Raw LC–MS data were processed using MS-DIAL (version X.X; Tsugawa et al., 2015) for peak detection, alignment, and normalization. Peak picking was performed with a mass accuracy tolerance of 0.01 Da for MS¹ spectra and 0.05 Da for MS² spectra. Detected features were aligned across all samples, followed by blank subtraction and filtering based on signal-to-noise ratio and feature completeness (Fill %). Internal standards were used for signal normalization and to monitor extraction and instrument performance. Metabolite and lipid identification were carried out by matching accurate mass, retention time, and MS/MS fragmentation patterns against in-house spectral libraries and public databases, including LipidBlast, MassBank, and the Human Metabolome Database (HMDB). Lipid class assignment and structural confirmation were performed following the LIPID MAPS nomenclature system and validated using characteristic MS/MS fragmentation rules for each lipid subclass. Annotation confidence levels were assigned according to the Metabolomics Standards Initiative (MSI) guidelines, with level 1 identifications supported by authentic standards and MS/MS spectral matching.
"],"repository":["MetaboLights"],"study_status":["Public"],"ptm_modification":[""],"instrument_platform":["Liquid Chromatography MS - negative - reverse phase","Liquid Chromatography MS - positive - reverse phase"],"chromatography_protocol":["Chromatographic separation of lipid species was performed on a Thermo Scientific Vanquish UHPLC system equipped with a C18 reverse-phase column (e.g., Acquity UPLC BEH C18, 2.1 × 100 mm, 1.7 µm particle size) maintained at 55 °C. The mobile phase consisted of (A) acetonitrile–water (60:40, v/v) containing 10 mM ammonium formate and 0.1 % formic acid, and (B) isopropanol–acetonitrile (90:10, v/v) containing 10 mM ammonium formate and 0.1 % formic acid. The flow rate was 0.26 mL/min, with an injection volume of 2–5 µL. A linear gradient was applied as follows: 0–2 min, 30 % B; 2–25 min, 30–100 % B; 25–30 min, 100 % B; followed by re-equilibration to initial conditions for 5 min. Samples were kept at 10 °C in the autosampler throughout the analytical sequence. The LC system was coupled online to a Q Exactive Plus Orbitrap mass spectrometer via a heated electrospray ionization (HESI) interface. Data were acquired in both positive and negative ionization modes in separate runs, ensuring optimal coverage of lipid classes.
"],"publication":["Toxoplasma gondii assembles extracellular vesicles with conserved lipid profiles across host cell types. 10.3389/fcimb.2026.1745625."],"submitter_name":["Teresa Cruz Bustos"],"submitter_affiliation":["university of veterinary medicine of Vienna"],"organism_part":["blank","Extracellular Vesicle","Cell","Quality Control"],"technology_type":["mass spectrometry assay"],"disease":[""],"extraction_protocol":["Lipid extraction was performed using a biphasic solvent system based on isopropanol (IPA) and water (90:10, v/v). For each sample, the cell or extracellular vesicle (EV) pellet was resuspended in cold extraction solvent, vortexed, and sonicated briefly to ensure complete disruption. Samples were then centrifuged to pellet debris, and the supernatant containing extracted lipids was transferred to a new tube and stored at −80 °C until analysis. Prior to LC–MS measurement, extracts were diluted appropriately in the starting mobile phase. To ensure analytical reproducibility, quality control (QC) samples were prepared by pooling equal aliquots of all biological samples and injected periodically throughout the run to monitor instrument performance and signal stability. Internal standards (IS) representing major lipid classes (e.g., deuterated LPE 18:1(d7), LPC 18:1(d7), Cer C15(d7), DG 15:0_18:1(d7), PE 15:0_18:1(d7)) were spiked into each sample before extraction to allow for normalization and relative quantification. Solvent blanks (extraction solvent without biological material) were also included to identify potential background signals and contaminants.col description
"],"organism":["Mus musculus","Toxoplasma gondii","extraction solvent (codex)","Homo sapiens","Sus scrofa","Cercopithecus","mixed sample"],"full_dataset_link":["https://www.ebi.ac.uk/metabolights/MTBLS13253"],"author":["Teresa Cruz Bustos. Postdoctoral researcher. Institute of Parasitology, Department of Biological Sciences and Pathobiology, University of Veterinary Medicine Vienna, Veterinärplatz 1, A-1210 Vienna, Austria.. teresa.cruz-bustos@vetmeduni.ac.at. +43 1 25077 2224."],"data_transformation_protocol":["Raw data files (.raw) acquired from the Thermo Q Exactive Plus Orbitrap were converted to the mzML open format using MSConvert (ProteoWizard, version X.X) with vendor peak picking enabled. The converted data were subsequently imported into MS-DIAL for spectral deconvolution, peak alignment, and normalization. Feature intensities were normalized to internal standards and total ion current (TIC) to correct for analytical variation between samples. Blanks were used for background subtraction, and features present in less than 50 % of samples within at least one group were removed. The processed dataset was exported as a tab-delimited table containing aligned peak areas for downstream statistical analysis.
Data scaling, log transformation, and imputation of missing values (KNN method) were carried out in MetaboAnalyst and R to improve data normality and comparability across samples.
"],"study_factor":["Origin"],"submitter_email":["teresa.cruz-bustos@vetmeduni.ac.at"],"sample_collection_protocol":["The procedures for Toxoplasma gondii culture, extracellular vesicle (EV) isolation and characterisation were conducted as previously described (Cruz-Bustos et al. 2025). In brief, RH strain tachyzoites were propagated in four different host cell lines, human foreskin fibroblasts (Hs27, obtained from ATCC CRL-1634), green monkey kidney epithelial cells (Vero, ATCC CCL-181), mouse myoblasts (C2C12, ATCC CRL-1771) and porcine intestinal epithelial cells (IPEC-1, ACC 705, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures GmbH, Leibniz, Germany), to isolate EVs produced in diverse cellular environments. Following egress, the parasites were incubated in EV-depleted medium under host cell-free conditions to allow EV release. The EVs were then isolated from the conditioned medium via a series of differential centrifugation steps, including 0.22 µm filtration and ultracentrifugation at 100,000×g. The purity and morphology of the EVs were subsequently assessed using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM), in accordance with the MISEV2023 guidelines. For the present study, only TgEVs derived from tachyzoites and whole cells were used for lipidomic profiling.
