Document Type : Original Article
Authors
- Yan Jiang
- Linqiao Li
- Qilu Pan
- Xiaojing Du
- Qian Han
- Feixiang Ling
- Rou Li
- Lin Mai
- Jianwei Huang
- Shuyuan Chu
- Libing Ma
Department of Respiratory and Critical Care Medicine, Affiliated Hospital of Guilin Medical University, Guilin 541001, Guangxi, China.
Abstract
Background: Little is known about MBD2’s epigenetic regulation in the immune pathogenesis of CD4+T cell differentiation.
Objective: This study attempted to explore the mechanism of methyl-cpg-binding domain protein 2 (MBD2) in CD4+T cell differentiation stimulated by environmental allergen ovalbumin (OVA).
Methods: Mononuclear cells were separated from the spleen tissues of male C57BL/6 mice. The OVA interfered with the differentiation of splenic mononuclear cells and CD4+T cells. The CD4+T cells were obtained by magnetic beads and identified by CD4 labeled antibody. CD4+T cells were transfected with lentivirus to silence MBD2 gene. A methylation quantification kit was used to detect 5-mC levels.
Results: The purity of CD4+T cells reached 95.99% after magnetic beads sorting. Treatment with 200 μg/mL OVA stimulated the CD4+T cells differentiation to Th17 cells and promoted the secretion of IL-17. After being induced, the Th17 cell ratio increased. 5-Aza inhibited the Th17 cell differentiation and the IL-17 level in a dose-dependent manner. Under the intervention of the Th17 induction and 5-Aza, MBD2 silencing inhibited the differentiation of Th17 cell, and decreased the IL-17 and 5-mC levels in the cell supernatants. MBD2 silencing reduced the scale of the Th17 cell and IL-17 levels in the OVA-treated CD4+T cells.
Conclusion: MBD2 affected IL-17 and 5-mC levels by mediating the Th17 cell differentiation in splenic CD4+T cells that were interfered with 5-Aza. OVA induced Th17 differentiation and increased IL-17 levels, inhibited by MBD2 silencing.
Keywords
- Stern J, Pier J, Litonjua AA. Asthma epidemiology and risk factors. Semin Immunopathol. 2020; 42:5-15.
- Alwarith J, Kahleova H, Crosby L, et al. The role of nutrition in asthma prevention and treatment. Nutr Rev. 2020; 78:928-38.
- Fernandes JS, Araujo MI, de Almeida T, et al. Impaired immunoregulatory network of the CD4 T lymphocytes in refractory asthma. Clin Exp Allergy. 2019; 49:644-54.
- Chen X, Lin H, Yang D, et al. Early-life undernutrition reprograms CD4(+) T-cell glycolysis and epigenetics to facilitate asthma. J Allergy Clin Immunol. 2019; 143:2038-51.e12.
- Ling MF, Luster AD. Allergen-Specific CD4(+) T Cells in Human Asthma. Ann Am Thorac Soc. 2016; 13 Suppl 1:S25-30.
- Seumois G, Ramírez-Suástegui C, Schmiedel BJ, et al. Single-cell transcriptomic analysis of allergen-specific T cells in allergy and asthma. Sci Immunol. 2020; 5:eaba6087.
- Luckheeram RV, Zhou R, Verma AD, Xia B. CD4⁺T cells: differentiation and functions. Clin Dev Immunol. 2012; 2012:925135.
- Pei YF, Xu XN, Wang ZF, et al. Methyl-CpG Binding Domain Protein 2 Inhibits the Malignant Characteristic of Lung Adenocarcinoma through the Epigenetic Modulation of 10 to 11 Translocation 1 and miR-200s. Am J Pathol. 2019; 189:1065-76.
- McGrath-Morrow SA, Ndeh R, Helmin KA, et al. DNA methylation regulates the neonatal CD4(+) T-cell response to pneumonia in mice. J Biol Chem. 2018; 293:11772-83.
- Du Q, Luu PL, Stirzaker C, Clark SJ. Methyl-CpG-binding domain proteins: readers of the epigenome. Epigenomics. 2015; 7:1051-73.
- Sato M, Horio Y, Sekido Y, Minna JD, Shimokata K, Hasegawa Y. The expression of DNA methyltransferases and methyl-CpG-binding proteins is not associated with the methylation status of p14(ARF), p16(INK4a) and RASSF1A in human lung cancer cell lines. Oncogene. 2002; 21:4822-9.
- McCabe DC, Caudill MA. DNA methylation, genomic silencing, and links to nutrition and cancer. Nutr Rev. 2005; 63:183-95.
- Wood KH, Zhou Z. Emerging Molecular and Biological Functions of MBD2, a Reader of DNA Methylation. Front Genet. 2016; 7:93.
- Wang Y, Zhang L, Wu GR, et al. MBD2 serves as a viable target against pulmonary fibrosis by inhibiting macrophage M2 program. Sci Adv. 2021; 7:eabb6075.
