Chromatography ID:CH001992
Instrument Name:Agilent 1290
Column Name:Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um)
Column Temperature:45
Flow Rate:0.3 ml/min
Sample Injection:10 µl
Solvent A:100% water; 0.1% formic acid
Solvent B:50% acetonitrile/50% MeOH; 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS002498
Analysis ID:AN002700
Instrument Name:Agilent 6540 QTOF
Instrument Type:QTOF
MS Type:ESI
MS Comments:Mass Hunter software was used to obtain raw data.Eluted metabolites were measured with the QTOF operated in centroid mode. Full scan data was generated with a scan range of 60-1600 m/z in positive ionization mode. Out of the survey scan the 5 most abundant precursor ions with charge state = 1 were subjected to fragmentation. The dynamic exclusion time was set at 30 s. Peak picking and database search was done using Progenesis QI software.
Ion Mode:POSITIVE
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