The In Vitro Effects of Enzymatic Digested Gliadin on the Functionality of the Autophagy Process

• Main Authors: Federico Manai, Alberto Azzalin, Fabio Gabriele, Carolina Martinelli, Martina Morandi, Marco Biggiogera, Mauro Bozzola, Sergio Comincini
• Format: Article
• Language: English
• Published: MDPI AG 2018-02-01
• Series: International Journal of Molecular Sciences
• Subjects: celiac disease; gluten; autophagosome; Caco-2 cells
• Online Access: http://www.mdpi.com/1422-0067/19/2/635

Gliadin, the alcohol-soluble protein fraction of wheat, contains the factor toxic for celiac disease (CD), and its toxicity is not reduced by digestion with gastro-pancreatic enzymes. Importantly, it is proved that an innate immunity to gliadin plays a key role in the development of CD. The immune response induces epithelial stress and reprograms intraepithelial lymphocytes into natural killer (NK)-like cells, leading to enterocyte apoptosis and an increase in epithelium permeability. In this contribution, we have reported that in Caco-2 cells the administration of enzymatically digested gliadin (PT-gliadin) reduced significantly the expression of the autophagy-related marker LC3-II. Furthermore, electron and fluorescent microscope analysis suggested a compromised functionality of the autophagosome apparatus. The rescue of the dysregulated autophagy process, along with a reduction of PT-gliadin toxicity, was obtained with a starvation induction protocol and by 3-methyladenine administration, while rapamycin, a well-known autophagy inducer, did not produce a significant improvement in the clearance of extra- and intra-cellular fluorescent PT-gliadin amount. Altogether, our results highlighted the possible contribution of the autophagy process in the degradation and in the reduction of extra-cellular release of gliadin peptides and suggest novel molecular targets to counteract gliadin-induced toxicity in CD.