Retinoic acid prevents immunogenicity of milk lipocalin Bos d 5 through binding to its immunodominant T-cell epitope

Sci Rep. 2018 Jan 25;8(1):1598. doi: 10.1038/s41598-018-19883-0.

Abstract

The major cow's milk allergen Bos d 5 belongs to the lipocalin protein family, with an intramolecular pocket for hydrophobic ligands. We investigated whether Bos d 5 when loaded with the active vitamin A metabolite retinoic acid (RA), would elicit differential immune responses compared to the unloaded state. By in silico docking an affinity energy of -7.8 kcal/mol was calculated for RA into Bos d 5. Loading of RA to Bos d 5 could be achieved in vitro, as demonstrated by ANS displacement assay, but had no effect on serum IgE binding in tolerant or challenge-positive milk allergic children. Bioinformatic analysis revealed that RA binds to the immunodominant T-cell epitope region of Bos d 5. In accordance, Bos d 5 significantly suppressed the CD3+ CD4+ cell numbers, proliferative response and IL-10, IL-13 and IFN-γ secretion from stimulated human PBMCs only when complexed with RA. This phenomenon was neither associated with apoptosis of T-cells nor with the activation of Foxp3+ T-cells, but correlated likely with enhanced stability to lysosomal digestion due to a predicted overlap of Cathepsin S cleavage sites with the RA binding site. Taken together, proper loading of Bos d 5 with RA may suppress its immunogenicity and prevent its allergenicity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allergens / immunology*
  • Allergens / metabolism*
  • Animals
  • Cattle
  • Cell Proliferation / drug effects
  • Epitopes, T-Lymphocyte / metabolism*
  • Humans
  • Immunoglobulin E / metabolism
  • Immunologic Factors / metabolism*
  • Interferon-gamma / metabolism
  • Interleukin-10 / metabolism
  • Interleukin-13 / metabolism
  • Leukocytes, Mononuclear / immunology
  • Lipocalins / immunology*
  • Lipocalins / metabolism*
  • Lysosomes / metabolism
  • Molecular Docking Simulation
  • Protein Binding
  • Proteolysis
  • Tretinoin / metabolism*

Substances

  • Allergens
  • Bos d 5 allergen, Bos taurus
  • Epitopes, T-Lymphocyte
  • IL10 protein, human
  • Immunologic Factors
  • Interleukin-13
  • Lipocalins
  • Interleukin-10
  • Immunoglobulin E
  • Tretinoin
  • Interferon-gamma