Anti-betalactoglobulin IgG antibodies bind to a specific profile of epitopes when patients are allergic to cow's milk proteins

Background We demonstrated recently that mite-allergic patients differed from healthy controls in the specificity of their IgG antibodies towards mite antigens. Objective The present study investigates whether these discriminatory IgG responses could be associated with the expression and the evolution of clinical manifestations in allergy to cow's milk proteins. Methods Antibody specificity was evaluated by comparing IgG-binding to native bovine beta-lactoglobulin (nBLG) and its products of pepsin hydrolysis (dBLG) using a solid-phase enzyme-linked immunosorbent assay (ELISA). Antibody specificity was further investigated in competitive ELISA using streptavidin-biotin technology with purified IgG fractions from selected subjects and specific mouse monoclonals raised against BLG. Results IgG antibodies from CM-intolerant or allergic sera (n = 222) showed a higher degree of binding to nBLG than to dBLG, while control sera showed similar levels to both nBLG and dBLG (n = 99 children/65 adults). Sera from symptomatic patients, wether or not they contained IgE antibodies, demonstrated group-segregating capacities to compete with pooled purified IgG from each clinical class, and with selected murine anti-nBLG monoclonal antibodies for binding to n- and dBLG. Furthermore, this inhibitory capacity shifted dramatically in a small subset (n = 14) of children as they developed CM-tolerance. Conclusion The IgG responses to BLG of CM-intolerant or allergic patients are very different from those of healthy controls, being characterized not only by increased titres but also similar patterns of modified specificity, including a marked preference for conformational epitopes. Cross-competition experiments confirmed that the restricted specificity was clinically associated, appearing as an immunological signature, which allowed almost complete discrimination between patient groups. This phenomenon is a particularly promising diagnostic feature in this category of young patients where conventional tests usually only document the status of sensitization.