BRIEF RESEARCH REPORT article
Front. Nutr.
Sec. Nutritional Immunology
Volume 12 - 2025 | doi: 10.3389/fnut.2025.1596951
A sensitive bioassay to measure NOD1/2 activation in human serum reveals differential postprandial NOD2 activation
Provisionally accepted
- 1Department of Immunology, Institute of Nutritional Medicine, University of Hohenheim, Stuttgart, Germany
- 2Institut für Humanernährung und Lebensmittelkunde, University of Kiel, Kiel, Schleswig-Holstein, Germany
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Food intake is associated with the occurrence of components and metabolites from the gut microbiota in the bloodstream. Using a widely utilised cell-based assay to measure bacterial peptidoglycan via pattern-recognition receptor activation, we found that the performance of this assay is significantly influenced by the presence of other serum components. To address this challenge, an alternative luciferase-based reporter assay protocol was established to accurately measure NOD1 and NOD2 activation by serum samples with high sensitivity. Utilising postprandial human serum samples, we tested this assay and showed that the concentration of NOD2-activating ligands differed in the postprandial phase.Together, we provide a protocol to measure NOD1/2 activation by human serum samples and highlight a role for NOD2 in the postprandial response.
Keywords: Reporter gene assay, NLR, Peptidoglycan, food intake, immune response, innate immunity
Received: 11 Apr 2025; Accepted: 06 Jun 2025.
Copyright: © 2025 Biber, Schart, Bosy-Westphal and Kufer. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Thomas A. Kufer, Department of Immunology, Institute of Nutritional Medicine, University of Hohenheim, Stuttgart, 50935, Germany
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