Abstract
Ovine ruminal epithelial cells (ORECs) not only have a physical barrier function but also can secrete host defence peptides (HDPs), such as sheep β-defensin-1 (SBD-1). As a feed additive, Saccharomyces cerevisiae can enhance the host’s innate immunity. β-glucan, a cell wall component of Saccharomyces cerevisiae, can stimulate innate immune responses and trigger the up-regulation of SBD-1 in ORECs. The signaling mechanisms involved in β-glucan-induced SBD-1 expression are not completely understood. The aim of this study was to identify the receptors and intracellular pathways involved in the up-regulation of SBD-1 induced by β-glucan. ORECs were cultured, and the regulatory mechanisms of β-glucan-induced up-regulation of SBD-1 were detected using quantitative real-time PCR (qPCR), enzyme-linked immunosorbent assay (ELISA), and western blotting. TLR-2 and MyD88 knockdown or inhibition attenuated β-glucan-induced SBD-1 expression. We also showed that inhibition of MAPK and NF-κB pathways significantly reduced β-glucan-induced SBD-1 expression. These results demonstrate that β-glucan-induced SBD-1 expression is TLR-2-MyD88-dependent and may be regulated by both MAPK and NF-κB pathways. Since NF-κB inhibition had a greater effect on the down-regulation of β-glucan-induced SBD-1 expression, the NF-κB pathway may be the dominant signaling pathway involved in the regulation of defensin expression. Our studies demonstrate that β-glucan-induced SBD-1 expression is mediated through the TLR-2-MyD88-NF-κB/MAPK pathway. Our results would contribute to the understanding of immunological modulations in the gastrointestinal tract triggered by probiotic yeast cell wall components.
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Abbreviations
- ANOVA :
-
analysis of variance
- COX-2 :
-
cyclooxygenase-2
- ELISA :
-
enzyme-linked immunosorbent assay
- IFN-γ :
-
interferon-γ
- iNOS :
-
inducible nitric oxide synthase
- MAPKs :
-
mitogen-activated protein kinases
- MyD88 :
-
myeloid differentiation factor 88
- NF-κB :
-
nuclear factor kappa-light-chain-enhancer of activated B cells
- ORECs :
-
ovine ruminal epithelial cells
- PBS :
-
phosphate-buffered saline
- PCR :
-
polymerase chain reaction
- PRRs :
-
pattern recognition receptors
- qPCR :
-
quantitative real-time PCR
- SBD-1 :
-
sheep β-defensin-1
- SD :
-
standard deviation
- TLR-2 :
-
Toll-like receptor 2
- TLRs :
-
Toll-like receptors
- TNF-α :
-
tumor necrosis factor-α
References
Abdel-Mageed AM, Isobe N, Yoshimura Y (2014) Effects of different TLR ligands on the expression of proinflammatory cytokines and avian betadefensins in the uterine and vaginal tissues of laying hens. Vet Immunol Immunopathol 162(3–4):132–141
Aguilar-Uscanga B, Francois JM (2003) A study of the yeast cell wall composition and structure in response to growth conditions and mode of cultivation. Lett Appl Microbiol 37(3):268–274
Batbayar S, Kim MJ, Kim HW (2011) Medicinal mushroom Lingzhi or Reishi, Ganoderma lucidum (W.Curt.:Fr.) P. Karst., beta-glucan induces toll-like receptors and fails to induce inflammatory cytokines in NF-kappaB inhibitor-treated macrophages. Int J Med Mushrooms 13(3):213–225
Byeon SE, Lee J, Lee E, Lee SY, Hong EK, Kim YE, Cho JY (2009) Functional activation of macrophages, monocytes and splenic lymphocytes by polysaccharide fraction from Tricholoma matsutake. Arch Pharm Res 32(11):1565–1572
