Summary of Study ST000657
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 PR000462. The data can be accessed directly via it's Project DOI: 10.21228/M8BG7G 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 | ST000657 |
| Study Title | mTOR regulates metabolic adaptation of APCs in the lung microenvironment and controls the outcome of allergic inflammation |
| Study Summary | Antigen presenting cells (APCs) occupy diverse anatomical tissues, but their tissue-restricted homeostasis remains poorly understood. Here, working in mouse models of inflammation, we found that mTOR-dependent metabolic adaptation was required at discrete locations. mTOR was dispensable for DC homeostasis in secondary lymphoid tissues, but necessary to regulate cellular metabolism and accumulation of CD103+ DCs and alveolar macrophages in lung. Moreover, whilst numbers of mTOR-deficient lung CD11b+ DCs were not changed, they were metabolically reprogrammed to skew allergic inflammation from eosinophilic Th2 to neutrophilic Th17 polarity. The mechanism for this change was independent of translational control, but dependent on inflammatory DC which produced IL-23 and increased fatty acid oxidation. mTOR therefore mediates metabolic adaptation of APCs in distinct tissues, influencing the immunological character of allergic inflammation. |
| Institute | Emory University |
| Last Name | Li; Gardinassi |
| First Name | Shuzhao; Luiz |
| Address | Emory Woodruff Memorial Research Building, 1639 Pierce Dr NE, Atlanta, GA 30322 |
| shuzhao.li@gmail.com; luiz.gardinassi@emory.edu | |
| Phone | 404-712-2988 or 404-727-5091 |
| Submit Date | 2017-06-21 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2018-07-17 |
| Release Version | 1 |
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Collection:
| Collection ID: | CO000674 |
| Collection Summary: | Lungs and Spleen of WT or mTOR-APC-KO mice were aseptically removed. Spleens were finely chopped with a scalpel in 6-well plates, and digested with 1-1.5mg/mL collagenase type 4 (Worthington Biochemical Corporation) dissolved in HBSS. Digestion was performed for 30 minutes at 37°C, and reactions were stopped with addition of 2mM EDTA. Digestions were passed through 40μm cell strainers to obtain a homogenous cell suspension. To obtain lung suspensions, mice were sacrificed with CO2 and perfused with 5mL PBS injected into the right ventricle. Lungs were dissected into gentleMACS™ C tubes (Miltenyi) and dissociated using a gentleMACS™ octo dissociator using the pre-set program m_lung_01_02. Dissociated lung was digested with 1-1.5mg/mL collagenase type 4 for 30 minutes at 37°C. Lungs were next homogenized with gentleMACS™ octo dissociator program m_lung_02_01, reactions were stopped with 2mM EDTA and digestions passed through 40μm cell strainers. Cells were pelleted (1500-2000rpm/300- 500xg for 5 minutes). Red blood cells were lysed with 1mL Ack lysing buffer (Lonza) for 1 minute and lysis was stopped by addition of ≥ 5 volumes of HBSS. Cells were washed once to obtain single cell suspensions. APCs were enriched with CD11c and CD11b microbeads and MACS manual separation (Miltenyi), according to the manufacturer’s instructions. Enriched cells were stained with live/dead Aqua or Via-Probe™ (BD) and with surface antibody cocktails. Populations were further purified to ≥95% purity with a BD FACSAria cell sorter. |
| Sample Type: | Sorted cells |
