Toll-like receptor 9 is involved in the induction of galectin-9 protein by dietary anti-allergic compound fucoidan

Gnagnan J. E. Ezan; Masashi Mizuno; Gnagnan J. E. Ezan; Masashi Mizuno

doi:10.3934/Allergy.2023002

AIMS Allergy and Immunology

2023, Volume 7, Issue 1: 24-39. doi: 10.3934/Allergy.2023002

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Research article

Toll-like receptor 9 is involved in the induction of galectin-9 protein by dietary anti-allergic compound fucoidan

Gnagnan J. E. Ezan ,
Masashi Mizuno ^,

Department of Agrobioscience, Graduate School of Agricultural Science, Kobe University, Kobe, Japan

Received: 24 October 2022 Revised: 16 November 2022 Accepted: 02 December 2022 Published: 04 January 2023

Dietary intervention of fucoidan extracted from Saccharina japonica brown seaweed has been ascertained to favor an increase of galectin-9 protein in the intestine of allergic mice, resulting in the attenuation of the food allergy symptoms. The molecular mechanism underpinning that galectin-9 secretion remains unclear. Recently, some evidence has suggested an implication of Toll-like receptor 9 (TLR9) in galectin-9 secretion. However, no investigation has been done. For this study, we aimed to understand the relationship between galectin-9 production and fucoidan intake, which will improve the therapeutic use of fucoidan in allergy treatment. Intestinal epithelial cells (IECs) were cultured in solid or transwell plates and apically exposed to fucoidan solutions and/or synthetic TLR9 agonist (CpG-ODN). The transcriptional response of the cells to galectin-9 (lgals9) and the TLR9 gene was evaluated by using q-RTPCR, and the protein expression of galectin-9 was analyzed by conducting an ELISA test. Knockdown of TLR9 in IECs was performed by targeting TLR9 siRNA, and its effect on galectin-9 release was assessed. We found that the interaction of fucoidan and IECs resulted in the upregulation of galectin-9 released in a dose- and time-dependent manner. The increase was further potentiated in combination with the TLR9 agonist. Fucoidan exposure to IECs tended to increase the mRNA expression of TLR9 in a way similar to that of the TLR9 agonist effect, and knockdown of TLR9 in IECs resulted in a decreased tendency of fucoidan-induced galectin-9 protein. TLR9 activation is therefore involved in the increased release of galectin-9 protein observed in IECs upon fucoidan exposure.
- food allergy,
- dietary fucoidan,
- galectin-9,
- intestinal epithelial cells,
- in vitro study,
- Toll-like receptor 9
Citation: Gnagnan J. E. Ezan, Masashi Mizuno. Toll-like receptor 9 is involved in the induction of galectin-9 protein by dietary anti-allergic compound fucoidan[J]. AIMS Allergy and Immunology, 2023, 7(1): 24-39. doi: 10.3934/Allergy.2023002

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Abstract

Dietary intervention of fucoidan extracted from Saccharina japonica brown seaweed has been ascertained to favor an increase of galectin-9 protein in the intestine of allergic mice, resulting in the attenuation of the food allergy symptoms. The molecular mechanism underpinning that galectin-9 secretion remains unclear. Recently, some evidence has suggested an implication of Toll-like receptor 9 (TLR9) in galectin-9 secretion. However, no investigation has been done. For this study, we aimed to understand the relationship between galectin-9 production and fucoidan intake, which will improve the therapeutic use of fucoidan in allergy treatment. Intestinal epithelial cells (IECs) were cultured in solid or transwell plates and apically exposed to fucoidan solutions and/or synthetic TLR9 agonist (CpG-ODN). The transcriptional response of the cells to galectin-9 (lgals9) and the TLR9 gene was evaluated by using q-RTPCR, and the protein expression of galectin-9 was analyzed by conducting an ELISA test. Knockdown of TLR9 in IECs was performed by targeting TLR9 siRNA, and its effect on galectin-9 release was assessed. We found that the interaction of fucoidan and IECs resulted in the upregulation of galectin-9 released in a dose- and time-dependent manner. The increase was further potentiated in combination with the TLR9 agonist. Fucoidan exposure to IECs tended to increase the mRNA expression of TLR9 in a way similar to that of the TLR9 agonist effect, and knockdown of TLR9 in IECs resulted in a decreased tendency of fucoidan-induced galectin-9 protein. TLR9 activation is therefore involved in the increased release of galectin-9 protein observed in IECs upon fucoidan exposure.