- Liu D, Tan Y, Bajinka O, Wang L, Tang Z. Th17/IL-17 Axis Regulated by Airway Microbes Get Involved in the Development of Asthma. Curr Allergy Asthma Rep. 2020; 20:11.
- Samary Cdos S, Antunes MA, Silva JD, et al. Impact of Bacillus Calmette-Guérin Moreau vaccine on lung remodeling in experimental asthma. Respir Physiol Neurobiol. 2013; 189:614-23.
- Dong M, Wang WQ, Chen J, et al. Acupuncture Regulates the Balance of CD4(+) T Cell Subtypes in Experimental Asthma Mice. Chin J Integr Med. 2019; 25:617-24.
- Jia A, Wang Y, Sun W, et al. MBD2 Regulates Th17 Cell Differentiation and Experimental Severe Asthma by Affecting IRF4 Expression. Mediators Inflamm. 2017; 2017:6249685.
- Xu L, Sun WJ, Jia AJ, et al. MBD2 regulates differentiation and function of Th17 cells in neutrophils- dominant asthma via HIF-1α. J Inflamm (Lond). 2018; 15:15.
- Sun W, Xiao B, Jia A, et al. MBD2-mediated Th17 differentiation in severe asthma is associated with impaired SOCS3 expression. Exp Cell Res. 2018; 371:196-204.
- Sun W, Yuan Y, Qiu L, et al. Increased Proportion of Dual-Positive Th2-Th17 Cells Promotes a More Severe Subtype of Asthma. Can Respir J. 2021; 2021:9999122.
- Lu Y, Kared H, Tan SW, et al. Dynamics of helper CD4 T cells during acute and stable allergic asthma. Mucosal Immunol. 2018; 11:1640-52.
- Zissler UM, Esser-von Bieren J, Jakwerth CA, Chaker AM, Schmidt-Weber CB. Current and future biomarkers in allergic asthma. Allergy. 2016; 71:475-94.
- Flaherty S, Reynolds JM. Mouse Naïve CD4+ T Cell Isolation and In vitro Differentiation into T Cell Subsets. J Vis Exp. 2015; 16:52739.
- Varanasi SK, Reddy PBJ, Bhela S, Jaggi U, Gimenez F, Rouse BT. Azacytidine Treatment Inhibits the Progression of Herpes Stromal Keratitis by Enhancing Regulatory T- Cell Function. J Virol. 2017; 91:e02367-16.
- Chan MW, Chang CB, Tung CH, Sun J, Suen JL, Wu SF. Low-dose 5-aza-2'-deoxycytidine pretreatment inhibits experimental autoimmune encephalomyelitis by induction of regulatory T cells. Mol Med. 2014; 20:248-56.
- Sun L, Fu J, Lin SH, et al. Particulate matter of 2.5 μm or less in diameter disturbs the balance of T(H)17/regulatory T cells by targeting glutamate oxaloacetate transaminase 1 and hypoxia-inducible factor 1α in an asthma model. J Allergy Clin Immunol. 2020; 145:402-14.
- Cook PC, Owen H, Deaton AM, et al. A dominant role for the methyl-CpG-binding protein Mbd2 in controlling Th2 induction by dendritic cells. Nat Commun. 2015; 6:6920.
- Santangelo S, Cousins DJ, Winkelmann NE, Staynov DZ. DNA methylation changes at human Th2 cytokine genes coincide with DNase I hypersensitive site formation during CD4(+) T cell differentiation. J Immunol. 2002; 169:1893-903.
- Zhong J, Yu Q, Yang P, et al. MBD2 regulates TH17 differentiation and experimental autoimmune encephalomyelitis by controlling the homeostasis of T-bet/Hlx axis. J Autoimmun. 2014; 53:95-104.
- Irvin C, Zafar I, Good J, et al. Increased frequency of dual-positive TH2/TH17 cells in bronchoalveolar lavage fluid characterizes a population of patients with severe asthma. J Allergy Clin Immunol. 2014; 134:1175-86.e7.
- Liu W, Liu S, Verma M, et al. Mechanism of T(H)2/T(H)17-predominant and neutrophilic T(H)2/T(H)17-low subtypes of asthma. J Allergy Clin Immunol. 2017; 139:1548-58.e4.
- Hongyan L. Esculetin Attenuates Th2 and Th17 Responses in an Ovalbumin-Induced Asthmatic Mouse Model. Inflammation. 2016; 39:735-43.
- Zhao S, Jiang Y, Yang X, et al. Lipopolysaccharides promote a shift from Th2-derived airway eosinophilic inflammation to Th17-derived neutrophilic inflammation in an ovalbumin-sensitized murine asthma model. J Asthma. 2017; 54:447-55.
- May S, Owen H, Phesse TJ, et al. Mbd2 enables tumourigenesis within the intestine while preventing tumour-promoting inflammation. J Pathol. 2018; 245:270-82.
- Wang L, Liu Y, Han R, et al. Mbd2 promotes foxp3 demethylation and T-regulatory-cell function. Mol Cell Biol. 2013; 33:4106-15.
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