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Published on: May 12, 2020
Effect of malondialdehyde on the ovalbumin structure and its interactions with T84 epithelial cells
Jasna Nikolić1, Andrijana Nešić1, Milena Čavić2
1Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Belgrade, Serbia.
Background:
Protein oxidation can occur as a consequence of lipid peroxidation during food processing. The aim of this work was to explore the effect of malondialdehyde (MDA) modification of ovalbumin (OVA) on its interaction with T84 intestinal cells.
Methods:
Molecular dynamics simulation was employed for the prediction of MDA modification in the OVA, while introduced structural changes were evaluated by measurement of carbonyl group content, fluorescence spectra, MS/MS analysis, and IgE reactivity. Effects of MDA modified OVA on T84 epithelial cells were analyzed by gene expression for pro-inflammatory cytokines and protein secretion.
Results:
Out of 9 predicted, five modified Lys residues were confirmed by MS/MS analysis: 51TQINKVVR58, 85DILNQITKPNDVYSFSLASR104, 111YPILPEYLQCVKELYR126, 187AFKDEDTQAMPFR199, 277KIKVYLPR284, and 278IKVYLPR284. The introduced MDA modifications influenced profile of IgE reactivity to OVA. Treatment of T84 epithelial cells with OVA and OVA modified with 1mM MDA, induced up-regulation of pro-inflammatory cytokines (IL-1β, IL-25, IL-33, TSLP and TNFα), while OVA modification with 10mM MDA induced down regulation of the cytokine expression profile, except for IL-1β. OVA and OVA modified with 1mM MDA induced secretion of epithelial cells specific cytokine IL-33.
Conclusions:
This finding indicated that OVA and its MDA modified form have the potential to trigger the innate immunity by inducing up-regulation and secretion of pro-allergenic IL-33 in T84 intestinal epithelial cells.
General Significance:
Interactions of ovalbumin and its MDA modified form with intestinal epithelial cells can induce a specific immunological priming necessary for the downstream activation of innate immunity.
Insights
Malondialdehyde modification of ovalbumin (OVA) impacts its interaction with intestinal cells. MDA-modified OVA can trigger innate immunity by up-regulating pro-allergenic cytokine IL-33 in T84 cells.
Area of Science:
- Food science and immunology
- Protein modification and cellular response
Background:
- Protein oxidation, a result of lipid peroxidation during food processing, can alter protein structure and function.
- Ovalbumin (OVA), a common food protein, can undergo modification by malondialdehyde (MDA).
Purpose of the Study:
- To investigate the effect of malondialdehyde (MDA) modification of ovalbumin (OVA) on its interaction with T84 intestinal cells.
- To understand the immunological consequences of MDA-induced OVA modification.
Main Methods:
- Molecular dynamics simulations predicted MDA modification sites on OVA.
- Structural changes were assessed via carbonyl content, fluorescence spectra, MS/MS analysis, and IgE reactivity.
- T84 intestinal cells were analyzed for gene expression of pro-inflammatory cytokines and protein secretion after OVA treatment.
Main Results:
- Five lysine residues in OVA were confirmed as modification sites by MS/MS analysis.
- MDA modification altered OVA's IgE reactivity profile.
- Treatment with OVA and 1mM MDA-OVA upregulated pro-inflammatory cytokines (IL-1β, IL-25, IL-33, TSLP, TNFα) in T84 cells.
- 10mM MDA-OVA downregulated most cytokines, except IL-1β.
- OVA and 1mM MDA-OVA induced secretion of IL-33 from T84 cells.
Conclusions:
- OVA and its MDA-modified forms can activate innate immunity.
- This activation involves the upregulation and secretion of the pro-allergenic cytokine IL-33 in T84 intestinal epithelial cells.
- These interactions prime the immune system for downstream innate immunity activation.
