Antibody response reveal differences in oral tolerance to wheat and maize grain protein fractions

The influence of diet on humoral immune responses to gluten- and maize-derived proteins was examined using ELISA and protein blotting techniques. Mice raised on the maize-based (gluten-free) diet responded well to parenteral immunization with each of six gluten-derived protein preparations (whole gliadin, two omega-gliadin fractions, wheat salt-soluble proteins, a peptic-tryptic digest and a subtilisin digest of gluten), as serum antibody levels increased at least 300-fold in each case. In contrast, mice raised on the wheat-based diet responded poorly to immunization with either whole gliadin or omega-gliadin and were virtually non-responsive to enzymic digest of gluten. Diet had little effect on the magnitude of the antibody response to wheat salt-soluble proteins, with both groups showing a 300-fold increase in titre. Similarly, tolerance to alpha-zeins, the alcohol-soluble proteins of maize, did not occur on either diet. However, some oral tolerance was observed to maize glutelin. The specificity of the various antibody responses was then analysed by immunoblotting. Following immunization with gluten proteins or digests, antibodies from the maize-fed mice bound more or less equally to each of the main gliadin bands and to the glutenins while the mice on the wheat-based diet had antibody specific for omega-gliadin proteins. Serum antibodies from the maize-fed mice, immunized with either alpha-zein or maize glutelin, showed even labelling of the major maize endosperm proteins while antibodies from mice on the wheat diet showed strong labelling of the Mr 27,000 and 58,000 bands. These results show that diet influenced the specificity, as well as the magnitude of serum antibody responses to cereal proteins. In addition, oral tolerance appeared to affect the humoral response to some cereal proteins more than others. Both of these findings have important implications for our understanding of coeliac disease.