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Journal of Diabetes Research
Volume 2016, Article ID 2424306, 11 pages
Research Article

Large Gliadin Peptides Detected in the Pancreas of NOD and Healthy Mice following Oral Administration

1The Bartholin Institute, Rigshospitalet, Copenhagen N, Denmark
2Clinical Biochemistry, Immunology & Genetics, Statens Serum Institut, Copenhagen S, Denmark
3The Hevesy Laboratory, DTU Nutech, Technical University of Denmark, Roskilde, Denmark
4Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Prague 6, Czech Republic
5Enzyme Purification and Characterization, Novozymes A/S, Bagsværd, Denmark

Received 6 May 2016; Accepted 10 August 2016

Academic Editor: Marco Songini

Copyright © 2016 Susanne W. Bruun et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Fig Supp 1. Tissue distribution of 33-mer, 19-mer and tyrosine following intravenous injection in NOD and BALB/c mice, 8-12 weeks of age. 3H-labeled 33-mer or 3H-labeled 19-mer or 3H-tyrosine was administered to 20-week-old NOD mice. Blood and organs were sampled after 1, 24 and 72 h. The specific radioactivity is shown relative to blood, and data are average values of 2-4 mice and shown with SEM values. Abbreviations are as in Fig 1.Fig Supp 2. Kinetics of 3H-33-mer and 3H-19-mer in mouse blood following i.v. and p.o. administration. 3H-33-mer (A) or 3H-19-mer (B) was given i.v. to NOD mice (6-8 weeks). Tail blood was analyzed by SDS-PAGE and fluorography at the indicated time points (min). CT: control 33-mer/19-mer. In (A, right), 660 µg of unlabeled 33-mer was injected prior to the labeled peptide. (C) 3H-33-mer was administered p.o. to NOD mice (6-8 weeks). Blood was analyzed after 0.5, 1 and 20 h. All lanes were loaded with the same amount of radioactivity. CT is control 33-mer. (D) Fluorogram of in vitro incubation of blood plasma from a C57BL/6 mouse with 33-mer (28.6 ng/µl or 222.000 dpm/µl) for 0-4 h. CT is control 33-mer. (E) Plasma from mice, that had received 3H-labelled 33-mer one hour earlier by i.v. or p.o. administration, was depleted of albumin and IgG. Radioactivity counts in gel slices from lanes, loaded with plasma before (-) and after (+) extraction, are illustrated using pseudo-colors in intervals from <100 dpm (white), through 100-400 dpm, 400-1000 dpm and 1000-2000 dpm to >2000 dpm (dark gray). Data were normalized between lanes to contain identical amounts of radioactivity outside the albumin/IgG region. (F) Plasma from a mouse, that received 1.2 mCi 3H-labeled 33-mer p.o., was digested with trypsin followed by SDS-PAGE (right) and fluorography (left).

  1. Supplementary Material