"],"omics_type":["Metabolomics"],"study_design":["Toxoplasma gondii","Lipidome","Extracellular Vesicles"],"curator_keywords":["Toxoplasma gondii","Lipidome","Extracellular Vesicles"],"mass_spectrometry_protocol":["Mass spectrometric analysis was performed using a Q Exactive Plus Orbitrap mass spectrometer (Thermo Fisher Scientific, Bremen, Germany) equipped with a heated electrospray ionization (HESI) source. Data were acquired in both positive and negative ionization modes in separate runs, covering an m/z range of 200–2000. The spray voltage was set to 3.5 kV in positive mode and 2.5 kV in negative mode, with a capillary temperature of 320 °C. The sheath gas and auxiliary gas were maintained at 40 and 10 arbitrary units, respectively, and the S-lens RF level was set to 50. Full MS scans were acquired at a resolution of 70,000 (at m/z 200), with an automatic gain control (AGC) target of 1 × 1000000 ions and a maximum injection time of 100 ms, resulting in an approximate scan rate of 2 Hz. Data-dependent MS/MS acquisition was performed for the top 10 most intense precursor ions per scan using higher-energy collisional dissociation (HCD) at a normalized collision energy (NCE) of 25–30 eV. Dynamic exclusion was enabled to prevent repeated fragmentation of the same ion species. Instrument calibration was performed daily using Thermo Fisher calibration solutions to ensure mass accuracy within 5 ppm.
"],"metabolite_name":["TG O-62:6|TG O-22:0_18:1_22:5","PC 31:1","TG 67:4|TG 18:1_25:1_24:2","PC O-40:2|PC O-16:0_24:2","HexCer 46:1;O2|HexCer 20:1;O2/26:0","PC 42:3|PC 18:1_24:2","PE P-41:4|PE P-17:0_24:4","PE 33:2|PE 16:1_17:1","TG O-60:3|TG O-18:1_20:1_22:1","TG 63:1|TG 18:0_27:0_18:1","SM 31:1;O2","TG 57:5|TG 17:0_18:0_22:5","Cer 42:1;O2|Cer 18:1;O2/24:0","PE P-37:5|PE P-15:0_22:5","LPE 24:0","Hex3Cer 42:3;O2|Hex3Cer 18:1;O2/24:2","PE 32:2|PE 16:1_16:1","PC O-34:3|PC O-16:1_18:2","LPE 24:1","LPE 24:2","PE 38:5","PE 38:4","PE 38:3","HexCer 35:1;O2|HexCer 18:1;O2/17:0","PE 38:2","SM 40:3;O2","CL 67:4|CL 33:2_34:2","SPB 20:0;O2","TG 56:0|TG 16:0_16:0_24:0","PC 44:5|PC 24:1_20:4","BMP 44:10|BMP 22:4_22:6","PE 38:7","SM 59:2;O3|SM 41:1;O2(FA 18:0)","TG O-56:2|TG O-20:0_18:1_18:1","TG 47:0|TG 15:0_16:0_16:0","TG O-58:7|TG O-22:5_18:1_18:1","DG 46:1|DG 20:0_26:1","LPC 26:0/0:0","TG 68:10|TG 24:1_22:4_22:5","SM 44:3;O2|SM 18:1;O2/26:2","TG 66:10|TG 18:0_22:5_26:5","PE P-35:1|PE P-17:0_18:1","PC 44:12|PC 22:6_22:6","PE P-40:4|PE P-18:0_22:4","SM 41:5;O2","TG 71:4|TG 18:1_27:1_26:2","TG O-60:5|TG O-22:0_16:0_22:5","PI 37:5","PC O-34:6|PC O-12:0_22:6","PI 37:4","TG O-54:6|TG O-22:6_16:0_16:0","PC O-39:7|PC O-17:1_22:6","PI 37:3","TG 28:0|TG 8:0_10:0_10:0","HexCer 40:2;O2|HexCer 18:2;O2/22:0","CL 66:2|CL 32:1_34:1","PI 37:2","TG O-62:3|TG O-18:1_22:1_22:1","CL 66:3|CL 32:1_34:2","TG 66:9|TG 18:1_26:2_22:6","Cer 45:1;O2|Cer 20:1;O2/25:0","TG O-60:7|TG O-22:1_16:0_22:6","CL 69:5|CL 33:2_36:3","PC O-31:0|PC O-16:0_15:0","TG 60:1|TG 16:0_26:0_18:1","TG 61:2|TG 17:0_18:1_26:1","PC 40:6|PC 18:0_22:6","HBMP 58:8|HBMP 18:1/18:1_22:6","Cer 36:2;O2|Cer 18:2;O2/18:0","CL 70:8|CL 32:2_38:6","TG 63:3|TG 18:1_18:1_27:1","HexCer 38:0;O2|HexCer 18:0;O2/20:0","TG 62:9|TG 18:0_22:4_22:5","TG 58:5|TG 18:0_18:1_22:4","DG 32:1|DG 16:0_16:1","CL 68:4|CL 32:1_36:3","SM 46:3;O2|SM 18:1;O2/28:2","TG 52:2|TG 16:0_18:1_18:1","PC 42:1","PC 42:2","PC 42:0","TG 72:2|TG 26:0_20:1_26:1","PC 42:3","Cer 44:4;O2|Cer 18:1;O2/26:3","TG O-58:5|TG O-18:0_18:0_22:5","PC 