Dissecting the T-cell response to hordeins in coeliac disease can develop barley with reduced immunotoxicity

作者：Tanner G J^*; Howitt C A; Forrester R I; Campbell P M; Tye Din J A; Anderson R P

来源：Alimentary Pharmacology and Therapeutics, 2010, 32(9): 1184-1191.

DOI：10.1111/j.1365-2036.2010.04452.x

P>Background
Wheat, rye and barley prolamins are toxic to patients with coeliac disease. Barley is diploid with pure inbred cultivars available, and is attractive for genetic approaches to detoxification.
Aim
To identify barley hordein fractions which activated the interferon-gamma (IFN-gamma) secreting peripheral blood T-cells from coeliac volunteers, and compare immunotoxicity of hordeins from experimental barley lines.
Methods
To reactivate a T-cell response to hordein, volunteers underwent a 3-day oral barley challenge. Peripheral blood mononuclear cells (PBMC) were isolated from twenty-one HLA DQ2+ patients with confirmed coeliac disease. IFN-gamma ELISpot assays enumerated T-cells activated by purified prolamins and positive controls.
Results
Hordein-specific T-cells were induced by oral barley challenge. All prolamin fractions were immunotoxic, but D- and C-hordeins were most active. Barley lines lacking B- and C-hordeins had a 5-fold reduced hordein-content, and immunotoxicity of hordein extracts were reduced 20-fold compared with wild-type barley.
Conclusions
In vivo oral barley challenge offers a convenient and rapid approach to test the immunotoxicity of small amounts of purified hordeins using fresh T-cells from patients in high throughput overnight assays. Barley lines that did not accumulate B- and C-hordeins were viable, yet had substantially reduced immunotoxicity. Creation of hordein-free barley may therefore be possible.

• 出版日期2010-11-1
• 单位CSIRO; 河北医科大学