#METABOLOMICS WORKBENCH EllenDePaepe_20230706_002000 DATATRACK_ID:4143 STUDY_ID:ST002944 ANALYSIS_ID:AN004829 PROJECT_ID:PR001831 VERSION 1 CREATED_ON October 27, 2023, 10:00 am #PROJECT PR:PROJECT_TITLE Integrated gut metabolome and microbiome fingerprinting reveals that dysbiosis PR:PROJECT_TITLE precedes allergic inflammation in IgE-mediated pediatric cow’s milk allergy PR:PROJECT_TYPE (un)targeted MS PR:PROJECT_SUMMARY Background: IgE-mediated cow’s milk allergy (IgE-CMA) is one of the first PR:PROJECT_SUMMARY allergies to arise in early childhood and may result from exposure to various PR:PROJECT_SUMMARY milk allergens, of which β-lactoglobulin (BLG) and casein are the most PR:PROJECT_SUMMARY important. Understanding the underlying mechanisms behind IgE-CMA is imperative PR:PROJECT_SUMMARY for the discovery of novel biomarkers and the design of innovative treatment and PR:PROJECT_SUMMARY prevention strategies. Methods: We report a longitudinal in vivo murine model, PR:PROJECT_SUMMARY in which 2 mice strains (BALB/c and C57Bl/6) were sensitized to BLG using either PR:PROJECT_SUMMARY cholera toxin or an oil emulsion (n=6 per group). After sensitization, mice were PR:PROJECT_SUMMARY challenged orally, their clinical signs monitored, antibody (IgE and IgG1) and PR:PROJECT_SUMMARY cytokine levels (IL-4 and IFN-γ) measured, and fecal samples subjected to PR:PROJECT_SUMMARY metabolomics. The results of the murine models were further supported by fecal PR:PROJECT_SUMMARY microbiome-metabolome data from our population of IgE-CMA (n=24) and healthy PR:PROJECT_SUMMARY (n=23) children (Trial: NCT04249973), on which polar metabolomics, lipidomics PR:PROJECT_SUMMARY and 16S rRNA metasequencing were performed. In vitro gastrointestinal digestions PR:PROJECT_SUMMARY and multi-omics corroborated the microbial origin of proposed metabolic changes. PR:PROJECT_SUMMARY Results: During sensitization, we observed multiple microbially derived PR:PROJECT_SUMMARY metabolic alterations, most importantly bile acid, energy and tryptophan PR:PROJECT_SUMMARY metabolites, that preceded allergic inflammation. The latter was reflected in a PR:PROJECT_SUMMARY disturbed sphingolipid metabolism. We confirmed microbial dysbiosis, and its PR:PROJECT_SUMMARY causal effect on metabolic alterations in our patient cohort, which was PR:PROJECT_SUMMARY accompanied by metabolic signatures of low-grade inflammation. Conclusion: Our PR:PROJECT_SUMMARY results indicate that gut dysbiosis precedes allergic inflammation and nurtures PR:PROJECT_SUMMARY a chronic low-grade inflammation in children on elimination diets, opening PR:PROJECT_SUMMARY important new opportunities for future prevention and treatment strategies. PR:INSTITUTE Ghent University PR:DEPARTMENT Translational Physiology, Infectiology and Public Health PR:LABORATORY Laboratory for Integrative Metabolomics PR:LAST_NAME De Paepe PR:FIRST_NAME Ellen PR:ADDRESS Salisburylaan 133, Merelbeke, Oost-Vlaanderen, 9820, Belgium PR:EMAIL Ellen.DePaepe@UGent.be PR:PHONE 0032479081098 PR:FUNDING_SOURCE Fonds Wetenschappelijk Onderzoek #STUDY ST:STUDY_TITLE Longitudinal polar fecal metabolomics of mice undergoing sensitization to ST:STUDY_TITLE beta-lactoglobulin ST:STUDY_SUMMARY This study is part of a multi-part study, including a. longitudinal polar fecal ST:STUDY_SUMMARY metabolomics of mice undergoing sensitization to beta-lactoglobulin b. polar ST:STUDY_SUMMARY fecal metabolomics of a patient cohort c. fecal lipidomics of a patient cohort ST:STUDY_SUMMARY d. polar urinary metabolomics of a patient cohort e. polar metabolomics of in ST:STUDY_SUMMARY vitro digestions This specific part is part a. longitudinal polar fecal ST:STUDY_SUMMARY metabolomics of mice undergoing sensitization to beta-lactoglobulin ST:INSTITUTE Ghent University ST:DEPARTMENT Department of Translational Physiology, Infectiology and Public Health ST:LABORATORY Laboratory for Integrative Metabolomics ST:LAST_NAME De Paepe ST:FIRST_NAME