Analytical and functional approaches to assess the immunogenicity of gluten proteins

Gianfranco Mamone; Luigia Di Stasio; Serena Vitale; Stefania Picascia; Carmen Gianfrani

doi:10.3389/fnut.2022.1049623

Analytical and functional approaches to assess the immunogenicity of gluten proteins

Front Nutr. 2023 Jan 18:9:1049623. doi: 10.3389/fnut.2022.1049623. eCollection 2022.

Authors

Gianfranco Mamone¹, Luigia Di Stasio¹, Serena Vitale², Stefania Picascia², Carmen Gianfrani²

Affiliations

¹ Institute of Food Science, Department of Biology, Agriculture and Food Sciences, National Research Council of Italy, Avellino, Italy.
² Institute of Biochemistry and Cell Biology, Department of Biomedical Sciences, National Research Council of Italy, Naples, Italy.

Abstract

Gluten proteins are the causative agents of celiac disease (CD), a lifelong and worldwide spread food intolerance, characterized by an autoimmune enteropathy. Gluten is a complex mixture of high homologous water-insoluble proteins, characterized by a high content of glutamine and proline amino acids that confers a marked resistance to degradation by gastrointestinal proteases. As a consequence of that, large peptides are released in the gut lumen with the potential to activate inflammatory T cells, in CD predisposed individuals. To date, several strategies aimed to detoxify gluten proteins or to develop immunomodulatory drugs to recover immune tolerance to gluten are under investigation. This review overviews the state of art of both analytical and functional methods currently used to assess the immunogenicity potential of gluten proteins from different cereal sources, including native raw seed flours and complex food products, as well as drug-treated samples. The analytical design to assess the content and profile of gluten immunogenic peptides, described herein, is based on the oral-gastro-intestinal digestion (INFOGEST model) followed by extensive characterization of residual gluten peptides by proteomic and immunochemical analyses. These approaches include liquid chromatography-high-resolution mass spectrometry (LC-MS/MS) and R5/G12 competitive ELISA. Functional studies to assess the immune stimulatory capabilities of digested gluten peptides are based on gut mucosa T cells or peripheral blood cells obtained from CD volunteers after a short oral gluten challenge.

Keywords: INFOGEST digestion method; T-cell assays; celiac disease; gastrointestinal proteases; gluten immunogenic peptides (GIP); short gluten challenge.

Publication types

Review

Grants and funding

This project was received funding from the European Union’s Horizon 2020 Research and Innovation Program under the ERA-NET Cofund action N° 696296 by the Joint Programming Initiative “A Healthy Diet for a Healthy Life” (JPI HDHL) and the Federal Ministry of Education and Research Germany (BMBF), the Research Council of Norway (RCN) and the Ministry of Agricultural, Food and Forestry Policies Italy (MIPAAF -D.M. N. 623748), and the Fondazione Italiana Celiachia (Research grant N. 002_FC_2020 to CG and research fellowship N. 009_FC_2016 to SV).