Influenza A induces lactate formation to inhibit type I IFN in primary human airway epithelium. J. Thyrsted et.al. 2021
Description
Large scale metabolomics on supernatants from HPAE-ALI cells infected with either IAV or SARS-CoV-2 for 48 or 96 hours.
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For full scale metabolomics HPAE-ALI cells were harvested using pure MeOH. Medium was removed from cultures and MeOH was added and incubated for 5 min. Cells were scraped of and vortexed for 10 sec. three times. Lysed cells were centrifuged for 15 min. at 16.000 G at 4 oC. Supernatants where isolated and transported to MS-Omics on dry ice. Sample analysis was carried out by MS-Omics as follows. The analysis was carried out using a Thermo Scientific Vanquish LC coupled to Thermo Q Exactive HF MS. An electrospray ionization interface was used as ionization source. Analysis was performed in negative and positive ionization mode. The UPLC was performed using a slightly modified version of the protocol described by Catalin et al. (UPLC/MS Monitoring of Water-Soluble Vitamin Bs in Cell Culture Media in Minutes, Water Application note 2011, 720004042en). Peak areas were extracted using Compound Discoverer 2.0 (Thermo Scientific). Identification of compounds were performed at four levels; Level 1: identification by retention times (compared against in-house authentic standards), accurate mass (with an accepted deviation of 3ppm), and MS/MS spectra, Level 2a: identification by retention times (compared against in-house authentic standards), accurate mass (with an accepted deviation of 3ppm). Level 2b: identification by accurate mass (with an accepted deviation of 3ppm), and MS/MS spectra, Level 3: identification by accurate mass alone (with an accepted deviation of 3ppm).