Ellen ST:ADDRESS Salisburylaan 133, 9820 Merelbeke, Belgium ST:EMAIL Ellen.DePaepe@UGent.be ST:STUDY_TYPE (un)targeted MS ST:PHONE 0032479081098 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN BALB/c and C57Bl/6 SU:AGE_OR_AGE_RANGE 3-10 weeks SU:GENDER Female SU:ANIMAL_ANIMAL_SUPPLIER Charles River SU:ANIMAL_HOUSING SPF SU:ANIMAL_FEED Sniff SU:ANIMAL_WATER Ad libitum #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 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iQC21 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se163.raw SUBJECT_SAMPLE_FACTORS - iQC22 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se164.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_126 Strain:B6 | treatment:2W | week:Wk 1 RAW_FILE_NAME=210611_se166.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_128 Strain:B6 | treatment:6D | week:Wk 1 RAW_FILE_NAME=210611_se167.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_129 Strain:B6 | treatment:1C | week:Wk 1 RAW_FILE_NAME=210611_se168.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_130 Strain:BC | treatment:4W | week:Wk 3 RAW_FILE_NAME=210611_se169.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_131 Strain:BC | treatment:4E | week:Wk 3 RAW_FILE_NAME=210611_se170.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_132 Strain:B6 | treatment:3A | week:Wk 4 RAW_FILE_NAME=210611_se171.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_134 Strain:BC | treatment:3W | week:Wk 3 RAW_FILE_NAME=210611_se172.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_136 Strain:BC | treatment:6E | week:Wk 2 RAW_FILE_NAME=210611_se173.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_137 Strain:BC | treatment:5Z | week:Wk 5 RAW_FILE_NAME=210611_se174.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_138 Strain:BC | treatment:3B | week:Wk 2 RAW_FILE_NAME=210611_se175.raw SUBJECT_SAMPLE_FACTORS - iQC23 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se176.raw SUBJECT_SAMPLE_FACTORS - iQC24 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se177.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_139 Strain:B6 | treatment:5Z | week:Wk 7 RAW_FILE_NAME=210611_se179.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_140 Strain:BC | treatment:2E | week:Wk 1 RAW_FILE_NAME=210611_se180.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_141 Strain:BC | treatment:4E | week:Wk 1 RAW_FILE_NAME=210611_se181.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_142 Strain:BC | treatment:5B | week:Wk 4 RAW_FILE_NAME=210611_se182.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_143 Strain:BC | treatment:3A | week:Wk 3 RAW_FILE_NAME=210611_se183.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_144 Strain:BC | treatment:2Z | week:Wk 7 RAW_FILE_NAME=210611_se184.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_145 Strain:BC | treatment:2A | week:Wk 2 RAW_FILE_NAME=210611_se185.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_146 Strain:BC | treatment:3X | week:Wk 7 RAW_FILE_NAME=210611_se186.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_147 Strain:BC | treatment:6E | week:Wk 1 RAW_FILE_NAME=210611_se187.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_149 Strain:BC | treatment:6Z | week:Wk 1 RAW_FILE_NAME=210611_se188.raw SUBJECT_SAMPLE_FACTORS - iQC25 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se189.raw SUBJECT_SAMPLE_FACTORS - iQC26 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se190.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_150 Strain:BC | treatment:5Y | week:Wk 1 RAW_FILE_NAME=210611_se192.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_151 Strain:B6 | treatment:4A | week:Wk 4 RAW_FILE_NAME=210611_se193.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_152 Strain:BC | treatment:4A | week:Wk 3 RAW_FILE_NAME=210611_se194.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_153 Strain:BC | treatment:4A | week:Wk 2 RAW_FILE_NAME=210611_se195.