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

Abstract 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....

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Autores principales:	Veronika Vidova, Eliska Benesova, Jana Klanova, Vojtech Thon, Zdenek Spacil
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8f20ac21b67445f69d239d7e5d8c7229
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spelling	oai:doaj.org-article:8f20ac21b67445f69d239d7e5d8c72292021-12-02T15:43:23ZSimultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation10.1038/s41598-021-89384-02045-2322https://doaj.org/article/8f20ac21b67445f69d239d7e5d8c72292021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-89384-0https://doaj.org/toc/2045-2322Abstract 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 µg/mg of total protein), coefficient of determination (R2; > 0.99), the limit of detection (LOD; 0.003 to 0.04 µg/mg of total protein), the limit of quantification (LOQ; 0.009 to 0.122 µg/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.Veronika VidovaEliska BenesovaJana KlanovaVojtech ThonZdenek SpacilNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Veronika Vidova Eliska Benesova Jana Klanova Vojtech Thon Zdenek Spacil Simultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation
description	Abstract 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 µg/mg of total protein), coefficient of determination (R2; > 0.99), the limit of detection (LOD; 0.003 to 0.04 µg/mg of total protein), the limit of quantification (LOQ; 0.009 to 0.122 µg/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.
format	article
author	Veronika Vidova Eliska Benesova Jana Klanova Vojtech Thon Zdenek Spacil
author_facet	Veronika Vidova Eliska Benesova Jana Klanova Vojtech Thon Zdenek Spacil
author_sort	Veronika Vidova
title	Simultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation
title_short	Simultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation
title_full	Simultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation
title_fullStr	Simultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation
title_full_unstemmed	Simultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation
title_sort	simultaneous quantitative profiling of clinically relevant immune markers in neonatal stool swabs to reveal inflammation
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8f20ac21b67445f69d239d7e5d8c7229
work_keys_str_mv	AT veronikavidova simultaneousquantitativeprofilingofclinicallyrelevantimmunemarkersinneonatalstoolswabstorevealinflammation AT eliskabenesova simultaneousquantitativeprofilingofclinicallyrelevantimmunemarkersinneonatalstoolswabstorevealinflammation AT janaklanova simultaneousquantitativeprofilingofclinicallyrelevantimmunemarkersinneonatalstoolswabstorevealinflammation AT vojtechthon simultaneousquantitativeprofilingofclinicallyrelevantimmunemarkersinneonatalstoolswabstorevealinflammation AT zdenekspacil simultaneousquantitativeprofilingofclinicallyrelevantimmunemarkersinneonatalstoolswabstorevealinflammation
_version_	1718385820993323008

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

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