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 |
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Collection:
| Collection ID: | CO002218 |
| Collection Summary: | Both female and male mice were used in the experiments. Mice were maintained on a 12-h light/dark cycle with free access to food and water according to governmental rules. K320E-TWINKLE floxed mice (Baris et al., 2015) were crossed to CD23 CRE mice (Kwon et al., 2008) kindly provided by Meinrad Busslinger) to generate DNT animals. DNT mice used in these experiments had the genetic background DNT+/- CRE+/- and CRE control mice were DNT-/- CRE+/-. The WT animals used in this study were DNT-/- CRE-/- littermates. All mice were on the C57Bl/6 background. Isolation of primary murine cells from spleen and bone marrow Spleen was transferred into cold 2 % FCS (in PBS) and gently passed through a 70 µm cell strainer (BD) using the plunger of a 5 ml syringe (BD). Femur and tibia were flushed with cold 2 % FCS using a 27 G cannula (BD). Cell suspensions were pelleted by centrifugation at 300 x g for 5 min at 4°C. Erythrocytes were lysed in red blood cell-lysis buffer (150 mM NH4Cl, 10 mM KHCO3, 100 µM EDTA) for 5min at room temperature. The reaction was stopped by adding cold 2% FCS before centrifugation at 300 x g for 5 min at 4°C. The final cell suspensions were kept in cold 2 % FCS after filtration through 30 µm mesh filter (Sysmex). In vitro cultivation of primary murine B cells Splenic B cells were cultured with a starting concentration of 0.5 x 106 cells/ ml in R10 medium (RPMI1640, 10 % fetal calf serum (FCS), 2 mM glutamate, 1 mM sodium pyruvate, 50 U/ml penicillin G, 50 μg/ml streptomycin, 50 μM β-mercaptoethanol) for 72 h at 37°C and 5% CO2, supplemented with 10 µg/ml LPS. For in vitro class switch recombination cells were seeded at 0.1 x 106 cells/ ml in R10 medium for 96 h, supplemented with 5 ng/ml transforming growth factor , 5 nM retinoic acid, 10 µg/ml anti-CD40 antibody, 10 µg/ml LPS, 100 U/ml IL4 and 10 ng/ml IL5. Ref.: Baris, O.R., Ederer, S., Neuhaus, J.F., von Kleist-Retzow, J.C., Wunderlich, C.M., Pal, M., WunderlichF.T., Peeva, V., Zsurka, G., Kunz, W.S., et al. (2015). Mosaic Deficiency in Mitochondrial Oxidative Metabolism Promotes Cardiac Arrhythmia during Aging. Cell Metab 21, 667–677. |
| Collection Protocol Filename: | Bcellscoll Mielenz.pdf |
| Sample Type: | B-cells |