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_155 Strain:BC | treatment:3C | week:Wk 1 RAW_FILE_NAME=210611_se196.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_156 Strain:BC | treatment:5Z | week:Wk 7 RAW_FILE_NAME=210611_se197.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_157 Strain:BC | treatment:6F | week:Wk 3 RAW_FILE_NAME=210611_se198.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_158 Strain:B6 | treatment:2E | week:Wk 4 RAW_FILE_NAME=210611_se199.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_159 Strain:BC | treatment:2A | week:Wk 3 RAW_FILE_NAME=210611_se200.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_160 Strain:BC | treatment:3Z | week:Wk 5 RAW_FILE_NAME=210611_se201.raw SUBJECT_SAMPLE_FACTORS - iQC27 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se202.raw SUBJECT_SAMPLE_FACTORS - iQC28 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se203.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_161 Strain:BC | treatment:4B | week:Wk 2 RAW_FILE_NAME=210611_se207.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_162 Strain:BC | treatment:6B | week:Wk 2 RAW_FILE_NAME=210611_se208.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_163 Strain:BC | treatment:6W | week:Wk 7 RAW_FILE_NAME=210611_se209.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_164 Strain:BC | treatment:2E | week:Wk 3 RAW_FILE_NAME=210611_se210.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_166 Strain:BC | treatment:1W | week:Wk 5 RAW_FILE_NAME=210611_se211.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_167 Strain:BC | treatment:1E | week:Wk 2 RAW_FILE_NAME=210611_se212.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_168 Strain:B6 | treatment:4Y | week:Wk 1 RAW_FILE_NAME=210611_se213.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_169 Strain:BC | treatment:3F | week:Wk 4 RAW_FILE_NAME=210611_se214.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_170 Strain:BC | treatment:6Z | week:Wk 3 RAW_FILE_NAME=210611_se215.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_171 Strain:BC | treatment:6E | week:Wk 4 RAW_FILE_NAME=210611_se216.raw SUBJECT_SAMPLE_FACTORS - iQC29 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se217.raw SUBJECT_SAMPLE_FACTORS - iQC30 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se218.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_172 Strain:B6 | treatment:4D | week:Wk 4 RAW_FILE_NAME=210611_se220.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_173 Strain:BC | treatment:6A | week:Wk 2 RAW_FILE_NAME=210611_se221.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_174 Strain:B6 | treatment:5X | week:Wk 7 RAW_FILE_NAME=210611_se222.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_175 Strain:BC | treatment:5C | week:Wk 4 RAW_FILE_NAME=210611_se223.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_176 Strain:BC | treatment:6W | week:Wk 3 RAW_FILE_NAME=210611_se224.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_177 Strain:BC | treatment:2W | week:Wk 1 RAW_FILE_NAME=210611_se225.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_178 Strain:BC | treatment:1Y | week:Wk 7 RAW_FILE_NAME=210611_se226.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_179 Strain:B6 | treatment:1B | week:Wk 4 RAW_FILE_NAME=210611_se227.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_180 Strain:BC | treatment:6C | week:Wk 4 RAW_FILE_NAME=210611_se228.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_181 Strain:BC | treatment:5A | week:Wk 1 RAW_FILE_NAME=210611_se229.raw SUBJECT_SAMPLE_FACTORS - iQC31 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se230.raw SUBJECT_SAMPLE_FACTORS - iQC32 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se231.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_184 Strain:B6 | treatment:5B | week:Wk 4 RAW_FILE_NAME=210611_se233.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_185 Strain:BC | treatment:2X | week:Wk 7 RAW_FILE_NAME=210611_se234.