Summary of Study ST002140
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 PR001355. The data can be accessed directly via it's Project DOI: 10.21228/M8K70K 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.
| Study ID | ST002140 |
| Study Title | Mitochondrial respiration in B lymphocytes is essential for humoral immunity by controlling flux of the TCA cycle |
| Study Summary | The function of mitochondrial respiration during B cell fate decisions and differentiation 55 remained equivocal. This study reveals that selection for mitochondrial fitness occurs during B 56 cell activation and is essential for subsequent plasma cell differentiation. By expressing a 57 mutated mitochondrial helicase in transitional B cells, we depleted mitochondrial DNA during 58 B cell maturation, resulting in reduced oxidative phosphorylation. Although no changes in 59 follicular B cell development were evident, germinal centers, class switch recombination to 60 IgG, plasma cell maturation and humoral immunity were diminished. Defective oxidative 61 phosphorylation led to aberrant flux of the tricarboxylic acid cycle and lowered the amount of 62 saturated phosphatidic acid. Consequently, mTOR activity and BLIMP1 induction were 63 curtailed whereas HIF1 _and glycolysis were amplified. Exogenous phosphatidic acid 64 increased mTOR activity in activated B cells. Hence, mitochondrial function is required and 65 selected for in activated B cells for the successful generation of functional plasma cells. |
| Institute | University of Erlangen-Nürnberg |
| Department | Division of Molecular Immunology.Universitätsklinikum Erlangen, Nikolaus Fibinger Zentrum |
| Laboratory | Prof. Mielenz |
| Last Name | Mielenz |
| First Name | Dirk |
| Address | Nikolaus-Fiebiger-Zentrum, Glückstraße 6, 91054 Erlangen |
| dirk.mielenz@fau.de | |
| Phone | ++49 9131 8539105 |
| Submit Date | 2022-04-06 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS/MS(Dir. Inf.) |
| Release Date | 2022-05-02 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN003501 | AN003502 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Ion Chromatography | None (Direct infusion) |
| Chromatography system | ThermoDionexICS3000 | TriVersa NanoMate |
| Column | ThermoDionexAS11/AG11 | TriVersa NanoMate |
| MS Type | ESI | ESI |
| MS instrument type | QTRAP | QTRAP |
| MS instrument name | ABI Sciex 3200 QTrap | ABI Sciex 6500 QTrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | mol% | pmol/10E6 cells |
Chromatography:
| Chromatography ID: | CH002587 |
| Chromatography Summary: | The HPLC-system was controlled by the software Chromeleon VS 6.8 and DCMS-Link VS1.1 (Dionex) in combination with Analyst 1.4.1 (Applied Biosystems). Metabolites were separated on two IonPac AS11HC columns (2 × 250 mm; Dionex) protected by an AG11HC guard column (2 × 50 mm). The elution gradient was generated with water (eluent A) and 100 mm KOH (eluent B) within a total run time of 80 min at a flow rate of 0.25 mL min−1 and a column temperature of 35°C as follows: 0 min, 4%; 0 to 1 min, 4%; 1 to 6 min, 15%; 6 to 12 min, 19%; 12 to 22 min, 20%; 22 to 24 min, 23%; 24 to 27 min, 35%; 27 to 37 min, 38%; 37 to 39 min, 45%; 39 to 44 min, 100%; 44 to 71 min, 100%; 71 to 76 min, 4%; and 76 to 80 min, 4% eluent B. Ref.: Hofmann, J., Bornke, F., Schmiedl, A., Kleine, T., and Sonnewald, U. (2011). Detecting functional groups of Arabidopsis mutants by metabolic profiling and evaluation of pleiotropic responses. 10Front Plant Sci 2, 82. |
| Instrument Name: | ThermoDionexICS3000 |
| Column Name: | ThermoDionexAS11/AG11 |
| Chromatography Type: | Ion Chromatography |
| Chromatography ID: | CH002588 |
| Chromatography Summary: | Dried lipid extracts were dissolved in 300 μl of methanol. 20 μl of the lipid extract in methanol were loaded into 96-well plates and diluted with 20 μl of 20 mM ammonium acetate in methanol. Lipid infusion and ionization was conducted using Nano-ESI chips with the TriVersa NanoMate operated by the ChipSoft Software (Advion) under the following settings: sample infusion volume: 14 μl, volume of air to aspirate after sample: 1 μl, air gap before chip: enabled, aspiration delay: 0 s, prepiercing: with mandrel, spray sensing: enabled, cooling temperature: 14°C, gas pressure: 0.5 psi, ionization voltage: 1.4 kV, and vent headspace: enabled. Prewetting was done once. Ref.: Kumar, V., Bouameur, J.E., Bar, J., Rice, R.H., Hornig-Do, H.T., Roop, D.R., Schwarz, N., Brodesser, 1120 S., Thiering, S., Leube, R.E., et al. (2015). A keratin scaffold regulates epidermal barrier 1121 formation, mitochondrial lipid composition, and activity. J Cell Biol 211, 1057–1075. Ref.: Kumar, V., Bouameur, J.E., Bar, J., Rice, R.H., Hornig-Do, H.T., Roop, D.R., Schwarz, N., Brodesser, 1120 S., Thiering, S., Leube, R.E., et al. (2015). A keratin scaffold regulates epidermal barrier 1121 formation, mitochondrial lipid composition, and activity. J Cell Biol 211, 1057–1075. |
| Instrument Name: | TriVersa NanoMate |
| Column Name: | TriVersa NanoMate |
| Chromatography Type: | None (Direct infusion) |
