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Gluten derived from wheat and related triticeae cereals possesses distinct amino acid sequences that provoke the immunopathogenic features of celiac disease (CD) in genetically susceptible individuals. However, the role of oat-derived gluten, or avenins, in CD pathogenesis remains a disputed matter, as evidenced by a lack in harmonized legislation regarding gluten classification in relation to gluten-free labeling. In this study, we have analyzed a panel of pure oat cultivars using a sandwich ELISA based on the R5 monoclonal antibody (mAb), which binds to canonical epitopes occurring within celiagenic peptides present in triticeae-derived gluten but reportedly not present in avenins. We have identified three varieties of oats that reproducibly bind R5 antibodies and levels indicating the presence of gluten at more than the 20 ppm gluten regulatory threshold. Nested assessment using Western blot analysis and alternative gluten detection systems corroborated these results. Collectively, these data suggest that select oat varieties may prove problematic to patients with CD and to food companies and regulatory agencies and will extend our basic understanding of current gluten detection systems.

Gluten derived from wheat and related Triticeae can induce gluten sensitivity as well as celiac disease. Consequently, gluten content in foods labeled "gluten-free" is regulated. Determination of potential contamination in such foods is achieved using immunoassays based on monoclonal antibodies (mAbs) that recognize specific epitopes present in gluten. However, food-processing measures can affect epitope recognition. In particular, preparation of wheat protein isolate through deamidation of glutamine residues significantly limits the ability of commercial gluten testing kits in their ability to recognize gluten. Adding to this concern, evidence suggests that deamidated gluten imparts more pathogenic potential in celiac disease than native gluten. To address the heightened need for antibody-based tools that can recognize deamidated gluten, we have generated a novel mAb, 2B9, and subsequently developed it as a rapid lateral flow immunoassay. Herein, we report the ability of the 2B9-based lateral flow device (LFD) to detect gluten from wheat, barley, and rye and deamidated gluten down to 2 ppm in food as well as its performance in food testing.

Cucumber mosaic virus Fast New York strain (CMV-Fny) containing a mutated 2b protein (CMV-FnyΔ2b) was evaluated for the ability to infect ‘Calwonder’ bell pepper (Capsicum annuum) plants in comparative tests with the parent virus, CMV-Fny. Plants inoculated with CMV-FnyΔ2b did not develop local or systemic symptoms of infection, whereas CMV-Fny-infected plants developed systemic chlorosis by 7 days post inoculation (dpi), followed by mosaic and leaf deformation. Virus accumulation, determined by enzyme-linked immunosorbent assay (ELISA), revealed that CMV-FnyΔ2b accumulated in inoculated Calwonder leaves and inconsistently infected some noninoculated leaves at a low titer but was not detected in the youngest, noninoculated leaves. Immuno-tissue blot tests did not detect CMV-FnyΔ2b in the stems of infected plants, whereas CMV-Fny accumulated throughout the length of the stems of inoculated plants. In two experiments, protoplasts were isolated from Calwonder leaves, inoculated with viral RNAs of CMV-Fny or CMV-FnyΔ2b, and tested by ELISA for infection. In both experiments, less CMV-FnyΔ2b than CMV-Fny accumulated in protoplasts. These results suggest that the CMV 2b protein is needed for systemic infection of Calwonder pepper plants and for accumulation of the virus in inoculated protoplasts.

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