Acknowledgments

The study was supported by the Ministry of Education, Culture, Sports, Science and Technology of Japan through the MEXT scholarship program and JSPS KAKENHI Grant Number JP18K19741.

Conflict of interest

All authors declare no conflicts of interest regarding the publication of this paper.

Author contributions

Conceptualization: E.E and M.M; Methodology: E.E and M.M; Manuscript draft: E.E; Manuscript review and approval: E.E and M.M.

References

[1]	Li B, Lu F, Wei X, et al. (2008) Fucoidan: structure and bioactivity. Molecules 13: 1671-1695. https://doi.org/10.3390/molecules13081671
[2]	Dobrinčić A, Balbino S, Zorić Z, et al. (2020) Advanced technologies for the extraction of marine brown algal polysaccharides. Mar Drugs 18: 168. https://doi.org/10.3390/md18030168
[3]	Ale MT, Meyer AS (2013) Fucoidans from brown seaweeds: an update on structures, extraction techniques and use of enzymes as tools for structural elucidation. RSC Adv 3: 8131-8141. https://doi.org/10.1039/C3RA23373A
[4]	Liu J, Wu SY, Chen L, et al. (2020) Different extraction methods bring about distinct physicochemical properties and antioxidant activities of Sargassum fusiforme fucoidans. Int J Biol Macromol 155: 1385-1392. https://doi.org/10.1016/j.ijbiomac.2019.11.113
[5]	Ale MT, Mikkelsen JD, Meyer AS (2011) Important determinants for fucoidan bioactivity: A critical review of structure-function relations and extraction methods for fucose-containing sulfated polysaccharides from brown seaweeds. Mar Drugs 9: 2106-2130. https://doi.org/10.3390/md9102106
[6]	Zhu Q, Chen J, Li Q, et al. (2016) Antitumor activity of polysaccharide from Laminaria japonica on mice bearing H22 liver cancer. Int J Biol Macromol 92: 156-158. https://doi.org/10.1016/j.ijbiomac.2016.06.090
[7]	Peng Y, Song Y, Wang Q, et al. (2019) In vitro and in vivo immunomodulatory effects of fucoidan compound agents. Int J Biol Macromol 127: 48-56. https://doi.org/10.1016/j.ijbiomac.2018.12.197
[8]	Luan F, Zou J, Rao Z (2021) Polysaccharides from Laminaria japonica: an insight into the current research on structural features and biological properties. Food Funct 12: 4254-4283. https://doi.org/10.1039/D1FO00311A
[9]	Herath KHINM, Kim HJ, Kim A, et al. (2020) The role of fucoidans isolated from the sporophylls of undaria pinnatifida against particulate-matter-induced allergic airway inflammation: Evidence of the attenuation of oxidative stress and inflammatory responses. Molecules 25: 2869. https://doi.org/10.3390/molecules25122869
[10]	Yang CH, Tian JJ, Ko WS, et al. (2019) Oligo-fucoidan improved unbalance the Th1/Th2 and Treg/Th17 ratios in asthmatic patients: An ex vivo study. Exp Ther Med 17: 3-10. https://doi.org/10.3892/etm.2018.6939
[11]	Tomori M, Nagamine T, Miyamoto T, et al. (2019) Evaluation of the immunomodulatory effects of fucoidan derived from Cladosiphon okamuranus Tokida in mice. Mar Drugs 17: 547. https://doi.org/10.3390/md17100547
[12]	Yanase Y, Hiragun T, Uchida K, et al. (2009) Peritoneal injection of fucoidan suppresses the increase of plasma IgE induced by OVA-sensitization. Biochem Biophys Res Commun 387: 435-439. https://doi.org/10.1016/j.bbrc.2009.07.031
[13]	Vo TS (2020) The role of algal fucoidans in potential anti-allergic therapeutics. Int J Biol Macromol 165: 1093-1098. https://doi.org/10.1016/j.ijbiomac.2020.09.252
[14]	Tanino Y, Hashimoto T, Ojima T, et al. (2016) F-fucoidan from Saccharina japonica is a novel inducer of galectin-9 and exhibits anti-allergic activity. J Clin Biochem Nutr 59: 25-30. https://doi.org/10.3164/jcbn.15-144
