Effect of Promoter Methylation on the Expression of Porcine MUC2 Gene and Resistance to PEDV Infection

Integrity of the intestinal mucosal barrier is closely related to the occurrence of diarrhea. As an important component protein of the intestinal mucosal barrier, Mucin 2 (MUC2) plays a critical role in preventing the invasion of pathogens, toxins, and foreign bodies. In the present study, we preliminary verified the function of the porcine MUC2 gene in resisting porcine epidemic diarrhea virus (PEDV) infection and investigated the effect of DNA methylation in the promoter region on MUC2 gene expression. The results showed that after PEDV infection, the intestinal mucosal barrier was damaged. Moreover, MUC2 expression was significantly higher in PEDV-infected piglets than in healthy piglets (P < 0.01). The mRNA expression of MUC2 was significantly higher in PEDV-infected IPEC-J2 cells than in non-infected IPEC-J2 cells (P < 0.05). Methylation of the mC-5 site in the MUC2 promoter inhibited the binding of Yin Yang 1 (YY1) to the promoter, down regulated the expression of MUC2 and increased the susceptibility of piglets to PEDV. In conclusion, this study suggests that MUC2 plays an essential regulatory role in PEDV infection. High MUC2 expression improves the resistance of pigs to PEDV infection. The binding of YY1 to the MUC2 promoter is hindered by the methylation of the mC-5 site, which downregulates MUC2 expression and ultimately affects the resistance of pigs to PEDV infection.

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Analysis of Intestinal Mucosa Integrity and GLP-2 Gene Functions upon Porcine Epidemic Diarrhea Virus Infection in Pigs

Animals ◽

10.3390/ani11030644 ◽

2021 ◽

Vol 11 (3) ◽

pp. 644

Author(s):

Yajing Zhou ◽

Zhanshi Ren ◽

Shuai Zhang ◽

Haifei Wang ◽

Shenglong Wu ◽

...

Keyword(s):

Epithelial Cells ◽

Intestinal Mucosa ◽

Porcine Epidemic Diarrhea Virus ◽

Intestinal Epithelial Cells ◽

Mrna Level ◽

Mucosal Barrier ◽

Intestinal Epithelial ◽

Intestinal Mucosal Barrier ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

Porcine epidemic diarrhea virus (PEDV) infects intestinal epithelial cells, destroys the intestinal mucosal barrier and then causes diarrhea in piglets. Glucagon-like peptide-2 (GLP-2) is a specific intestinal growth hormone that promotes the repair of damaged intestinal mucosa and improves the intestinal barrier. In this study, we investigated the functions of porcine GLP-2 gene in regulating PEDV infection. The intestinal tissues with damaged intestinal structures caused by PEDV infection were first confirmed and collected. Expression analysis indicated that the GLP-2 gene was expressed in the duodenum, jejunum and ileum tissues, and the mRNA level was significantly down-regulated in jejunum and ileum of piglets with damaged intestinal mucosa. Infection of PEDV to porcine small intestinal epithelial cells in vitro showed that GLP-2 gene was significantly decreased, which was consistent with the expression pattern in intestinal tissues. In addition, we silenced the GLP-2 gene by shRNA interfering and found that the copy numbers of PEDV were remarkably increased in the GLP-2 gene silencing cells. Our findings suggest that the GLP-2 gene was potentially involved in regulating PEDV infection and in maintaining the integrity of the intestinal mucosal barrier structure, which could contribute to our understanding of the mechanisms of PEDV pathogenesis and provide a theoretical basis for the identification and application of resistant genes in pig selective breeding for porcine epidemic diarrhea.

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Porcine amino peptidase N domain VII has critical role in binding and entry of porcine epidemic diarrhea virus

Virus Research ◽

10.1016/j.virusres.2016.10.004 ◽

2017 ◽

Vol 227 ◽

pp. 150-157 ◽

Cited By ~ 8

Author(s):

Anthony Ndirangu Kamau ◽

Jung-Eun Park ◽

Eui-Soon Park ◽

Jung-Eun Yu ◽

Jaerang Rho ◽

...

Keyword(s):

Porcine Epidemic Diarrhea Virus ◽

Critical Role ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

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Manipulation of Intestinal Antiviral Innate Immunity and Immune Evasion Strategies of Porcine Epidemic Diarrhea Virus

BioMed Research International ◽

10.1155/2019/1862531 ◽

2019 ◽

Vol 2019 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Jian Du ◽

Junqiu Luo ◽

Jie Yu ◽

Xiangbing Mao ◽

Yuheng Luo ◽

...

