Tryptophan catabolism is critical to coordinate humoral and cellular immunity to maintain hemolymph bacterial homeostasis
Description
Hemolymph samples were collected from Helicoverpa armigera fifth-instar larvae 48 h after injection with Bacillus altitudinis or PBS (control). Samples were mixed with an acetonitrile:methanol (1:1) extraction solution containing 0.02 mg/mL L-2-chlorophenylalanine, vortexed, and subjected to ultrasonic extraction. After protein precipitation, samples were centrifuged. The supernatants were dried under nitrogen, resolubilized in acetonitrile:water (1:1), and centrifuged. The clarified supernatants were analyzed using UPLC-Q-Exactive mass spectrometer (Thermo Fisher Scientific, MA, USA) with chromatographic separation on an ACQUITY HSS T3 column (Waters, Milford, USA). Mobile phases were 0.1% formic acid in water:acetonitrile (95:5) and 0.1% formic acid in water:acetonitrile:isopropanol (5:47.5:47.5). Data were processed using Progenesis QI software (Waters), and metabolite identification was performed using the HMDB database. Significant metabolites were analyzed using the “ropls” R package (version 1.6.2).
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Institutions
- Central China Normal University