[15]	Mizuno M, Sakaguchi K, Sakane I (2020) Oral administration of fucoidan can exert anti-allergic activity after allergen sensitization by enhancement of galectin-9 secretion in blood. Biomolecules 10: 258. https://doi.org/10.3390/biom10020258
[16]	Niki T, Tsutsui S, Hirose S, et al. (2009) Galectin-9 is a high affinity ige-binding lectin with anti-allergic effect by blocking ige-antigen complex formation. J Biol Chem 284: 32344-32352. https://doi.org/10.1074/jbc.M109.035196
[17]	Nagamine N, Nakazato K, Tomioka S, et al. (2015) Intestinal absorption of fucoidan extracted from the brown seaweed, Cladosiphon okamuranus. Mar Drugs 13: 48-64. https://doi.org/10.3390/md13010048
[18]	Tokita Y, Nakajima K, Mochida H, et al. (2010) Development of a fucoidan-specific antibody and measurement of fucoidan in serum and urine by sandwich ELISA. Biosci Biotechnol Biochem 74: 350-357. https://doi.org/10.1271/bbb.90705
[19]	de Kivit S, Saeland E, Kraneveld AD, et al. (2012) Galectin-9 induced by dietary synbiotics is involved in suppression of allergic symptoms in mice and humans. Allergy 67: 343-352. https://doi.org/10.1111/j.1398-9995.2011.02771.x
[20]	de Kivit S, Kraneveld AD, Knippels LMJ, et al. (2013) Intestinal epithelium-derived galectin-9 is involved in the immunomodulating effects of nondigestible oligosaccharides. J Innate Immun 5: 625-638. https://doi.org/10.1159/000350515
[21]	de Kivit S, Kostadinova AI, Kerperien J, et al. (2017) Galectin-9 produced by intestinal epithelial cells enhances aldehyde dehydrogenase activity in dendritic cells in a PI3K- and p38-dependent manner. J Innate Immun 9: 609-620. https://doi.org/10.1159/000479817
[22]	Overbeek SA, Kostadinova AI, Boks MA, et al. (2019) Combined exposure of activated intestinal epithelial cells to nondigestible oligosaccharides and CpG-ODN suppresses Th2-associated CCL22 release while enhancing galectin-9, TGFβ, and Th1 polarization. Mediators Inflamm 2019: 8456829. https://doi.org/10.1155/2019/8456829
[23]	Ayechu-Muruzabal V, Overbeek SA, Kostadinova AI, et al. (2020) Exposure of intestinal epithelial cells to 2′-fucosyllactose and CpG enhances galectin release and instructs dendritic cells to drive Th1 and regulatory-type immune development. Biomolecules 10: 784. https://doi.org/10.3390/biom10050784
[24]	Ayechu-Muruzabal V, van de Kaa M, Mukherjee R, et al. (2022) Modulation of the epithelial-immune cell crosstalk and related galectin secretion by DP3-5 galacto-oligosaccharides and β-3′galactosyllactose. Biomolecules 12: 384. https://doi.org/10.3390/biom12030384
[25]	Lee J, Mo JH, Katakura K, et al. (2006) Maintenance of colonic homeostasis by distinctive apical TLR9 signaling in intestinal epithelial cells. Nat Cell Biol 8: 1327-1336. https://doi.org/10.1038/ncb1500
[26]	Ewaschuk JB, Backer JL, Churchill TA, et al. (2007) Surface expression of Toll-like receptor 9 is upregulated on intestinal epithelial cells in response to pathogenic bacterial DNA. Infect Immun 75: 2572-2579. https://doi.org/10.1128/IAI.01662-06
[27]	Eaton-Bassiri A, Dillon SB, Cunningham M, et al. (2004) Toll-like receptor 9 can be expressed at the cell surface of distinct populations of tonsils and human peripheral blood mononuclear cells. Infect Immun 72: 7202-7211. https://doi.org/10.1128/IAI.72.12.7202-7211.2004