42:4","PC 43:1|PC 17:0_26:1","Hex2Cer 46:1;O2|Hex2Cer 18:1;O2/28:0","CL 69:6|CL 34:3_35:3","TG 48:2|TG 14:0_16:1_18:1","PC 44:2|PC 18:1_26:1","TG 59:2|TG 16:0_18:1_25:1","TG 51:6|TG 14:0_15:0_22:6","TG 65:4|TG 18:1_26:1_21:2","PE 43:1|PE 25:0_18:1","LPC 21:0/0:0","PE 34:2|PE 16:1_18:1","PE P-31:1|PE P-15:0_16:1","PC 36:0|PC 18:0_18:0","TG 40:0|TG 10:0_14:0_16:0","Hex3Cer 36:1;O2|Hex3Cer 18:1;O2/18:0","TG O-54:4|TG O-18:2_18:1_18:1","DG 36:0|DG 16:0_20:0","TG 66:14|TG 22:4_22:5_22:5","Cer 38:1;O2|Cer 18:1;O2/20:0","PC O-37:2|PC O-16:0_21:2","SM 35:2;O2|SM 18:2;O2/17:0","PE 38:5|PE 16:0_22:5","PC O-41:0","PC O-41:1","TG 62:10|TG 18:0_22:5_22:5","TG 59:5|TG 16:0_21:1_22:4","TG 60:8|TG 18:1_20:2_22:5","CE 30:2","SM 32:1;O2|SM 18:1;O2/14:0","PC 46:2|PC 18:1_28:1","Cer 44:3;O2|Cer 18:1;O2/26:2","PC O-38:8|PC O-16:2_22:6","CE 30:5","CE 30:6","CE 30:3","PC O-34:1|PC O-16:0_18:1","CE 30:4","LPE 22:0","LPE 22:1","LPE 22:2","PE P-40:6|PE P-18:0_22:6","DG 32:0|DG 16:0_16:0","PC O-30:3","TG O-58:1|TG O-18:0_20:0_20:1","Cer 36:0;O2|Cer 18:0;O2/18:0","TG 48:0|TG 14:0_16:0_18:0","Cer 43:3;O2|Cer 18:2;O2/25:1","TG O-52:1|TG O-16:0_16:0_20:1","PC 40:0","CAR 18:2","CAR 18:1","DG 38:1|DG 16:0_22:1","CAR 18:0","PC 28:0|PC 14:0_14:0","TG 54:2|TG 16:0_18:1_20:1","DG 38:0|DG 16:0_22:0","TG 62:0|TG 16:0_22:0_24:0","CAR 18:3","Cer 48:2;O2|Cer 18:1;O2/30:1","SM 34:0;O2","HexCer 38:2;O2|HexCer 18:1;O2/20:1","PE P-40:2|PE P-18:0_22:2","DG 39:5|DG 17:0_22:5","TG 61:6|TG 17:0_18:1_26:5","LPC 17:1/0:0","TG 59:0|TG 16:0_18:0_25:0","PE P-41:2|PE P-17:0_24:2","SM 40:3;O2|SM 18:1;O2/22:2","PI 35:3","TG 60:0|TG 16:0_18:0_26:0","HBMP 58:7|HBMP 18:1/18:0_22:6","HexCer 44:0;O2|HexCer 20:0;O2/24:0","PC 42:0|PC 16:0_26:0","PI 35:2","PI 35:1","TG O-52:3|TG O-18:1_16:1_18:1","TG 55:1|TG 16:0_21:0_18:1","TG 61:0|TG 16:0_20:0_25:0","TG 64:3|TG 18:1_20:1_26:1","PC O-39:5|PC O-17:0_22:5","PE P-38:4|PE P-18:0_20:4","TG 62:4|TG 18:1_18:1_26:2","DG 35:2|DG 17:1_18:1","PE P-35:3|PE P-16:0_19:3","TG O-56:4|TG O-16:0_18:1_22:3","SM 39:1;O2","TG O-52:2|TG O-18:1_16:0_18:1","TG O-61:2|TG O-22:0_17:1_22:1","PC O-35:3|PC O-16:2_19:1","DG 37:4|DG 17:0_20:4","PE P-37:3|PE P-16:0_21:3","PC O-33:4|PC O-13:0_20:4","TG 55:7|TG 16:0_17:1_22:6","TG O-46:1|TG O-18:1_14:0_14:0","DG 42:6|DG 16:0_26:6","TG O-56:5|TG O-18:0_16:0_22:5","PC O-27:0|PC O-11:0_16:0","PC O-40:5|PC O-18:0_22:5","HBMP 54:1|HBMP 18:1/18:0_18:0","PC 40:3","PC 40:4","BMP 42:8|BMP 20:2_22:6","SM 42:4;O2|SM 18:1;O2/24:3","PC 40:1","PC 40:2","TG 62:7|TG 20:1_20:1_22:5","PC 40:6","TG 71:2|TG 26:0_18:1_27:1","Hex3Cer 42:2;O2|Hex3Cer 18:1;O2/24:1","DG 52:3|DG 26:1_26:2","TG 56:8|TG 16:1_18:1_22:6","SM 54:2;O3|SM 36:1;O2(FA 18:0)","DG 34:1|DG 16:0_18:1","PE 34:3|PE 16:1_18:2","PE P-38:0|PE P-18:0_20:0","PC O-43:1","PC O-43:6","TG 68:2|TG 16:0_26:1_26:1","PC O-43:7","PC O-43:4","SM 34:1;O2|SM 18:1;O2/16:0","SM 43:0;O2","BMP 34:1|BMP 16:0_18:1","PI 33:1","CE 32:5","CE 32:6","TG 74:8|TG 26:1_26:1_22:6","TG 48:0|TG 16:0_16:0_16:0","PC 40:1|PC 16:0_24:1","TG 63:3|TG 18:1_19:1_26:1","TG 66:13|TG 18:1_22:6_26:6","Hex3Cer 41:1;O2|Hex3Cer 17:1;O2/24:0","TG 57:8|TG 17:1_18:1_22:6","CE 32:4","SM 45:0;O2|SM 18:0;O2/27:0","PC O-37:0|PC O-21:0_16:0","Hex2Cer 43:2;O2|Hex2Cer 19:1;O2/24:1","LPE 20:1","SM 34:2;O2","TG O-58:6|TG O-22:4_18:1_18:1","LPE 20:4","DG 48:3|DG 22:1_26:2","PE 32:0|PE 14:0_18:0","TG 65:8|TG 18:1_25:1_22:6","LPE 20:0","TG 57:4|TG 18:0_18:1_21:3","PC 34:5|PC 14:0_20:5","TG 74:3|TG 22:1_24:1_28:1","TG O-58:2|TG