1. Structural characteristics, immune-activating mechanisms in vitro, and immunomodulatory effects in vivo of the exopolysaccharide EPS53 from Streptococcus thermophilus XJ53.
- Author
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Yue Y, Han J, Shen X, Zhu F, Liu Y, Zhang W, Xia W, and Wu M
- Subjects
- Animals, Mice, RAW 264.7 Cells, Toll-Like Receptor 4 metabolism, NF-kappa B metabolism, Reactive Oxygen Species metabolism, Macrophages drug effects, Macrophages metabolism, Cytokines metabolism, Immunologic Factors pharmacology, Immunologic Factors chemistry, Immunomodulating Agents pharmacology, Immunomodulating Agents chemistry, Signal Transduction drug effects, Streptococcus thermophilus, Zebrafish, Polysaccharides, Bacterial chemistry, Polysaccharides, Bacterial pharmacology, Phagocytosis drug effects
- Abstract
Our previous investigations have successfully identified the repeating structural units of EPS53, an exopolysaccharide derived from Streptococcus thermophilus XJ53 fermented milk, and substantiated its potential immunomodulatory properties. The present study further elucidated the structural characteristics of EPS53 and investigated the underlying mechanisms governing its in vitro immunoreactivity as well as its in vivo immunoreactivity. The results obtained from multi-detector high performance gel filtration chromatography revealed that EPS53 adopted a rigid rod conformation in aqueous solution, with the weight-average molecular weight of 1464 kDa, the number-average molecular weight of 694 kDa, and the polydispersity index of 2.11. Congo red experiment confirmed the absence of a triple helix conformation. Scanning electron microscopy showed that EPS53 displayed a three-dimensional fibrous structure covered with flakes. The in vitro findings indicated that EPS53 enhanced phagocytosis ability, reactive oxygen species (ROS) production, and cytokine levels of macrophages via the TLR4-mediated NF-κB/MAPK signaling pathways as confirmed by immunofluorescence staining experiments, inhibition blocking experiments, and Western blot assay. Additionally, the in vivo experiments demonstrated that EPS53 significantly increased macrophage and neutrophil number while enhancing NO and ROS levels in zebrafish larvae; thus, providing further evidence for the immunomodulatory efficacy of EPS53., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)
- Published
- 2024
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