Byun EB, Park SH, Jang BS, Sung NY, Byun EH (2016) Gamma-irradiated β-glucan induces immunomodulation and anticancer activity through MAPK and NF-κB pathways. J Sci Food Agric 96(2):695–702
Campoverde C, Milne DJ, Estevez A, Duncan N, Secombes CJ, Andree KB (2017) Ontogeny and modulation after PAMPs stimulation of β-defensin, hepcidin, and piscidin antimicrobial peptides in meagre ( Argyrosomus regius ). Fish Shellfish Immunol 69:200–210
Chaucheyras-Durand F, Durand H (2010) Probiotics in animal nutrition and health. Benefic Microbes 1(1):3–9
Coyle C, Wheelhouse N, Jacques M, Longbottom D, Svoboda P, Pohl J, Duncan WC, Rae MT, Barlow PG (2016) Ovine trophoblasts express cathelicidin host defence peptide in response to infection. J Reprod Immunol 117:10–16
Czech A, Smolczyk A, Ognik K, Wlazło Ł, Nowakowicz-Dębek B, Kiesz M (2018) Effect of dietary supplementation with Yarrowia lipolytica or Saccharomyces cerevisiae yeast and probiotic additives on haematological parameters and the gut microbiota in piglets. Res Vet Sci 119:221–227
Czerucka D, Piche T, Rampal P (2007) Review article: yeast as probiotics- Saccharomyces boulardii. Aliment Pharmacol Ther 26(6):767–778
Ellis AE (2001) Innate host defense mechanisms of fish against viruses and bacteria. Dev Comp Immunol 25(8–9):827–839
Frigo DE, Tang Y, Beckman BS, Scandurro AB, Alam J, Burow ME, McLachlan JA (2004) Mechanism of AP-1-mediated gene expression by select organochlorines through the p38 MAPK pathway. Carcinogenesis 25(2):249–261
Gan Y, Cui X, Ma T, Liu Y, Li A, Huang M (2014) Paeoniflorin upregulates β-defensin-2 expression in human bronchial epithelial cell through the p38 MAPK, ERK, and NF-κB signaling pathways. Inflammation 37(5):1468–1475
Garcia Diaz T, Ferriani Branco A, Jacovaci FA, Cabreira Jobim C, Bolson DC, Pratti Daniel JL (2018) Inclusion of live yeast and mannan-oligosaccharides in high grain-based diets for sheep: ruminal parameters, inflammatory response and rumen morphology. PLoS One 13(2):e0193313
Haarmann H, Steiner T, Schreiber F, Heinrich A, Zweigner J, N'Guessan PD, Slevogt H (2015) The role and regulation of Moraxella catarrhalis-induced human beta- defensin 3 expression in human pulmonary epithelial cells. Biochem Biophys Res Commun 467(1):46–52
Han SH, Kim YE, Park JA, Park JB, Kim YS, Lee Y, Choi IG, Kwon HJ (2009) Expression of human beta-defensin-2 gene induced by CpG-DNA in human B cells. Biochem Biophys Res Commun 389(3):443–448
He M, Ichinose T, Song Y, Yoshida Y, Kobayashi F, Maki T, Yoshida S, Takano H, Shibamoto T, Sun G (2015) The role of toll-like receptors and myeloid differentiation factor 88 in Bjerkandera adusta-induced lung inflammation. Int Arch Allergy Immunol 168(2):96–106
He YJ, Kuchta K, Deng YM, Cameron S, Lin Y, Liu XY, Ye GR, Lv X, Kobayashi Y, Shu JC (2017) Curcumin promotes apoptosis of activated hepatic stellate cells by inhibiting protein expression of the MyD88 pathway. Planta Med 83(18):1392–1396
Hou A, Tin MQ, Tong L (2017) Toll-like receptor 2-mediated NF-kappa B pathway activation in ocular surface epithelial cells. Eye Vis 4:17
Huttner KM, Brezinski-Caliguri DJ, Mahoney MM, Diamond G (1998) Antimicrobial peptide expression is developmentally regulated in the ovine gastrointestinal tract. J Nutr 128(2 Suppl):297S–299S
Jin X, Zhang M, Zhu X, Fan Y, Du C, Bao H, Xu S, Tian Q, Wang Y, Yang Y (2018) Modulation of ovine SBD-1 expression by saccharomyces cerevisiae in ovine ruminal epithelial cells. BMC Vet Res 14(1):134
Kawai K, Shimura H, Minagawa M, Ito A, Tomiyama K, Ito M (2002) Expression of functional toll-like receptor 2 on human epidermal keratinocytes. J Dermatol Sci 30(3):185–194