O-16:0_20:1_22:1","TG O-48:0|TG O-16:0_16:0_16:0","CAR 16:1","TG O-58:5|TG O-22:5_18:0_18:0","PC 46:6|PC 24:0_22:6","TG 64:8|TG 18:1_24:1_22:6","PE P-38:6|PE P-16:0_22:6","TG 56:5|TG 16:0_18:0_22:5","CAR 16:0","SM 38:2;O2|SM 18:2;O2/20:0","TG 51:5|TG 15:0_16:0_20:5","TG 53:4|TG 17:1_18:1_18:2","LPC O-21:0","TG 62:6|TG 16:1_24:1_22:4","BMP 38:5|BMP 18:1_20:4","SM 45:2;O2|SM 18:1;O2/27:1","PC 42:8|PC 20:4_22:4","PC O-38:2|PC O-16:0_22:2","PE P-39:4|PE P-17:0_22:4","TG O-62:7|TG O-22:1_18:1_22:5","TG O-54:3|TG O-18:1_18:1_18:1","DG O-42:5|DG O-18:0_24:5","HexCer 40:1;O3|HexCer 18:1;O2/22:0;O","PC 50:6|PC 28:1_22:5","PC 40:4|PC 18:0_22:4","TG 60:4|TG 18:1_24:1_18:2","BMP 38:4|BMP 18:1_20:3","BMP 44:11|BMP 22:5_22:6","PC 44:9|PC 22:4_22:5","TG 59:1|TG 16:0_25:0_18:1","PC O-43:5|PC O-23:1_20:4","Cer 45:2;O2|Cer 18:1;O2/27:1","DG 32:2|DG 16:1_16:1","PE P-35:4|PE P-15:0_20:4","TG O-54:5|TG O-22:5_16:0_16:0","CE 21:5","PE P-42:5|PE P-18:0_24:5","PE 33:1|PE 16:0_17:1","PE P-36:3|PE P-16:0_20:3","PC O-40:4|PC O-16:0_24:4","TG 66:1|TG 16:0_24:0_26:1","CL 66:4|CL 32:2_34:2","TG 53:3|TG 16:1_18:1_19:1","TG O-58:8|TG O-22:6_18:1_18:1","CE 21:3","DG 37:3|DG 16:0_21:3","TG 60:2|TG 16:0_18:1_26:1","CE 21:4","CL 76:7|CL 38:3_38:4","SM 32:2;O2|SM 18:2;O2/14:0","TG 65:2|TG 16:0_24:1_25:1","CL 68:5|CL 34:2_34:3","TG 59:4|TG 18:1_19:1_22:2","TG 70:8|TG 24:1_24:1_22:6","Cer 44:2;O2|Cer 18:2;O2/26:0","TG O-64:10|TG O-20:0_22:5_22:5","TG 66:6|TG 18:0_26:1_22:5","Cer 46:0;O2|Cer 20:0;O2/26:0","TG 49:0|TG 15:0_16:0_18:0","TG 60:3|TG 16:1_18:1_26:1","TG 50:5|TG 14:0_16:1_20:4","LPC 20:2/0:0","PC 42:7|PC 18:1_24:6","CE 34:6","DG 38:2|DG 18:1_20:1","TG 76:4|TG 24:1_26:1_26:2","TG 67:2|TG 16:0_25:1_26:1","TG 72:3|TG 18:1_26:1_28:1","PE 40:7","HexCer 42:2;O2|HexCer 18:1;O2/24:1","PC O-28:1|PC O-14:1_14:0","PC O-35:5|PC O-15:1_20:4","PC 40:5|PC 18:0_22:5","PE 40:5","PE 40:6","DG 39:2|DG 18:1_21:1","TG 62:6|TG 18:0_18:1_26:5","SM 44:2;O2","Cer 41:0;O2|Cer 18:0;O2/23:0","DG 39:1|DG 16:0_23:1","TG O-57:7|TG O-17:0_18:1_22:6","Cer 34:1;O2|Cer 18:1;O2/16:0","HexCer 41:1;O2|HexCer 18:1;O2/23:0","TG 50:3|TG 16:0_16:1_18:2","SM 50:2;O3|SM 34:1;O2(FA 16:0)","TG 67:6|TG 20:1_25:1_22:4","TG 61:3|TG 18:1_18:1_25:1","DG 35:1|DG 16:0_19:1","PC O-46:5|PC O-24:1_22:4","TG 54:9|TG 16:0_16:3_22:6","TG O-54:0|TG O-16:0_18:0_20:0","TG O-54:5|TG O-16:0_16:0_22:5","Cer 54:3;O3|Cer 18:1;O2/36:2;O","CAR 14:1","TG 61:9|TG 18:1_21:3_22:5","CAR 14:0","BMP 40:8|BMP 18:2_22:6","TG O-52:5|TG O-16:0_14:0_22:5","TG 66:5|TG 18:1_26:1_22:3","TG O-47:0|TG O-17:0_15:0_15:0","Dioctyl phthalate (also known as the production of plastic)","Cer 41:0;O2|Cer 17:0;O2/24:0","PE 34:1|PE 16:0_18:1","TG 43:1|TG 13:0_14:0_16:1","PE 34:0|PE 16:0_18:0","CL 67:5|CL 33:2_34:3","TG O-54:0|TG O-22:0_16:0_16:0","CL 69:4|CL 34:2_35:2","TG 64:1|TG 16:0_24:0_24:1","PC 48:8|PC 26:2_22:6","PC O-36:2|PC O-18:1_18:1","PC 36:2|PC 18:1_18:1","CoQ10","TG 72:9|TG 24:1_26:2_22:6","TG 63:4|TG 18:1_26:1_19:2","TG 48:4|TG 16:0_16:1_16:3","TG 58:3|TG 18:1_20:1_20:1","PC O-33:1|PC O-17:1_16:0","PC O-28:2|PC O-16:2_12:0","DG 42:4|DG 16:0_26:4","SM 41:2;O2|SM 18:1;O2/23:1","PC 44:6|PC 24:1_20:5","PC 44:3|PC 18:1_26:2","SM 43:3;O2|SM 17:1;O2/26:2","TG O-48:3|TG O-18:1_14:1_16:1","TG 64:5|TG 18:1_26:1_20:3","PE P-38:2|PE P-18:0_20:2","PC O-36:7|PC O-14:1_22:6","PC O-37:6|PC O-15:1_22:5","CL 74:4|CL 36:1_38:3","PC 48:2|PC 18:1_30:1","TG O-58:7|TG O-18:0_18:1_22:6","SM 36:2;O2|SM 18:2;O2/18:0","CL 66:5|CL 32:2_34:3","LPC 20:3/0:0","SM 