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Simultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation

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VIDOVÁ Veronika BENEŠOVÁ Eliška KLÁNOVÁ Jana THON Vojtěch SPÁČIL Zdeněk

Rok publikování 2021
Druh Článek v odborném periodiku
Časopis / Zdroj Nature Scientific Reports
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
www https://www.nature.com/articles/s41598-021-89384-0
Doi http://dx.doi.org/10.1038/s41598-021-89384-0
Klíčová slova EOSINOPHIL-DERIVED NEUROTOXIN; INTESTINAL INFLAMMATION; FOOD ALLERGY; CESAREAN-SECTION; GRANULE PROTEINS; BOWEL DISEASES; EARLY-LIFE; MECONIUM; GUT; ALPHA-1-ANTITRYPSIN
Popis An aberrant immune response developed early in life may trigger inflammatory bowel disease (IBD) and food allergies (e.g., celiac disease). Fecal levels of immune markers categorize an inflammatory response (e.g., food allergy, autoimmune) paralleled with the initial microbial colonization. The immunoaffinity assays are routinely applied to quantify circulating immune protein markers in blood/serum. However, a reliable, multiplex assay to quantify fecal levels of immune proteins is unavailable. We developed mass spectrometry assays to simultaneously quantify fecal calprotectin, myeloperoxidase, eosinophil-derived neurotoxin, eosinophil cationic protein, alpha-1-antitrypsin 1, and adaptive immunity effectors in 134 neonatal stool swabs. We optimized extraction and proteolytic protocol and validated the multiplex assay in terms of linearity of response (>100; typically 0.04 to 14.77 mu g/mg of total protein), coefficient of determination (R-2;>0.99), the limit of detection (LOD; 0.003 to 0.04 mu g/mg of total protein), the limit of quantification (LOQ; 0.009 to 0.122 mu g/mg of total protein) and robustness. The median CV of intra- and interday precision was 9.8% and 14.1%, respectively. We quantified breast milk-derived IGHA2 to differentiate meconium from feces samples and to detect the first food intake. An early life profiling of immune markers reflects disrupted intestinal homeostasis, and it is perhaps suitable for pre-symptomatic interception of IBD and food allergies.
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