Publication details

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

Authors

VIDOVÁ Veronika BENEŠOVÁ Eliška KLÁNOVÁ Jana THON Vojtěch SPÁČIL Zdeněk

Year of publication 2021
Type Article in Periodical
Magazine / Source Nature Scientific Reports
MU Faculty or unit

Faculty of Science

Citation
Web https://www.nature.com/articles/s41598-021-89384-0
Doi http://dx.doi.org/10.1038/s41598-021-89384-0
Keywords EOSINOPHIL-DERIVED NEUROTOXIN; INTESTINAL INFLAMMATION; FOOD ALLERGY; CESAREAN-SECTION; GRANULE PROTEINS; BOWEL DISEASES; EARLY-LIFE; MECONIUM; GUT; ALPHA-1-ANTITRYPSIN
Description 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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