53:2;O3|SM 25:1;O2(FA 28:0)","TG 54:4|TG 16:0_18:1_20:3","PE P-37:1|PE P-16:0_21:1","TG 55:2|TG 16:0_19:1_20:1","TG 50:3|TG 16:1_16:1_18:1","TG 54:0|TG 16:0_18:0_20:0","TG 45:2|TG 14:0_15:1_16:1","TG 53:1|TG 17:0_18:0_18:1","PC 42:6|PC 18:1_24:5","TG 64:10|TG 16:0_22:5_26:5","HexCer 44:2;O3|HexCer 18:1;O2/26:1;O","TG 68:5|TG 18:1_26:1_24:3","ST 27:1;O","TG O-58:2|TG O-22:0_18:1_18:1","SM 40:1;O3|SM 24:0;O2(FA 16:0)","TG 65:6|TG 17:0_26:1_22:5","CE 36:6","HexCer 41:0;O2|HexCer 18:0;O2/23:0","PE 42:6","DG 44:1|DG 16:0_28:1","PE 42:1","DG 40:6|DG 18:1_22:5","PE 42:2","TG 57:0|TG 16:0_17:0_24:0","VAE 27:1","HexCer 44:1;O2|HexCer 20:1;O2/24:0","TG O-48:1|TG O-16:0_16:0_16:1","TG 68:6|TG 18:1_26:1_24:4","TG 48:3|TG 16:1_16:1_16:1","CL 74:5|CL 36:2_38:3","Hex2Cer 42:1;O2|Hex2Cer 20:1;O2/22:0","PI 42:10","PC O-44:7|PC O-22:1_22:6","DG 30:0|DG 14:0_16:0","TG O-57:3|TG O-17:0_18:1_22:2","PE P-42:6|PE P-18:1_24:5","TG 62:1|TG 18:0_26:0_18:1","HBMP 56:3|HBMP 18:1/18:1_20:1","SM 40:2;O2|SM 18:1;O2/22:1","DG 40:1|DG 16:0_24:1","PE P-42:2|PE P-18:0_24:2","Hex3Cer 41:2;O2|Hex3Cer 17:1;O2/24:1","LPC 14:0/0:0","PC O-32:2|PC O-16:0_16:2","PE P-34:3|PE P-16:0_18:3","TG 66:4|TG 26:0_18:1_22:3","TG O-51:2|TG O-17:1_16:0_18:1","PI 30:1","TG 59:11|TG 15:0_22:5_22:6","TG 64:2|TG 16:0_24:1_24:1","Hex2Cer 34:1;O2|Hex2Cer 18:1;O2/16:0","PC 38:2","PC 38:3","BMP 38:6|BMP 16:0_22:6","PC 38:1","PC 38:6","PC 38:7","TG 53:0|TG 16:0_18:0_19:0","PC 38:4","PC O-26:0|PC O-10:0_16:0","TG O-50:1|TG O-18:1_16:0_16:0","PC 38:5","TG 57:6|TG 18:1_19:1_20:4","DG O-32:0|DG O-16:0_16:0","SM 58:2;O3|SM 40:1;O2(FA 18:0)","PC O-34:4|PC O-14:0_20:4","LPC O-15:1","LPC O-15:0","PE P-36:2|PE P-18:1_18:1","CE 25:1","TG O-60:10|TG O-16:0_22:5_22:5","Cer 41:2;O2|Cer 18:1;O2/23:1","PC 48:7|PC 26:1_22:6","TG 60:4|TG 18:1_18:1_24:2","TG 56:6|TG 16:0_18:1_22:5","BMP 38:3|BMP 18:1_20:2","HexCer 42:2;O3|HexCer 18:1;O2/24:1;O","PE 40:0","PC O-36:6|PC O-16:1_20:5","PE 40:1","PE 40:2","Hex3Cer 38:1;O2|Hex3Cer 20:1;O2/18:0","Cer 44:0;O3|Cer 18:0;O2/26:0;O","TG 54:0|TG 14:0_16:0_24:0","CL 72:5|CL 36:2_36:3","TG O-58:9|TG O-16:0_20:4_22:5","LPC O-26:1","CL 71:2|CL 35:1_36:1","PC O-44:5|PC O-22:0_22:5","PE P-36:5|PE P-14:0_22:5","TG O-56:7|TG O-16:0_18:1_22:6","TG 44:1|TG 12:0_16:0_16:1","TG 68:9|TG 26:1_20:3_22:5","Cer 48:4;O2|Cer 18:1;O2/30:3","PE 44:4","TG O-55:6|TG O-16:0_17:1_22:5","PC 40:10|PC 20:5_20:5","PE 44:5","TG 53:6|TG 15:0_16:0_22:6","DG 48:5|DG 26:0_22:5","TG 66:16|TG 22:5_22:5_22:6","DG O-38:5|DG O-16:0_22:5","PE 44:1","PE 44:2","PC 35:4|PC 15:0_20:4","Cer 39:0;O2|Cer 17:0;O2/22:0","BMP 42:9|BMP 20:3_22:6","HBMP 53:3|HBMP 18:1/17:1_18:1","SM 33:0;O2","SM 44:5;O2|SM 18:1;O2/26:4","PC O-32:0|PC O-16:0_16:0","TG 60:9|TG 16:0_22:4_22:5","TG O-52:4|TG O-18:1_16:1_18:2","TG O-54:6|TG O-16:0_16:1_22:5","CL 64:2|CL 30:1_34:1","CL 65:4|CL 32:2_33:2","TG O-55:0|TG O-16:0_18:0_21:0","HBMP 52:3|HBMP 18:1/16:1_18:1","PC O-31:2|PC O-15:1_16:1","TG 60:13|TG 16:1_22:6_22:6","TG 57:2|TG 16:0_18:1_23:1","HexCer 32:2;O2|HexCer 18:2;O2/14:0","TG 68:2|TG 24:0_18:1_26:1","DG 39:4|DG 19:0_20:4","SM 41:1;O2","HexCer 41:2;O2|HexCer 18:1;O2/23:1","Cer 44:2;O2|Cer 18:1;O2/26:1","DG 36:2|DG 18:1_18:1","PE 36:5|PE 16:1_20:4","TG 62:12|TG 18:1_22:5_22:6","TG O-60:9|TG O-16:0_22:4_22:5","Hex2Cer 41:2;O2|Hex2Cer 17:1;O2/24:1","TG O-48:2|TG O-14:0_16:1_18:1","TG 70:9|TG 26:1_22:2_22:6","PE P-36:1|PE P-18:0_18:1","CE 16:0","CE 16:1","TG O-56:7|TG O-20:5_18:1_18:1","PC 36:1","PC 36:4","PC 39:4|PC 