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

Karin Hufnagl, Debajyoti Ghosh, Stefanie Wagner, Alessandro Fiocchi, Lamia Dahdah, Rodolfo Bianchini, Nina Braun, Ralf Steinborn, Martin Hofer, Marion Blaschitz, Georg A Roth, Gerlinde Hofstetter, Franziska Roth-Walter, Luis F Pacios, Erika Jensen-Jarolim

Research output: Contribution to journalArticle

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.

Original languageEnglish
Number of pages12
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - Jan 25 2018

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Lipocalins
Immunodominant Epitopes
T-Lymphocyte Epitopes
Tretinoin
Milk
cathepsin S
T-Lymphocytes
Interleukin-13
Computational Biology
Vitamin A
Computer Simulation
Interleukin-10
Allergens
Immunoglobulin E
Digestion
Cell Count
Binding Sites
Apoptosis
Ligands
Serum

Keywords

  • Allergens/immunology
  • 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
  • Lysosomes/metabolism
  • Molecular Docking Simulation
  • Protein Binding
  • Proteolysis
  • Tretinoin/metabolism

Cite this

Retinoic acid prevents immunogenicity of milk lipocalin Bos d 5 through binding to its immunodominant T-cell epitope. / Hufnagl, Karin; Ghosh, Debajyoti; Wagner, Stefanie; Fiocchi, Alessandro; Dahdah, Lamia; Bianchini, Rodolfo; Braun, Nina; Steinborn, Ralf; Hofer, Martin; Blaschitz, Marion; Roth, Georg A; Hofstetter, Gerlinde; Roth-Walter, Franziska; Pacios, Luis F; Jensen-Jarolim, Erika.

In: Scientific Reports, Vol. 8, No. 1, 25.01.2018.

Research output: Contribution to journalArticle

Hufnagl, K, Ghosh, D, Wagner, S, Fiocchi, A, Dahdah, L, Bianchini, R, Braun, N, Steinborn, R, Hofer, M, Blaschitz, M, Roth, GA, Hofstetter, G, Roth-Walter, F, Pacios, LF & Jensen-Jarolim, E 2018, 'Retinoic acid prevents immunogenicity of milk lipocalin Bos d 5 through binding to its immunodominant T-cell epitope', Scientific Reports, vol. 8, no. 1. https://doi.org/10.1038/s41598-018-19883-0
Hufnagl, Karin ; Ghosh, Debajyoti ; Wagner, Stefanie ; Fiocchi, Alessandro ; Dahdah, Lamia ; Bianchini, Rodolfo ; Braun, Nina ; Steinborn, Ralf ; Hofer, Martin ; Blaschitz, Marion ; Roth, Georg A ; Hofstetter, Gerlinde ; Roth-Walter, Franziska ; Pacios, Luis F ; Jensen-Jarolim, Erika. / Retinoic acid prevents immunogenicity of milk lipocalin Bos d 5 through binding to its immunodominant T-cell epitope. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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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.",
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T1 - Retinoic acid prevents immunogenicity of milk lipocalin Bos d 5 through binding to its immunodominant T-cell epitope

AU - Hufnagl, Karin

AU - Ghosh, Debajyoti

AU - Wagner, Stefanie

AU - Fiocchi, Alessandro

AU - Dahdah, Lamia

AU - Bianchini, Rodolfo

AU - Braun, Nina

AU - Steinborn, Ralf

AU - Hofer, Martin

AU - Blaschitz, Marion

AU - Roth, Georg A

AU - Hofstetter, Gerlinde

AU - Roth-Walter, Franziska

AU - Pacios, Luis F

AU - Jensen-Jarolim, Erika

PY - 2018/1/25

Y1 - 2018/1/25

N2 - 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.

AB - 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.

KW - Allergens/immunology

KW - Animals

KW - Cattle

KW - Cell Proliferation/drug effects

KW - Epitopes, T-Lymphocyte/metabolism

KW - Humans

KW - Immunoglobulin E/metabolism

KW - Immunologic Factors/metabolism

KW - Interferon-gamma/metabolism

KW - Interleukin-10/metabolism

KW - Interleukin-13/metabolism

KW - Leukocytes, Mononuclear/immunology

KW - Lipocalins/immunology

KW - Lysosomes/metabolism

KW - Molecular Docking Simulation

KW - Protein Binding

KW - Proteolysis

KW - Tretinoin/metabolism

U2 - 10.1038/s41598-018-19883-0

DO - 10.1038/s41598-018-19883-0

M3 - Article

VL - 8

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

ER -