Abstract
There is a need to assess the relationship between improved rheological properties and the immunogenic potential of wheat proteins. The present study aimed to investigate the in vitro effects of total protein extracts from three modern and two landrace Triticum aestivum commercial flour mixes, with significant differences in gluten strength (GS), on cell lines. Cytotoxicity and innate immune responses induced by wheat proteins were investigated using Caco-2 monocultures, two dimensional (2D) Caco-2/U937 co-cultures, and three dimensional (3D) co-cultures simulating the intestinal mucosa with Caco-2 epithelial cells situated above an extra-cellular matrix containing U937 monocytes and L929 fibroblasts. Modern wheat proteins, with increased GS, significantly reduced Caco-2 cell proliferation and vitality in monoculture and 2D co-cultures than landrace proteins. Modern wheat proteins also augmented Caco-2 monolayer disruption and tight junction protein, occludin, redistribution in 3D co-cultures. Release of interleukin-8 into the cell medium and increased U937 monocyte migration in both 2D and 3D co-cultures were similarly apparent. Immuno-activation of migrating U937 cells was evidenced from cluster of differentiation 14 (CD14) staining and CD11b-related differentiation into macrophages. The modern wheat proteins, with gluten polymorphism relatedness and increased GS, were shown to be more cytotoxic and immunogenic than the landrace wheat proteins.
Highlights
Wheat is one of the major staple crops worldwide, with approximately 95% of total production being represented by Triticum aestivum L. ssp. aestivum, generally referred to as common wheat
Combining the requirement to examine the immunogenic potential of wheat genotypes on the basis of improved technological parameters together with that of expanding comparative studies between heritage and modern genotypes, the present study adopted a function-based approach by using cell lines to measure the effects T. aestivum protein extracts from commercially available wheat sources of modern and heritage flours in relation to gluten strength (GS)
Flour mixes with increased GS were more cytotoxic, resulting in a greater proinflammatory-induced reduction in cell proliferation of Caco-2 cells in monoculture, as well as in the vitality of Caco-2 cells and migration of U937 cells in 2D co-culture. These results were supported by 3D co-cultures, showing that the higher GS proteins induced a greater damage to Caco-2 monolayer integrity, increased tight junction (TJ) protein delocalization, as well as an increased activation U937 monocytes, and resultant differentiation into macrophages, compared to the heritage flour mixes
Summary
The seed storage proteins (8–15% of the total flour weight) are important determinants of end-product quality, with improved bread-making properties largely attributable to the D genome [1,2]. Potent reactive components in WA (Baker’s asthma, atopic dermatitis, urticaria, and anaphylaxis) include α- and ω-gliadins, glutenins, as well as amylase-trypsin inhibitors (ATIs), serpins, and wheat lipid transfer proteins associated with the albumin/globulin fraction [1]. Faced with a rapidly evolving public health issue in the form of wheat-related disorders combined with the lack of scientific evidence regarding the higher immune-stimulatory potential of modern wheat cultivars [3], research focus has been directed toward the study of gluten proteins in old and modern genotypes of both T. aestivum and T. durum
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