18:1_21:3","CL 67:3|CL 33:1_34:2","PC 36:5","SM 42:1;O2","PC 36:2","PC 36:3","TG 45:0|TG 14:0_15:0_16:0","PE P-36:6|PE P-14:0_22:6","PC 36:6","LPC 16:1/0:0","PC O-46:6|PC O-24:1_22:5","SM 44:4;O2|SM 18:1;O2/26:3","SM 56:3;O3|SM 40:2;O2(FA 16:0)","TG 46:2|TG 14:0_16:1_16:1","Hex3Cer 34:1;O2|Hex3Cer 18:1;O2/16:0","PE 38:6|PE 16:1_22:5","TG 69:4|TG 18:1_25:1_26:2","SM 58:3;O3|SM 42:2;O2(FA 16:0)","CE 27:1","SM 32:1;O2","PC O-38:7|PC O-18:2_20:5","PE 41:1|PE 23:0_18:1","TG O-54:7|TG O-22:5_16:1_16:1","PC 36:5|PC 16:1_20:4","TG O-47:1|TG O-17:1_14:0_16:0","SM 60:4;O3|SM 42:2;O2(FA 18:1)","Hex3Cer 44:3;O2|Hex3Cer 18:2;O2/26:1","PC O-42:6|PC O-20:0_22:6","LPC O-24:0","SM 56:2;O3|SM 40:1;O2(FA 16:0)","LPC 18:2/0:0","TG O-56:6|TG O-16:0_18:1_22:5","Hex3Cer 33:1;O2|Hex3Cer 18:1;O2/15:0","SM 46:1;O2|SM 20:1;O2/26:0","DG O-34:1|DG O-18:1_16:0","PC O-36:5|PC O-16:1_20:4","DG 46:4|DG 26:1_20:3","PC O-38:3|PC O-18:1_20:2","LPC O-24:1","Cer 48:1;O2|Cer 20:0;O2/28:1","TG O-57:2|TG O-17:0_18:1_22:1","TG 55:0|TG 14:0_16:0_25:0","HexCer 42:0;O2|HexCer 18:0;O2/24:0","TG O-56:5|TG O-20:5_18:0_18:0","PE 46:5","DG 34:2|DG 16:1_18:1","PE 46:6","LPC 18:1/0:0","PC O-46:7|PC O-24:1_22:6","PE 46:7","PC 37:4|PC 17:0_20:4","PE 46:2","PE 30:0|PE 14:0_16:0","HexCer 42:0;O4|HexCer 18:0;O3/24:0(2OH)","PE 38:1|PE 18:0_20:1","HexCer 42:1;O4|HexCer 18:0;O3/24:1(2OH)","PC O-32:5|PC O-12:0_20:5","SM 37:0;O2","TG 50:4|TG 16:1_18:1_16:2","HexCer 42:3;O2|HexCer 18:2;O2/24:1","DG 48:7|DG 26:1_22:6","PC O-38:5|PC O-18:1_20:4","HBMP 56:4|HBMP 18:1/18:1_20:2","SM 38:3;O2","PC O-35:1|PC O-17:0_18:1","PE 42:7|PE 18:1_24:6","SM 41:1;O2|SM 18:1;O2/23:0","SM 36:1;O2|SM 18:1;O2/18:0","TG 52:5|TG 16:0_16:1_20:4","PC 42:1|PC 16:0_26:1","TG O-55:3|TG O-18:0_16:0_21:3","TG 64:6|TG 16:0_26:1_22:5","SPB 18:0;O2","TG 66:12|TG 18:1_26:5_22:6","TG 71:3|TG 18:1_26:1_27:1","Cer 46:3;O2|Cer 18:1;O2/28:2","SM 52:3;O3|SM 34:1;O2(FA 18:1)","TG 61:1|TG 17:0_26:0_18:1","CL 70:3|CL 34:1_36:2","PC 35:1|PC 17:0_18:1","TG 58:10|TG 16:0_20:4_22:6","PC O-41:4|PC O-21:0_20:4","PC O-36:4|PC O-16:0_20:4","Cer 40:3;O2|Cer 18:2;O2/22:1","LPE 17:0","SM 43:4;O2","CE 18:0","PC 34:2","CE 18:2","TG 69:3|TG 18:1_25:1_26:1","PC 34:3","CE 18:1","PC 34:0","Cer 34:0;O2|Cer 18:0;O2/16:0","SM 41:3;O2|SM 18:2;O2/23:1","PC 34:1","TG 52:4|TG 16:0_16:1_20:3","TG 63:0|TG 16:0_23:0_24:0","PC 46:4|PC 26:1_20:3","TG 63:6|TG 16:0_25:1_22:5","PC 34:4","LPC 17:0/0:0","PC 34:5","Cer 32:1;O2|Cer 18:1;O2/14:0","TG 46:0|TG 14:0_16:0_16:0","PC O-38:0|PC O-18:0_20:0","TG 68:3|TG 18:1_24:1_26:1","LPC O-19:1","LPC O-19:0","DG 38:3|DG 18:0_20:3","HexCer 32:1;O2|HexCer 18:1;O2/14:0","HexCer 39:1;O2|HexCer 17:1;O2/22:0","TG O-62:10|TG O-18:0_22:5_22:5","PE P-44:5|PE P-24:1_20:4","TG 54:6|TG 16:0_16:0_22:6","TG 60:8|TG 18:1_20:3_22:4","PE P-38:1|PE P-16:0_22:1","TG O-55:2|TG O-16:0_18:1_21:1","Hex2Cer 34:2;O2|Hex2Cer 18:1;O2/16:1","TG O-53:1|TG O-19:0_16:0_18:1","TG O-48:4|TG O-12:0_16:0_20:4","LPC 16:0/0:0","DG O-40:8|DG O-18:2_22:6","LPC 19:1/0:0","CL 65:3|CL 31:1_34:2","DG 42:2|DG 18:1_24:1","DG 34:0|DG 16:0_18:0","Cer 43:0;O2|Cer 18:0;O2/25:0","SM 55:2;O3|SM 39:1;O2(FA 16:0)","TG 64:9|TG 16:0_22:4_26:5","TG O-60:6|TG O-22:1_16:0_22:5","HexCer 40:0;O2|HexCer 18:0;O2/22:0","TG O-51:1|TG O-18:1_16:0_17:0","Cer 41:1;O2|Cer 18:1;O2/23:0","TG 63:7|TG 16:1_25:1_22:5","TG O-57:6|TG O-17:0_18:1_22:5","TG 68:4|TG 16:1_26:1_26:2","PE 48:7","Cer 42:4;O2|Cer 18:2;O2/24:2","PC 46:7|PC 24:1_22:6","BMP 44:12|BMP 22:6_22:6","PE 48:6","TG 61:10|TG 