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. 2020 Feb 18;82(2):177-183.
doi: 10.1292/jvms.19-0222. Epub 2019 Dec 25.

Hydrolyzed diets may stimulate food-reactive lymphocytes in dogs

Kenichi Masuda  1 , Atsushi Sato  1 , Atsushi Tanaka  1 , Akiko Kumagai  1
Affiliations

Hydrolyzed diets may stimulate food-reactive lymphocytes in dogs

Kenichi Masuda et al. J Vet Med Sci. .

Abstract

Hydrolyzed proteins are often prescribed for dogs with food hypersensitivity in food elimination programs. However, the potential of these diets to stimulate lymphocyte-mediated hypersensitivity is currently unknown. In this study, two commercially available hydrolyzed diets for dogs, D-1 (Aminopeptide Formula Dry, Royal Canin Japon, Tokyo, Japan), and D-2 (Canine z/d Ultra Dry, Hill's-Colgate (Japan) Ltd., Tokyo, Japan), were analyzed to identify residual proteins or peptides, as well as activated helper T-lymphocyte reactions in dogs with suspected food hypersensitivity. Proteins and peptides with molecular weights >1 kDa (majority 1.5-3.5 kDa) were detected in both diet extracts with sodium dodecyl sulfate polyacrylamide gel electrophoresis, and size exclusion chromatography. When peripheral blood mononuclear cells (PBMC's) from 316 dogs with suspected food allergies were cultured with hydrolyzed diet extracts, flow cytometry analysis revealed detectable levels of CD25low helper T-lymphocytes stimulated by D-1 and D-2 in 91 of 316, (28.8%), and 75 of 316 (23.7%) samples, respectively. These data indicated that the extracts contained proteins or peptides large enough to activate the lymphocytes. The percentages of CD25low helper T-lymphocytes stimulated by D-1 and D-2 extracts increased to 38.7% and 29.6%, respectively, in 186 of the original 316 samples (186/316, 58.9%), also reactive to poultry-related antigens. Thus, both poultry-related antigens, and D-1 and D-2 diet extracts may activate helper T-lymphocytes. These results demonstrate that hydrolyzed diets may contain proteins that stimulate helper T-lymphocytes, and may not be effective for treating all dogs with food hypersensitivity.

Keywords: diet; dog; food allergy; hydrolyzation; lymphocyte.

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Conflict of interest statement

K. Masuda is the Chief Executive Officer and a stockholder of Animal Allergy Clinical Laboratories, Inc.

Figures

Fig. 1.
Fig. 1.
SDS-PAGE of D-1 (Lane 1), D-2 (Lane 2), and chicken meat (Lane 3) extracts with Coomassie brilliant blue (CBB) and silver staining. The total protein amount applied to each lane was 10 µg. Note that there were no visualized bands in Lane 1. SDS-PAGE, sodium dodecyl sulphate-polyacrylamide gel electrophoresis.
Fig. 2.
Fig. 2.
Size-exclusion chromatography of D-1 extracts. The assay was repeated with three different product lots. Similar wave patterns were detected in all product lots.
Fig. 3.
Fig. 3.
Size-exclusion chromatography of D-2 extracts. The assay was repeated with two different product lots. Similar wave patterns were detected in all product lots.
See this image and copyright information in PMC

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