Khatua S, Acharya K (2018) Water soluble Antioxidative crude polysaccharide from Russula senecis elicits TLR modulated NF-κB signaling pathway and pro-inflammatory response in murine macrophages. Front Pharmacol 9:985
Kim HA, Cho ML, Choi HY, Yoon CS, Jhun JY, Oh HJ, Kim HY (2006) The catabolic pathway mediated by toll-like receptors in human osteoarthritic chondrocytes. Arthritis Rheum 54(7):2152–2163
Kim JH, Kim KH, Kim HJ, Lee J, Myung SC (2015) Expression of Beta- Defensin 131 promotes an innate immune response in human prostate epithelial cells. PLoS One 10(12):e0144776
Lee HY, Takeshita T, Shimada J, Akopyan A, Woo JI, Pan H, Moon SK, Andalibi A, Park RK, Kang SH, Kang SS, Gellibolian R, Lim DJ (2008) Induction of beta defensin 2 by NTHi requires TLR-2 mediated MyD88 and IRAK-TRAF6-p38 MAPK signaling pathway in human middle ear epithelial cells. BMC Infect Dis 8:87
Lewis SB, Prior A, Ellis SJ, Cook V, Chan SS, Gelson W, Schüller S (2016) Flagellin induces beta-Defensin 2 in human colonic ex vivo infection with Enterohemorrhagic Escherichia coli. Front Cell Infect Microbiol 6:68
Li Y, Yang YF (2013) Research progress of polymorphism and expression of defensins in vivo of livestock. China Anim Husb Vet Med 40(3):160–168
Li Q, Bao F, Zhi D, Liu M, Yan Q, Zheng X, Ren L, Cong S, Li Y, Cao G (2016) Lipopolysaccharide induces SBD-1 expression via the P38 MAPK signaling pathway in ovine oviduct epithelial cells. Lipids Health Dis 15(1):127
Machado LR, Ottolini B (2015) An evolutionary history of defensins: a role for copy number variation in maximizing host innate and adaptive immune responses. Front Immunol 6:115
Muire PJ, Hanson LA, Wills R, Petrie-Hanson L (2017) Differential gene expression following TLR stimulation in rag1- / - mutant zebrafish issues and morphological descriptions of lymphocyte-like cell populations. PLoS One 12(9):e0184077
Niyonsaba F, Ushio H, Nagaoka I, Okumura K, Ogawa H (2005) The human beta-defensins (-1, -2, -3, -4) and cathelicidin LL-37 induce IL-18 secretion through p38 and ERK MAPK activation in primary human keratinocytes. J Immunol 175:1776–1784
Qi C, Cai Y, Gunn L, Ding C, Li B, Kloecker G, Qian K, Vasilakos J, Saijo S, Iwakura Y, Yannelli JR, Yan J (2011) Differential pathways regulating innate and adaptive antitumor immune responses by particulate and soluble yeast-derived β-glucans. Blood 117(25):6825–6836
Reyes-becerril M, Guardiola FA, Sanchez V, Maldonado M, Angulo C (2018) Sterigmatomyces halophilus β-glucan improves the immune response and bacterial resistance in pacific red snapper (lutjanus peru) peripheral blood leucocytes: in vitro study. Fish Shellfish Immunol 78:392–403
Roux PP, Blenis J (2004) ERK and p38 MAPK-activated protein kinases: a family of protein kinases with diverse biological functions. Microbiol Mol Biol Rev 68(2):320–344
Sato S, Sanjo H, Takeda K, Ninomiya-Tsuji J, Yamamoto M, Kawai T, Matsumoto K, Takeuchi O, Akira S (2005) Essential function for the kinase TAK1 in innate and adaptive immune responses. Nat Immunol 6(11):1087–1095
Takeuchi O, Akira S (2010) Pattern recognition receptors and inflammation. Cell 140(6):805–820
Wells JM, Rossi O, Meijerink M, van Baarlen P (2011) Epithelial crosstalk at the microbiota-mucosal interface. Proc Natl Acad Sci U S A 108(Suppl 1):4607–4614
Wen S, Cao G, Bao T, Cheng L, Li H, Du C, Tu Y, Li Q, Jian R, Zhao P, Wuriliga (2012) Modulation of ovine SBD-1 expression by 17beta-estradiol in ovine oviduct epithelial cells. BMC Vet Res 8:143