18:1_21:3_22:6","TG 62:10|TG 18:1_22:4_22:5","CL 72:3|CL 36:1_36:2","SM 43:2;O2|SM 18:1;O2/25:1","SM 57:3;O3|SM 41:2;O2(FA 16:0)","ST 28:1;O","TG O-62:1|TG O-18:0_22:0_22:1","LPC 18:0/0:0","PC O-31:1|PC O-16:1_15:0","CL 69:3|CL 34:1_35:2","PC O-41:5|PC O-17:0_24:5","TG 60:12|TG 16:0_22:6_22:6","DG 42:5|DG 18:0_24:5","PC 38:3|PC 18:0_20:3","TG O-50:5|TG O-12:0_16:0_22:5","TG O-62:8|TG O-18:0_22:4_22:4","BMP 40:5|BMP 18:1_22:4","Hex3Cer 42:1;O2|Hex3Cer 20:1;O2/22:0","TG 70:7|TG 18:1_26:1_26:5","DG O-40:3|DG O-18:1_22:2","LPE 26:1","HexCer 43:1;O2|HexCer 18:1;O2/25:0","SM 34:2;O2|SM 18:2;O2/16:0","TG O-54:7|TG O-14:0_18:1_22:6","TG 55:3|TG 18:1_18:1_19:1","Cer 40:1;O2|Cer 18:1;O2/22:0","PE 39:2|PE 18:1_21:1","Cer 35:0;O2|Cer 18:0;O2/17:0","TG O-60:1|TG O-18:0_20:0_22:1","PE 45:2|PE 18:1_27:1","TG O-60:8|TG O-18:0_20:4_22:4","SM 52:4;O3|SM 34:2;O2(FA 18:1)","PI 39:6","PI 39:5","HexCer 34:2;O2|HexCer 18:2;O2/16:0","PI 39:4","PI 39:3","TG 45:1|TG 14:0_15:0_16:1","TG 58:1|TG 16:0_24:0_18:1","PE P-42:7|PE P-18:1_24:6","SM 43:2;O2","PC 32:0","PC 32:1","Cer 43:2;O2|Cer 18:1;O2/25:1","PC 42:2|PC 18:1_24:1","DG 33:1|DG 16:0_17:1","TG 46:3|TG 14:1_16:1_16:1","PC 32:2","PC 32:3","PC 34:0|PC 16:0_18:0","TG 55:8|TG 16:1_17:1_22:6","CL 68:6|CL 32:2_36:4","TG O-56:0|TG O-16:0_18:0_22:0","Cer 50:5;O2|Cer 18:1;O2/32:4","TG O-52:7|TG O-14:0_16:1_22:6","TG 69:2|TG 25:0_18:1_26:1","PE P-34:1|PE P-16:0_18:1","BMP 34:2|BMP 16:1_18:1","TG O-56:3|TG O-20:1_18:1_18:1","Cer 44:0;O2|Cer 20:0;O2/24:0","DG 38:6|DG 16:0_22:6","TG 55:3|TG 18:0_18:1_19:2","LPC O-17:1","PC 44:7|PC 18:1_26:6","LPC O-17:0","Cer 50:2;O3|Cer 18:1;O2/32:1;O","BMP 40:7|BMP 18:1_22:6","TG 70:6|TG 24:0_24:1_22:5","TG O-53:2|TG O-18:1_16:0_19:1","TG O-44:2|TG O-8:0_18:1_18:1","TG 36:0|TG 10:0_12:0_14:0","PE 36:2|PE 18:1_18:1","Hex3Cer 34:2;O2|Hex3Cer 18:1;O2/16:1","TG O-60:4|TG O-18:1_20:1_22:2","CL 70:7|CL 34:2_36:5","PC O-44:6|PC O-22:0_22:6","Hex2Cer 35:1;O2|Hex2Cer 19:1;O2/16:0","TG 62:8|TG 16:1_24:1_22:6","TG 62:4|TG 18:1_26:1_18:2","Cer 42:0;O3|Cer 18:0;O2/24:0;O","TG 58:4|TG 18:0_18:1_22:3","TG O-58:6|TG O-18:0_18:1_22:5","TG 55:1|TG 18:0_19:0_18:1","TG 55:5|TG 16:0_17:0_22:5","TG 68:4|TG 18:1_24:1_26:2","TG 56:7|TG 16:0_18:1_22:6","LPC 19:0/0:0","PE 36:1|PE 18:0_18:1","LPC 28:1/0:0","PE P-33:1|PE P-16:0_17:1","SM 38:0;O2","PC O-42:2|PC O-18:1_24:1","TG O-45:0|TG O-13:0_16:0_16:0","Hex3Cer 44:1;O2|Hex3Cer 20:1;O2/24:0","CL 74:9|CL 36:3_38:6","PC O-30:2|PC O-14:1_16:1","TG 57:9|TG 17:1_18:2_22:6","PE 44:3|PE 18:1_26:2","TG O-49:0|TG O-16:0_16:0_17:0","CL 64:4|CL 30:2_34:2","TG 58:0|TG 16:0_18:0_24:0","TG O-50:1|TG O-16:0_16:0_18:1","PC 34:3|PC 16:1_18:2","CL 68:1|CL 34:0_34:1","PC O-39:4|PC O-18:1_21:3","HexCer 43:0;O2|HexCer 18:0;O2/25:0","TG 65:5|TG 18:1_26:1_21:3","PC O-38:4|PC O-18:0_20:4","TG 58:6|TG 18:0_18:1_22:5","PC 43:1","TG 66:4|TG 16:1_26:1_24:2","Cer 38:2;O2|Cer 18:2;O2/20:0","PC 43:2","TG 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16:0_18:0_18:0","PC 46:4","TG 74:6|TG 26:1_26:1_22:4","TG O-54:2|TG O-14:0_20:1_20:1","Cer 37:1;O2|Cer 18:1;O2/19:0","PC 46:7","TG O-52:0|TG O-16:0_18:0_18:0","DG 37:1|DG 16:0_21:1","PC O-37:5|PC O-17:1_20:4","TG O-62:2|TG O-18:0_22:1_22:1","LPE 18:0","LPE 18:1","PC O-41:2","PC O-41:3","PC O-44:8|PC O-22:2_22:6","HexCer 36:1;O2|HexCer 18:1;O2/18:0","Adenosine 5'-monophosphate","SM 31:1;O2|SM 17:1;O2/14:0","TG O-54:1|TG O-18:0_16:0_20:1","DG 33:0|DG 16:0_17:0","TG 68:7|TG 20:1_26:1_22:5","TG 49:2|TG 15:0_16:1_18:1","CE 