Effect of simulated gastro-duodenal digestion on the allergenic reactivity of beta-lactoglobulin

Clinical and Translational Allergy

Photini Papageorgiou⁴  Clare EN Mills⁵  Jean-Michel Wal¹  Karine Adel-Patient¹  Hervé Bernard¹  Nikolaos G Papadopoulos⁴  Neil M Rigby⁵  Lucie Mondoulet³  Maria Theodoropoulou⁶  Apostolos Bossios² 

[1] Lab d'Immuno-Allergie Alimentaire, INRA-CEA, Saclay, France;Krefting Research Centre, Department of Internal Medicine, Institute of Medicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden;DBV Technologies, 92220, Bagneux, France;Athens University, P. & A. Kyriakou Children's Hospital, Athens, Greece;Institute of Food Research, Norwich Research Park, Colney NR4 7UA UK;Thessaly University Hospital, Larissa, Greece

关键词: skin prick test;    Basophil activation;    flow cytometry;    β-lactoglobulin;    cow's milk allergy;    in vitro digestion;   

Others  :  794815
DOI  :  10.1186/2045-7022-1-6

 received in 2011-05-31, accepted in 2011-08-09,  发布年份 2011

【 摘 要 】

Background

Cow's milk (CM) allergy affects about 2% of infants. The allergenicity of dietary proteins, including those from CM, has been related to their digestibility although the generality of the link and its causality remains to be demonstrated. In this study we use an in vitro digestion system, to investigate the digestibility of β-lactoglobulin (blg) during gastrointestinal transit and to assess the impact of this process on blg allergenic reactivity in CM allergic children.

Methods

Blg digesta were prepared using an in vitro digestion protocol simulating either gastric digestion alone or followed by duodenal digestion with or without phosphatidylcholine (PC). Biochemical analysis of blg digesta was performed by SDS-PAGE and their concentration was measured by a sandwich ELISA. Assessment of their allergenic reactivity was done in vitro by EAST inhibition, specific basophil activation (basotest) and lymphocyte proliferation (PCNA-flow cytometry) assays using sera and cells from patients allergic to blg and in vivo by skin prick testing (SPT) of these patients.

Results

Blg was only broken down to smaller peptides after gastro-duodenal digestion although a sizeable amount of intact protein still remained. Digestion did not modify the IgE binding capacity of blg except for gastro-duodenal digestion performed in the absence of PC. These results are consistent with the quantity of intact blg remaining in the digesta. Overall both gastric and gastroduodenal digestion enhanced activation of sensitized basophils and proliferation of sensitized lymphocytes by blg. However, there was a tendency towards reduction in mean diameter of SPT following digestion, the PC alone during phase 1 digestion causing a significant increase in mean diameter.

Conclusions

Digestion did not reduce the allergenic reactivity of blg to a clinically insignificant extent, PC inhibiting digestion and thereby protecting blg allergenic reactivity. SPT reactivity was reduced compared to blg immunoreactivity in in vitro tests.

【 授权许可】

   
2011 Bossios et al; licensee BioMed Central Ltd.

【 图 表 】

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