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  • Coeliac disease: immunogenicity studies of barley hordein and rye secalin-derived peptides.

Coeliac disease: immunogenicity studies of barley hordein and rye secalin-derived peptides.

International journal of experimental pathology (2016-09-24)
Widya A Wahab, Tanja Šuligoj, Julia Ellis, Beatriz Côrtez-Real, Paul J Ciclitira
ABSTRACT

Coeliac disease (CD) is an inflammatory disorder of the small intestine. It includes aberrant adaptive immunity with presentation of CD toxic gluten peptides by HLA-DQ2 or DQ8 molecules to gluten-sensitive T cells. A ω-gliadin/C-hordein peptide (QPFPQPEQPFPW) and a rye-derived secalin peptide (QPFPQPQQPIPQ) were proposed to be toxic in CD, as they yielded positive responses when assessed with peripheral blood T-cell clones derived from individuals with CD. We sought to assess the immunogenicity of the candidate peptides using gluten-sensitive T-cell lines obtained from CD small intestinal biopsies. We also sought to investigate the potential cross-reactivity of wheat gluten-sensitive T-cell lines with peptic-tryptic digested barley hordein (PTH) and rye secalin (PTS). Synthesised candidate peptides were deamidated with tissue transglutaminase (tTG). Gluten-sensitive T-cell lines were generated by culturing small intestinal biopsies from CD patients with peptic-tryptic gluten (PTG), PTH or PTS, along with autologous PBMCs for antigen presentation. The stimulation indices were determined by measuring the relative cellular proliferation via incorporation of

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Trypsin Agarose, buffered aqueous suspension, from bovine pancreas trypsin
Sigma-Aldrich
Transglutaminase from guinea pig liver, lyophilized powder, ≥1.5 units/mg protein
Sigma-Aldrich
Pepsin−Agarose from porcine gastric mucosa, lyophilized powder, ≥30 units/mg dry solid
Sigma-Aldrich
HEPES solution, 1 M, pH 7.0-7.6, sterile-filtered, BioReagent, suitable for cell culture