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_187 Strain:BC | treatment:1F | week:Wk 4 RAW_FILE_NAME=210611_se235.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_188 Strain:BC | treatment:6X | week:Wk 7 RAW_FILE_NAME=210611_se236.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_189 Strain:BC | treatment:5D | week:Wk 1 RAW_FILE_NAME=210611_se237.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_190 Strain:B6 | treatment:1X | week:Wk 1 RAW_FILE_NAME=210611_se238.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_191 Strain:B6 | treatment:1A | week:Wk 4 RAW_FILE_NAME=210611_se239.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_192 Strain:BC | treatment:3D | week:Wk 1 RAW_FILE_NAME=210611_se240.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_193 Strain:B6 | treatment:2C | week:Wk 4 RAW_FILE_NAME=210611_se241.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_194 Strain:BC | treatment:2Z | week:Wk 1 RAW_FILE_NAME=210611_se242.raw SUBJECT_SAMPLE_FACTORS - iQC33 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se243.raw SUBJECT_SAMPLE_FACTORS - iQC34 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se244.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_195 Strain:B6 | treatment:2B | week:Wk 4 RAW_FILE_NAME=210611_se246.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_197 Strain:BC | treatment:3C | week:Wk 4 RAW_FILE_NAME=210611_se247.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_198 Strain:B6 | treatment:4D | week:Wk 1 RAW_FILE_NAME=210611_se248.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_199 Strain:B6 | treatment:6W | week:Wk 7 RAW_FILE_NAME=210611_se249.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_200 Strain:BC | treatment:6D | week:Wk 1 RAW_FILE_NAME=210611_se250.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_201 Strain:BC | treatment:1B | week:Wk 3 RAW_FILE_NAME=210611_se251.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_202 Strain:B6 | treatment:4C | week:Wk 1 RAW_FILE_NAME=210611_se252.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_204 Strain:BC | treatment:1W | week:Wk 7 RAW_FILE_NAME=210611_se253.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_205 Strain:B6 | treatment:1C | week:Wk 4 RAW_FILE_NAME=210611_se254.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_206 Strain:B6 | treatment:5F | week:Wk 4 RAW_FILE_NAME=210611_se255.raw SUBJECT_SAMPLE_FACTORS - iQC35 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se256.raw SUBJECT_SAMPLE_FACTORS - iQC36 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se257.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_207 Strain:B6 | treatment:2Y | week:Wk 7 RAW_FILE_NAME=210611_se261.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_208 Strain:BC | treatment:1D | week:Wk 4 RAW_FILE_NAME=210611_se262.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_209 Strain:B6 | treatment:5E | week:Wk 4 RAW_FILE_NAME=210611_se263.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_210 Strain:BC | treatment:1A | week:Wk 3 RAW_FILE_NAME=210611_se264.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_211 Strain:B6 | treatment:2C | week:Wk 1 RAW_FILE_NAME=210611_se265.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_212 Strain:BC | treatment:3F | week:Wk 2 RAW_FILE_NAME=210611_se266.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_213 Strain:BC | treatment:6F | week:Wk 1 RAW_FILE_NAME=210611_se267.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_214 Strain:B6 | treatment:6F | week:Wk 4 RAW_FILE_NAME=210611_se268.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_215 Strain:B6 | treatment:6Y | week:Wk 1 RAW_FILE_NAME=210611_se269.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_216 Strain:BC | treatment:6B | week:Wk 3 RAW_FILE_NAME=210611_se270.raw SUBJECT_SAMPLE_FACTORS - iQC37 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se271.raw SUBJECT_SAMPLE_FACTORS - iQC38 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se272.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_217 Strain:BC | treatment:2F | week:Wk 2 RAW_FILE_NAME=210611_se274.