19:1","LPC 25:0/0:0","PC 34:1|PC 16:0_18:1","CE 19:3","PE 44:1|PE 26:0_18:1","TG 60:5|TG 16:1_18:1_26:3","PC 33:2","TG O-52:6|TG O-16:0_14:0_22:6","PC O-36:1|PC O-18:0_18:1","Cer 45:1;O2|Cer 18:1;O2/27:0","TG 56:5|TG 18:0_18:1_20:4","PC 41:6|PC 19:1_22:5","TG 61:5|TG 17:0_18:1_26:4","Cer 46:1;O2|Cer 20:1;O2/26:0","TG 62:5|TG 18:1_24:1_20:3","PE P-36:0|PE P-16:0_20:0","PE P-39:2|PE P-18:0_21:2","PC O-42:4|PC O-22:0_20:4","Hex2Cer 40:1;O2|Hex2Cer 18:1;O2/22:0","SM 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16:0"],"additional_accession":[]},"is_claimable":false,"name":"Lipidomic analysis of Toxoplasma gondii extracellular vesicles across host cell types","description":"Introduction. Toxoplasma gondii is an obligate intracellular parasite with an exceptional capacity to colonize a broad range of host species and cell types. Successful infection depends on its ability to manipulate host metabolism, including lipid pathways that are essential for membrane biogenesis, signalling, and immune modulation. Extracellular vesicles (EVs) are increasingly recognized as critical mediators of parasite–host interactions, but while their protein and nucleic acid cargo has been studied, the lipid composition of T. gondii EVs (TgEVs) remains poorly defined.
Methods. In this study, we performed a comprehensive lipidomic analysis of TgEVs secreted by tachyzoites grown in four distinct host cell environments: fibroblasts, Vero cells, myoblasts, and porcine intestinal epithelial cells (IPEC). Cells and TgEVs were isolated from five biological replicates per condition and analysed by liquid chromatography coupled to high-resolution tandem mass spectrometry. Comparative lipid profiling of TgEVs and their corresponding host cells was performed after total ion current normalization, followed by principal component analysis to capture global compositional patterns and pairwise differential abundance testing to identify significantly enriched or depleted lipid species.
Results: We identified 194 lipid species across 15 classes. Despite pronounced metabolic differences among host cell types, TgEVs displayed a highly conserved and distinctive lipid profile. Glycerophospholipids such as phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were the most abundant components, while sphingolipids, including sphingomyelin and ceramides, were consistently present and likely contribute to vesicle biogenesis and cargo organization. Notably, triacylglycerols (TG) were significantly enriched in TgEVs across all host conditions, suggesting active selection of neutral lipids during vesicle formation. Correlation analyses (r = 0.21–0.34) confirmed that TgEV lipidomes diverge from their cellular origin, indicating a process of active, selective sorting rather than passive acquisition from the host membrane.
Discussion: These findings demonstrate that T. gondii actively remodels host-derived lipids into vesicles with conserved and functionally specialized compositions. This selective lipid core likely underpins key aspects of parasite–host communication, including immune modulation, nutrient acquisition, and vesicle–cell interactions. Our work fundamentally advances the molecular understanding of TgEVs and establish a foundation for future studies into how lipid-mediated signalling contributes to the complex dynamics of T. gondii infection.
","dates":{"publication":"2026-02-11","submission":"2025-11-03"},"accession":"MTBLS13253","cross_references":{}}