Winer J, Jung CK, Shackel I, Williams PM (1999) Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Anal Biochem 270(1):41–49
Wu H, Zhao G, Jiang K, Chen X, Rui G, Qiu C, Guo M, Deng G (2016) IFN-τ alleviates lipopolysaccharide-induced inflammation by suppressing NF-κB and MAPKs pathway activation in mice. Inflammation 39(3):1141–1150
Xing Z, Cardona CJ, Anunciacion J, Adams S, Dao N (2010) Roles of the ERK MAPK in the regulation of proinflammatory and apoptotic responses in chicken macrophages infected with H9N2 avian influenza virus. J Gen Virol 91(Pt 2):343–351
Yamamoto M, Takeda K (2010) Current views of toll-like receptor signaling pathways. Gastroenterol Res Pract 2010:240365
Yu Y, Nagai S, Wu H, Neish AS, Koyasu S, Gewirtz AT (2006) TLR5-mediated phosphoinositide 3-kinase activation negatively regulates flagellin-induced proinflammatory gene expression. J Immunol 176(10):6194–6201
Zhang M, Jin X, Wang YH, Wei F, Wen JY, Li ZY, Yang YF (2018) Effects of Saccharomyces cerevisiae β-glucan on the expression of SBD-1 in ovine ruminal epithelial cells. Acta Vet Zootech Sin 49(11):2416–2424
Zhao L, Lu W (2014) Defensins in innate immunity. Curr Opin Hematol 21(1):37–42
Acknowledgments
We thank Editage for their assistance in editing our article and for language revision. We thank Prof. Gui-fang Cao for providing us with laboratory space for the experiments.
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The dataset supporting the conclusions of this article is included within the article and in Additional files.
Funding
This study was supported by the National Natural Science Foundation of China (Grant No. 31560682), and they provided suggestions about the study design.
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Contributions
XJ designed the experiments, cultured the cells, extracted RNA and prepared cDNA, performed western blotting, the statistical analysis, prepared the manuscript and figures; MZ performed qPCR and ELISA experiments, statistical analysis, and prepared the manuscript and figures; YY designed the experiments and revised the manuscript. All authors have read and approved the final manuscript.
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All animal treatments and procedures used in the present study were approved by the Institutional Animal Care and Use Committee of the IMAU with adherence to IMAU guidelines.
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Implications
β-Glucan, a cell wall component of Saccharomyces cerevisiae, can stimulate innate immune responses and trigger the up-regulation of SBD-1 in ORECs. This fundamental study could provide a reference on the regulation mechanism of TLR-2-MyD88-NF-κB /MAPK signaling on β-glucan-stimulated rumen and would contribute to the understanding of immunological modulations in the gastrointestinal track triggered by probiotic yeast cell wall components. This study would also shed light on gastrointestinal tract-microbiota-mediated immune reactions.
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Jin, X., Zhang, M. & Yang, Yf. Saccharomyces cerevisiae β-glucan-induced SBD-1 expression in ovine ruminal epithelial cells is mediated through the TLR-2-MyD88-NF-κB/MAPK pathway. Vet Res Commun 43, 77–89 (2019). https://doi.org/10.1007/s11259-019-09747-x
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DOI: https://doi.org/10.1007/s11259-019-09747-x