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_218 Strain:BC | treatment:5F | week:Wk 4 RAW_FILE_NAME=210611_se275.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_219 Strain:BC | treatment:3Y | week:Wk 1 RAW_FILE_NAME=210611_se276.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_220 Strain:BC | treatment:1F | week:Wk 2 RAW_FILE_NAME=210611_se277.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_221 Strain:BC | treatment:3W | week:Wk 5 RAW_FILE_NAME=210611_se278.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_222 Strain:B6 | treatment:3C | week:Wk 4 RAW_FILE_NAME=210611_se279.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_223 Strain:BC | treatment:6Y | week:Wk 1 RAW_FILE_NAME=210611_se280.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_224 Strain:B6 | treatment:6Z | week:Wk 7 RAW_FILE_NAME=210611_se281.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_225 Strain:BC | treatment:6B | week:Wk 1 RAW_FILE_NAME=210611_se282.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_226 Strain:BC | treatment:1B | week:Wk 2 RAW_FILE_NAME=210611_se283.raw SUBJECT_SAMPLE_FACTORS - iQC39 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se284.raw SUBJECT_SAMPLE_FACTORS - iQC40 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se285.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_227 Strain:BC | treatment:1A | week:Wk 4 RAW_FILE_NAME=210611_se287.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_229 Strain:B6 | treatment:4F | week:Wk 4 RAW_FILE_NAME=210611_se288.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_230 Strain:BC | treatment:2Z | week:Wk 3 RAW_FILE_NAME=210611_se289.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_232 Strain:BC | treatment:6E | week:Wk 3 RAW_FILE_NAME=210611_se290.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_233 Strain:B6 | treatment:5A | week:Wk 4 RAW_FILE_NAME=210611_se291.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_235 Strain:BC | treatment:3D | week:Wk 4 RAW_FILE_NAME=210611_se292.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_236 Strain:B6 | treatment:1Z | week:Wk 7 RAW_FILE_NAME=210611_se293.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_237 Strain:BC | treatment:5E | week:Wk 3 RAW_FILE_NAME=210611_se294.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_238 Strain:BC | treatment:6D | week:Wk 4 RAW_FILE_NAME=210611_se295.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_239 Strain:BC | treatment:3Y | week:Wk 7 RAW_FILE_NAME=210611_se296.raw SUBJECT_SAMPLE_FACTORS - iQC41 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se297.raw SUBJECT_SAMPLE_FACTORS - iQC42 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se298.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_240 Strain:BC | treatment:2F | week:Wk 1 RAW_FILE_NAME=210611_se302.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_242 Strain:BC | treatment:4E | week:Wk 4 RAW_FILE_NAME=210611_se303.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_243 Strain:BC | treatment:4Z | week:Wk 1 RAW_FILE_NAME=210611_se304.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_244 Strain:B6 | treatment:2X | week:Wk 7 RAW_FILE_NAME=210611_se305.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_245 Strain:B6 | treatment:1Y | week:Wk 7 RAW_FILE_NAME=210611_se306.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_246 Strain:BC | treatment:2Y | week:Wk 1 RAW_FILE_NAME=210611_se307.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_251 Strain:BC | treatment:1B | week:Wk 1 RAW_FILE_NAME=210611_se308.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_252 Strain:B6 | treatment:5A | week:Wk 1 RAW_FILE_NAME=210611_se309.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_253 Strain:BC | treatment:5Z | week:Wk 3 RAW_FILE_NAME=210611_se310.