Epitopes Related to Coeliac Disease

a coeliac disease and epitope technology, applied in the field of epitopes related to coeliac disease, can solve the problems of toxic gluten proteins in wheat, rye, barley and in some cases oats, and achieve the effect of high throughpu

Inactive Publication Date: 2008-12-25
BTG INT LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0011]The current study set out to develop a method that would allow mapping of all T-cell epitopes in gluten. Consumption of wheat bread (200 g daily for 3 days) or oats (100 g daily for 3 days) was used to induce gluten or avenin-specific T-cells in peripheral blood (collected 6 days after beginning the challenge). Peripheral blood mononuclear cells (PBMC) were assessed in overnight interferon gamma ELISPOT assays using a library of gluten and avenin peptides including all unique 12mer sequences included in every Genbank entry for wheat gluten and / or oat avenins. This goal was achieved by establishing an algorithm to design peptides spanning all potential epitopes in gluten proteins in Genbank (2922 20mers included all 14 964 unique 9mers—potential T-cell epitopes), adapting the interferon-gamma ELISPOT assay to a high throughput assay capable of screening over 1000 peptides with a single individual's blood and developing bioinformatics tools to analyse and interpret the data generated.

Problems solved by technology

Gluten proteins in wheat, rye, barley and in some cases oats are toxic in coeliac disease.

Method used

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  • Epitopes Related to Coeliac Disease
  • Epitopes Related to Coeliac Disease
  • Epitopes Related to Coeliac Disease

Examples

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examples

[0210]The invention is illustrated by the following nonlimiting Examples:

Initial Gliadin Epitope Screening Library

[0211]In initial experiments involving 29 HLA-DQ2+ individuals with coeliac disease on long-term gluten free diet, interferon-gamma ELISPOT assays were used to screen a previous Pepset (described in WO 03 / 104273, which is incorporated herein by reference) initially as pools of peptides and then in 15 subjects as individual peptides with and without deamidation by tTG. This Pepset library consisted of 652 20mer gliadin peptides spanning all unique 12mers contained within all Genbank entries described as wheat gliadins found in September 2001. This Pepset library was designed “manually” from gene-derived protein sequences aligned using ClustalW software (MegAlign) arranged into phylogenetic groupings.

[0212]Approximately 0.6 micromole of each of 652 of the 20mers was provided. Two marker 20mer peptides were included in each set of 96 (VLQQHNIAHGSSQVLQESTY—peptide 161, and I...

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Abstract

The invention herein disclosed is related to epitopes useful in methods of diagnosing, treating, and preventing coeliac disease. Therapeutic compositions which comprise at least one epitope are provided.

Description

[0001]The invention relates to epitopes useful in the diagnosis and therapy of coeliac disease, including diagnostics, therapeutics, kits, and methods of using the foregoing.[0002]Coeliac disease is caused by an immune mediated hypersensitivity to dietary gluten. Gluten proteins in wheat, rye, barley and in some cases oats are toxic in coeliac disease. Gluten is composed of alpha / beta, gamma and omega gliadins, and low and high molecular weight (LMW and HMW) glutenins in wheat, hordeins in barley, secalins in rye and avenins in oats. Hordeins and secalins are homologous to gamma and omega gliadins and low and high molecular weight glutenins in wheat. Avenins are phylogenetically more distant than hordeins and secalins from wheat gluten.[0003]The goal of research in coeliac disease has been to define the toxic components of gluten by defining the peptides that stimulate gluten-specific T-cells. Precise definition of gluten epitopes permits development of new diagnostics, therapeutics...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/00C07K14/00C12N15/11C12N15/00C12N5/06C12N1/20C12N5/04C12P21/04A61P1/00A23L7/10A61K38/10A61K38/16C07K14/415C12N15/29G01N33/53
CPCA61K39/00A61K38/168A61P1/00A61P37/00A61P37/04A61P37/06A61P37/08
Inventor ANDERSON, ROBERTBEISSBATH, TIMTYE-DIN, JASON
Owner BTG INT LTD
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