BACKGROUND: Peanut allergy is one of the most severe food allergies. Whether breastfeeding induces tolerance to peanuts or on the contrary, pre-disposes at risk-babies to occult allergic sensitization to peanuts is still a matter of discussion. We sought to investigate the transfer of the most potent peanut allergen Ara h 2 into human breast milk in a German breast milk study and to shed light on the time kinetics of Ara h 2 appearance. METHODS: We recruited 32 lactating, non-peanut-allergic women and collected breast milk samples at different time points after consumption of 100 g dry roasted peanuts. Breast milk samples were investigated for Ara h 2 with different immunological methods: by 2D immunoblotting with a patient's serum, by affinity enrichment using a monoclonal antibody against Ara h 2 followed by LC-MS/MS-based detection and by a competitive inhibition ELISA for the detection of Ara h 2 and its digestion-resistant peptides (DRP-Ara h 2). RESULTS: In a qualitative analysis, Ara h 2 could be identified in a breast milk sample by 2D immunoblot by means of a patient's serum and furthermore by immunoaffinity enrichment followed by LC-MS/MS analysis. In a semi-quantitative analysis, Ara h 2 and its digestion-resistant peptides were detected in the breast milk of 9 of 32 subjects. Evidence suggests that Ara h 2 is excreted individually either rapidly (after 1, 2, 3 or 4 h) or delayed (after 8 or 12 h) and in different concentrations. CONCLUSIONS: Time and concentration of secreted Ara h 2 in breast milk appears to be individually regulated. The identification of Ara h 2 in breast milk is the prerequisite for the investigation of its sensitizing or tolerogenic properties.
- Schocker, F.
- Baumert, J.
- Kull, S.
- Petersen, A.
- Becker, W. M.
- Jappe, U.
Keywords
- 2S Albumins, Plant/immunology/*metabolism
- Adult
- Antigens, Plant/immunology/*metabolism
- Arachis/*immunology
- Blotting, Western
- *Breast Feeding
- Chromatography, Liquid
- Enzyme-Linked Immunosorbent Assay
- Female
- Germany
- Glycoproteins/immunology/*metabolism
- Humans
- Kinetics
- Milk, Human/immunology/*metabolism
- Nuts/*immunology
- Peanut Hypersensitivity/*immunology
- Prospective Studies
- Tandem Mass Spectrometry
- Ara h 2
- Elisa
- allergen transfer
- digestion-resistant peptides of Ara h 2
- human breast milk
- immunoaffinity enrichment
- immunoblot analysis