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_254 Strain:B6 | treatment:3A | week:Wk 1 RAW_FILE_NAME=210611_se311.raw SUBJECT_SAMPLE_FACTORS - iQC43 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se312.raw SUBJECT_SAMPLE_FACTORS - iQC44 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se313.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_255 Strain:BC | treatment:5A | week:Wk 2 RAW_FILE_NAME=210611_se315.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_256 Strain:BC | treatment:2C | week:Wk 4 RAW_FILE_NAME=210611_se316.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_257 Strain:BC | treatment:1Z | week:Wk 1 RAW_FILE_NAME=210611_se317.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_258 Strain:BC | treatment:6Y | week:Wk 7 RAW_FILE_NAME=210611_se318.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_259 Strain:BC | treatment:4F | week:Wk 1 RAW_FILE_NAME=210611_se319.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_260 Strain:B6 | treatment:2Z | week:Wk 1 RAW_FILE_NAME=210611_se320.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_261 Strain:B6 | treatment:4X | week:Wk 1 RAW_FILE_NAME=210611_se321.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_262 Strain:B6 | treatment:3X | week:Wk 7 RAW_FILE_NAME=210611_se322.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_263 Strain:B6 | treatment:2X | week:Wk 1 RAW_FILE_NAME=210611_se323.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_264 Strain:B6 | treatment:3E | week:Wk 4 RAW_FILE_NAME=210611_se324.raw SUBJECT_SAMPLE_FACTORS - iQC45 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se325.raw SUBJECT_SAMPLE_FACTORS - iQC46 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se326.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_265 Strain:BC | treatment:3E | week:Wk 2 RAW_FILE_NAME=210611_se328.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_266 Strain:B6 | treatment:6C | week:Wk 4 RAW_FILE_NAME=210611_se329.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_267 Strain:B6 | treatment:2A | week:Wk 1 RAW_FILE_NAME=210611_se330.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_268 Strain:B6 | treatment:4E | week:Wk 1 RAW_FILE_NAME=210611_se331.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_269 Strain:BC | treatment:1A | week:Wk 2 RAW_FILE_NAME=210611_se332.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_270 Strain:BC | treatment:6A | week:Wk 4 RAW_FILE_NAME=210611_se333.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_271 Strain:B6 | treatment:3X | week:Wk 1 RAW_FILE_NAME=210611_se334.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_273 Strain:B6 | treatment:1X | week:Wk 7 RAW_FILE_NAME=210611_se335.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_274 Strain:B6 | treatment:5W | week:Wk 1 RAW_FILE_NAME=210611_se336.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_275 Strain:BC | treatment:5A | week:Wk 4 RAW_FILE_NAME=210611_se337.raw SUBJECT_SAMPLE_FACTORS - iQC47 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se338.raw SUBJECT_SAMPLE_FACTORS - iQC48 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se339.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_276 Strain:BC | treatment:4A | week:Wk 1 RAW_FILE_NAME=210611_se341.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_277 Strain:BC | treatment:2D | week:Wk 1 RAW_FILE_NAME=210611_se342.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_278 Strain:BC | treatment:6A | week:Wk 1 RAW_FILE_NAME=210611_se343.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_279 Strain:B6 | treatment:6E | week:Wk 1 RAW_FILE_NAME=210611_se344.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_280 Strain:BC | treatment:1C | week:Wk 4 RAW_FILE_NAME=210611_se345.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_281 Strain:BC | treatment:2W | week:Wk 5 RAW_FILE_NAME=210611_se346.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_282 Strain:B6 | treatment:3F | week:Wk 4 RAW_FILE_NAME=210611_se347.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_283 Strain:BC | treatment:1Z | week:Wk 5 RAW_FILE_NAME=210611_se348.