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T‐cell epitope‐containing hypoallergenic β‐lactoglobulin for oral immunotherapy in milk allergy
Author(s) -
Ueno Hiroshi M.,
Kato Teruhiko,
Ohnishi Hidenori,
Kawamoto Norio,
Kato Zenichiro,
Kaneko Hideo,
Kondo Naomi,
Nakano Taku
Publication year - 2016
Publication title -
pediatric allergy and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.269
H-Index - 89
eISSN - 1399-3038
pISSN - 0905-6157
DOI - 10.1111/pai.12642
Subject(s) - hypoallergenic , medicine , oral immunotherapy , epitope , allergy , immunotherapy , allergen , beta lactoglobulin , immunology , milk allergy , food allergy , antibody , whey protein , food science , immune system , chemistry
Abstract Background Optimally hydrolyzed β‐Lactoglobulin (βLg) is a promising milk oral immunotherapy (OIT) candidate with respect to showing reduced B‐cell reactivity but retaining the T‐cell epitope. To demonstrate that an edible hypoallergenic βLg hydrolysate containing the T‐cell epitope is suitable for OIT. We tested how chymotrypsin affected the retention of the T‐cell epitope of βLg when preparing βLg hydrolysates using food‐grade trypsin. Methods We investigated the effect of chymotrypsin activity on the formation of the T‐cell epitope‐containing peptide of βLg (βLg 102–124 ) and prepared an edible βLg hydrolysate containing βLg 102–124 using screened food‐grade trypsins. B‐cell reactivity was determined using immunoassays in which ELISA was performed with anti‐βLg rabbit IgG and Western blotting was performed with a milk‐specific IgE antiserum. Results In βLg hydrolysis performed by varying the activity of trypsin and chymotrypsin, chymotrypsin activity inhibited the formation of βLg 102–124 with an increase in hydrolysis time in a dose‐dependent manner. βLg 102–124 was generated by two of five food‐grade trypsins used at a ratio of 1:50 (w/w, enzyme/substrate) for 20 h at 40°C. The edible βLg hydrolysate retained βLg 102–124 and showed a reduction in molecular weight distribution and antigenicity against IgG and IgE. Conclusions Chymotrypsin activity inhibited the formation of βLg 102–124 in the trypsin hydrolysate of βLg. This βLg trypsin hydrolysate is a novel candidate for peptide‐based OIT in cow's milk allergy for safely inducing desensitization.