In vivo Evidence of Cross-Reactivity between Cow’s Milk and Soybean Proteins in a Mouse Model of Food Allergy

Abstract

Background: Cow’s milk allergy (CMA) is an important problem worldwide and the development of an in vivo system to study new immunotherapeutic strategies is of interest. Intolerance to soybean formula has been described in CMA patients, but it is not fully understood. In this work, we used a food allergy model in BALB/c mice to study the cross-reactivity between cow’s milk protein (CMP) and soy proteins (SP). Methods: Mice were orally sensitized with cholera toxin and CMP, and then challenged with CMP or SP to induce allergy. Elicited symptoms, plasma histamine, humoral and cellular immune response were analyzed. Th1- and Th2-associated cytokines and transcription factors were assessed at mucosal sites and in splenocytes. Cutaneous tests were also performed. Results: We found that the immediate symptoms elicited in CMP-sensitized mice orally challenged with SP were consistent with a plasma histamine increase. The serum levels of CMP-specific IgE and IgG1 antibodies were increased. These antibodies also recognized soy proteins. Splenocytes and mesenteric lymph node cells incubated with CMP or SP secreted IL-5 and IL-13. mRNA expression of Th2-associated genes (IL-5, IL-13, and GATA-3) was upregulated in mucosal samples. In addition, sensitized animals exhibited positive cutaneous tests after the injection of CMP or SP. Conclusions: We demonstrate that CMP-sensitized mice, without previous exposure to soy proteins, elicited hypersensitivity signs immediately after the oral administration of SP, suggesting that the immunochemical cross-reactivity might be clinically relevant. This model may provide an approach to further characterize cross-allergenicity phenomena and develop new immunotherapeutic treatments for allergic patients.

Constipation and Cow's Milk Allergy: A Review of the Literature

“…[25] The same recommendation was reaffirmed in the updated document of 2008 [26]. Furthermore, it was recently shown that CM-sensitized mice without previous exposure to SP elicited hypersensitivity signs immediately after the oral administration of SP, suggesting that immunochemical cross-reactivity might be clinically relevant [27]. Consistent with these indications, Irastorza et al [14] did not allow their patients to ingest soy.…”

Oral delivery ofBrucellaspp. recombinant protein U-Omp16 abrogates the IgE-mediated milk allergy

“…33,12 Briefly, recombinant U-Omp16 was isolated from bacterial cytoplasm and then purified by affinity chromatography with a Ni-NTA resin (Qiagen). Expression and purification of the recombinant protein was checked by SDS-PAGE followed by Coomasie Blue staining and to confirm the identity of the U-Omp16, western blot was performed and developed with anti-Omp16-specific mAb (data not shown).…”

“…33 Briefly, mice received 6 weekly intragastric (i.g) doses of 20 mg of CMP administered as homogenized commercial non-fat dry milk, plus 10 μg of cholera toxin (CT) (Sigma Aldrich) in a final volume of 200 μL of bicarbonate buffer (sensitized mice). As control group naïve mice received 6 weekly i.g doses of 20 mg CMP with PBS.…”

“…33 Briefly, cyanogen bromide-activated cellulose paper discs were coupled with CMP extracts containing 1.75 mg/mL of protein.…”

“…33 Briefly, Maxi Sorp ELISA plates (NUNC) were coated overnight at 4 °C with 1 μg/100 μL CMP in carbonate-bicarbonate buffer (pH 9.6). Coated plates were blocked with 5% equine serum in saline for 2 h at 37 °C and then incubated with sera (1/200) for 1 h at 37 °C.…”