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_285 Strain:BC | treatment:3A | week:Wk 4 RAW_FILE_NAME=210611_se349.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_286 Strain:B6 | treatment:4W | week:Wk 7 RAW_FILE_NAME=210611_se350.raw SUBJECT_SAMPLE_FACTORS - iQC49 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se351.raw SUBJECT_SAMPLE_FACTORS - iQC50 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se352.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_287 Strain:B6 | treatment:5Z | week:Wk 1 RAW_FILE_NAME=210611_se370.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_288 Strain:B6 | treatment:4Z | week:Wk 7 RAW_FILE_NAME=210611_se371.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_289 Strain:BC | treatment:3F | week:Wk 1 RAW_FILE_NAME=210611_se372.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_290 Strain:BC | treatment:5B | week:Wk 3 RAW_FILE_NAME=210611_se373.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_291 Strain:BC | treatment:1X | week:Wk 7 RAW_FILE_NAME=210611_se374.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_293 Strain:BC | treatment:1A | week:Wk 1 RAW_FILE_NAME=210611_se375.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_294 Strain:B6 | treatment:3W | week:Wk 7 RAW_FILE_NAME=210611_se376.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_295 Strain:BC | treatment:5W | week:Wk 5 RAW_FILE_NAME=210611_se377.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_296 Strain:BC | treatment:4B | week:Wk 1 RAW_FILE_NAME=210611_se378.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_297 Strain:BC | treatment:4Y | week:Wk 1 RAW_FILE_NAME=210611_se379.raw SUBJECT_SAMPLE_FACTORS - iQC53 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se380.raw SUBJECT_SAMPLE_FACTORS - iQC54 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se381.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_298 Strain:BC | treatment:4C | week:Wk 1 RAW_FILE_NAME=210611_se383.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_299 Strain:BC | treatment:5Y | week:Wk 7 RAW_FILE_NAME=210611_se384.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_300 Strain:BC | treatment:3A | week:Wk 2 RAW_FILE_NAME=210611_se385.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_301 Strain:BC | treatment:5D | week:Wk 4 RAW_FILE_NAME=210611_se386.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_302 Strain:BC | treatment:1W | week:Wk 3 RAW_FILE_NAME=210611_se387.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_303 Strain:BC | treatment:4F | week:Wk 3 RAW_FILE_NAME=210611_se388.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_304 Strain:B6 | treatment:6Z | week:Wk 1 RAW_FILE_NAME=210611_se389.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_305 Strain:BC | treatment:4W | week:Wk 5 RAW_FILE_NAME=210611_se390.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_306 Strain:B6 | treatment:5Y | week:Wk 7 RAW_FILE_NAME=210611_se391.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_308 Strain:B6 | treatment:1Z | week:Wk 1 RAW_FILE_NAME=210611_se392.raw SUBJECT_SAMPLE_FACTORS - iQC55 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se393.raw SUBJECT_SAMPLE_FACTORS - iQC56 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se394.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_309 Strain:BC | treatment:4B | week:Wk 3 RAW_FILE_NAME=210611_se396.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_310 Strain:BC | treatment:3W | week:Wk 1 RAW_FILE_NAME=210611_se397.raw SUBJECT_SAMPLE_FACTORS - S_21_0066_311 Strain:B6 | treatment:5W | week:Wk 7 RAW_FILE_NAME=210611_se398.raw SUBJECT_SAMPLE_FACTORS - iQC57 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se399.raw SUBJECT_SAMPLE_FACTORS - iQC58 Strain:- | treatment:- | week:- RAW_FILE_NAME=210611_se400.raw #COLLECTION CO:COLLECTION_SUMMARY Fecal samples were collected on different timepoints during sensitization of the CO:COLLECTION_SUMMARY mice. Samples were freeze-dried within one week after collection and stored at CO:COLLECTION_SUMMARY -80° awaiting analysis CO:SAMPLE_TYPE Feces CO:COLLECTION_METHOD Spontaneous defecation CO:COLLECTION_FREQUENCY 1x week CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Two different mice strains and two different protocols were tested: BALB/c (BC) TR:TREATMENT_SUMMARY mice and C57Bl/6 (B6) mice. The first sensitization protocol was based on TR:TREATMENT_SUMMARY Adel-Patient et al., in which 4-week-old mice received oral gavages containing TR:TREATMENT_SUMMARY ß-lactoglobulin (BLG) and cholera toxin (CT) once a week for 6 weeks 20 TR:TREATMENT_SUMMARY (treatment W), the control groups received either BLG in PBS (X), CT in PBS (Y) TR:TREATMENT_SUMMARY or PBS alone (Z). The second protocol was adapted from Shindo et al. and TR:TREATMENT_SUMMARY included intragastrical administration of BLG in PBS or oil emulsion, followed TR:TREATMENT_SUMMARY by intraperitoneal injection of sodium salicylate (SA). The latter protocol was TR:TREATMENT_SUMMARY performed 5x a week for 3 weeks. Group A received BLG in PBS + SA, group B BLG TR:TREATMENT_SUMMARY in oil + SA, group C BLG in PBS + PBS, group D BLG in oil + PBS, group E and F TR:TREATMENT_SUMMARY were vehicle controls with group E receiving PBS + SA, and group F oil + SA TR:TREATMENT_COMPOUND Beta-lactoglobulin, cholera toxin TR:TREATMENT_ROUTE oral gavage, intraperitoneal injection TR:TREATMENT_DOSE 1 mg ß-lactoglobulin (BLG) per g body weight mixed with 10 µg of cholera toxin TR:TREATMENT_DOSE (CT) in 0.1 mL PBS, or 1 mg BLG in 0.1 mL individual antigen solution (vehicle), TR:TREATMENT_DOSE which was either PBS or an oil emulsion (saline, linoleic acid, and lecithin at TR:TREATMENT_DOSE 5:4:1 by volume). Intraperitoneal administration of 0.1 mL PBS or PBS containing TR:TREATMENT_DOSE 0.2 mg sodium salicylate (SA) TR:TREATMENT_DOSEDURATION 3 or 7 weeks TR:TREATMENT_VEHICLE PBS or oil emulsion TR:ANIMAL_ANESTHESIA CO TR:ANIMAL_FASTING Evening before antigen challenge TR:ANIMAL_ENDP_EUTHANASIA cervical dislocation TR:ANIMAL_ENDP_CLINICAL_SIGNS severe anaphylaxis #SAMPLEPREP SP:SAMPLEPREP_SUMMARY To extract the polar fecal metabolome, 200 mg of lyophilized homogenized feces SP:SAMPLEPREP_SUMMARY was dissolved in 4 mL of ultrapure water, after the addition of 100 mL ISTD SP:SAMPLEPREP_SUMMARY mixture (25 ng mL&1 of Dvaline- d8 and L-alanine-d3). Subsequent to 30 s of SP:SAMPLEPREP_SUMMARY thorough vortexing, 1 mL of an ice-cold methanol and ultrapure water (80:20, v/ SP:SAMPLEPREP_SUMMARY v) mixture was added. The supernatant of the solid-liquid extraction was SP:SAMPLEPREP_SUMMARY collected after 1 min of vortexing and 10 min of rotation, followed by a 10-min SP:SAMPLEPREP_SUMMARY centrifugation step (13 300 x g, at 4 %C). Next, the extract was passed over a SP:SAMPLEPREP_SUMMARY polyamide filter (diameter of 25mm and pore size of 0.45 mm) (Machery-Nagel, SP:SAMPLEPREP_SUMMARY Düren, Germany), diluted (1:3) with ultrapure water and transferred to a glass SP:SAMPLEPREP_SUMMARY HPLCvial. SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:SAMPLE_SPIKING Internal standard #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) CH:SOLVENT_A 100% water; 0.1% formic acid CH:SOLVENT_B 100% acetonitrile; 0.1% formic acid CH:FLOW_GRADIENT A gradient profile with following proportions (v/v) of solvent A was applied: CH:FLOW_GRADIENT 0e1.5 min at 98%, 1.5e7.0 min from 98% to 75%, 7.0e8.0 min from 75% to 40%, CH:FLOW_GRADIENT 8.0e12.0 min from 40% to 5%, 12.0e14.0 min at 5%, 14.0e14.1 min from 5 to 98%, CH:FLOW_GRADIENT followed by 4.0 min of re-equilibration. CH:FLOW_RATE 0.4 mL/min CH:COLUMN_TEMPERATURE 45 CH:INTERNAL_STANDARD L-alanine-d3, D-valine-d8 CH:SAMPLE_INJECTION 10 uL #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS MS acquisition: Xcalibur Targeted data processing: Xcalibur Untargeted data MS:MS_COMMENTS acquisition: Compound Discoverer, R studio MS:MS_RESULTS_FILE ST002944_AN004829_Results.txt UNITS:a.u. Has m/z:Neutral masses Has RT:Yes RT units:Minutes #END