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

Aliment Pharmacol Ther 2010; 32: 1184–1191 Summary 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‐γ (IFN‐γ) 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‐γ 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.