[28]	Agier J, Żelechowska P, Kozłowska E, et al. (2016) Expression of surface and intracellular Toll-like receptors by mature mast cells. Cent Eur J Immunol 41: 333-338. https://doi.org/10.5114/ceji.2016.65131
[29]	Onji M, Kanno A, Saitoh SI, et al. (2013) An essential role for the N-terminal fragment of Toll-like receptor 9 in DNA sensing. Nat Com 4: 1949. https://doi.org/10.1038/ncomms2949
[30]	Jin JO, Park HY, Xu Q, et al. (2009) Ligand of scavenger receptor class A indirectly induces maturation of human blood dendritic cells via production of tumor necrosis factor-α. Blood 113: 5839-5847. https://doi.org/10.1182/blood-2008-10-184796
[31]	Lin Z, Tan X, Zhang Y, et al. (2020) Molecular targets and related biologic activities of fucoidan: A review. Mar Drugs 18: 376. https://doi.org/10.3390/md18080376
[32]	Choi S, Jeon SA, Heo BY, et al. (2022) Gene set enrichment analysis reveals that fucoidan induces type I IFN pathways in BMDC. Nutrients 14: 2242. https://doi.org/10.3390/nu14112242
[33]	Yamazaki Y, Nakamura Y, Nakamura T (2016) A fluorometric assay for quantification of fucoidan, a sulfated polysaccharide from brown algae. Plant Biotechnol J 33: 117-121. https://doi.org/10.5511/plantbiotechnology.16.0217c
[34]	Makarenkovaa ID, Logunov DY, Tukhvatulin AI, et al. (2012) Sulfated polysaccharides of brown seaweeds are ligands of Toll-like receptors. Bio Chem 6: 75-80. https://doi.org/10.1134/S1990750812010118
[35]	Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63. https://doi.org/10.1016/0022-1759(83)90303-4
[36]	Ghadimi D, de Vrese M, Heller KJ, et al. (2010) Effect of natural commensal-origin DNA on Toll-like receptor 9 (TLR9) signaling cascade, chemokine IL-8 expression, and barrier integrity of polarized intestinal epithelial cells. Inflamm Bowel Dis 16: 410-427. https://doi.org/10.1002/ibd.21057
[37]	Zhang X, Wei Z, Xue C (2021) Physicochemical properties of fucoidan and its applications as building blocks of nutraceutical delivery systems. Crit Rev Food Sci Nutr 2021: 1-19.
[38]	Ricós-Muñoz N, Maicas S, Pina-Pérez MC (2021) Probiotic Lactobacillus reuteri growth improved under fucoidan exposure. Proceedings 70: 106. https://doi.org/10.3390/foods_2020-07724
[39]	Shang Q, Shan X, Cai C, et al. (2016) Dietary fucoidan modulates the gut microbiota in mice by increasing the abundance of Lactobacillus and Ruminococcaceae. Food Funct 7: 3224-3232. https://doi.org/10.1039/C6FO00309E
[40]	Hemmi H, Takeuchi O, Kawai T, et al. (2000) A Toll-like receptor recognizes bacterial DNA. Nature 408: 740-745. https://doi.org/10.1038/35047123
[41]	Schneberger D, Caldwell S, Kanthan R, et al. (2013) Expression of Toll-like receptor 9 in mouse and human lungs. J Anat 222: 495-503. https://doi.org/10.1111/joa.12039
[42]	Leifer CA, Kennedy MN, Mazzoni A, et al. (2004) TLR9 is localized in the endoplasmic reticulum prior to stimulation. J Immunol 173: 1179-1183. https://doi.org/10.4049/jimmunol.173.2.1179
[43]	Latz E, Schoenemeyer A, Visintin A, et al. (2004) TLR9 signals after translocating from the ER to CpG DNA in the lysosome. Nat Immunol 5: 190-198. https://doi.org/10.1038/ni1028
[44]	Varga MG, Lin HC (2020) The role of toll-like receptor 9 in maintaining gut homeostasis. Ann Syst Biol 3: 10-14. https://doi.org/10.17352/asb.000005
[45]	Mielcarska MB, Bossowska-Nowicka M, Toka FN (2021) Cell surface expression of endosomal toll-like receptors—A necessity or a superfluous duplication?. Front Immunol 11: 620972. https://doi.org/10.3389/fimmu.2020.620972

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