Keyword(s):

Innate Immunity ◽

Immune Evasion ◽

Porcine Epidemic Diarrhea Virus ◽

Current Knowledge ◽

Mucosal Barrier ◽

Watery Diarrhea ◽

Diarrhea Virus ◽

Successful Infection ◽

Porcine Epidemic Diarrhea ◽

Antiviral Innate Immunity

Porcine epidemic diarrhea virus (PEDV) infection causes watery diarrhea, dehydration, and high mortality in neonatal pigs, due to its clinical pathogenesis of the intestinal mucosal barrier dysfunction. The host’s innate immune system is the first line of defence upon virus invasion of the small intestinal epithelial cells. In turn, the virus has evolved to modulate the host’s innate immunity during infection, resulting in pathogen virulence, survival, and the establishment of successful infection. In this review, we gather current knowledge concerning the interplay between PEDV and components of host innate immunity, focusing on the role of cytokines and interferons in intestinal antiviral innate immunity, and the mechanisms underlying the immune evasion strategies of PEDV invasion. Finally, we provide some perspectives on the potential prevention and treatment for PEDV infection.

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Diverse expression patterns of mucin 2 in colorectal cancer indicates its mechanism related to the intestinal mucosal barrier

World Journal of Gastroenterology ◽

10.3748/wjg.v27.i25.3888 ◽

2021 ◽

Vol 27 (25) ◽

pp. 3888-3900

Author(s):

Guo-Lian Gan ◽

Hua-Tao Wu ◽

Wen-Jia Chen ◽

Chun-Lan Li ◽

Qian-Qian Ye ◽

...

Keyword(s):

Colorectal Cancer ◽

Expression Patterns ◽

Mucosal Barrier ◽

Intestinal Mucosal Barrier ◽

Mucin 2

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Lack of LTβR Increases Susceptibility of IPEC-J2 Cells to Porcine Epidemic Diarrhea Virus

Cells ◽

10.3390/cells7110222 ◽

2018 ◽

Vol 7 (11) ◽

pp. 222 ◽

Cited By ~ 3

Author(s):

Tawfeek Altawaty ◽

Lulu Liu ◽

Hongyong Zhang ◽

Cong Tao ◽

Shaohua Hou ◽

...

Keyword(s):

Cell Proliferation ◽

Porcine Epidemic Diarrhea Virus ◽

Cellular Response ◽

Target Genes ◽

Cell Types ◽

Digital Holographic Microscopy ◽

Mucosal Barrier ◽

Label Free ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

The essential requirement of the lymphotoxin beta receptor (LTβR) in the development and maintenance of peripheral lymphoid organs is well recognized. Evidence shows that LTβR is involved in various cellular processes; however, whether it plays a role in maintaining the cellular function of intestinal porcine enterocytes (IPEC-J2), specifically during porcine epidemic diarrhea virus (PEDV) infection, remains unknown. In this study, we generated LTβR null IPEC-J2 cells using CRISPR/Cas9 to examine the importance of LTβR in cell proliferation, apoptosis, and the response to PEDV infection. Our results showed that the lack of LTβR leads to significantly decreased cell proliferation, potentially due to S phase arrest in LTβR−/− IPEC-J2 cells. Label-free digital holographic microscopy was used to record the three-dimensional morphology of both cell types for up to 72 hours and revealed significantly increased numbers of LTβR−/− cells undergoing apoptosis. Furthermore, we found that PEDV-infected LTβR−/− null IPEC-J2 cells exhibited significant suppression of nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) target genes (interleukin (IL)-6 and IL-8) and mucosal barrier integrity-related genes (vascular cell adhesion molecule 1 (VCAM1) and IL-22), which may explain why LTβR−/− cells are more susceptible to PEDV infection. Collectively, our data not only demonstrate the key role of LTβR in intestinal porcine enterocytes, but also provide data for the improved understanding of the cellular response to PEDV infection.

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Regulatory Effect of Methylation of the Porcine AQP3 Gene Promoter Region on Its Expression Level and Porcine Epidemic Diarrhea Virus Resistance

Genes ◽

10.3390/genes11101167 ◽

2020 ◽

Vol 11 (10) ◽

pp. 1167

Author(s):

Jia-Yun Wu ◽

Fang Wang ◽

Zheng-Chang Wu ◽

Sheng-Long Wu ◽

Wen-Bin Bao

Keyword(s):

Porcine Epidemic Diarrhea Virus ◽

Promoter Region ◽

Gene Expression Regulation ◽

Cpg Islands ◽

Intestinal Barrier ◽

Pcr Amplification ◽

Intestinal Secretion ◽

Significant Negative Correlation ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

As an important carrier for intestinal secretion and water absorption, aquaporin 3 (AQP3) is closely related to diarrhea. In this study, we investigated the mechanisms of AQP3 gene expression regulation in porcine epidemic diarrhea virus (PEDV)-induced diarrhea confirmed by PCR amplification and sequencing. Evaluation of intestinal pathology showed that diarrhea caused by PEDV infection destroyed the intestinal barrier of piglets. qPCR analysis showed that AQP3 expression in the small intestine of PEDV-infected piglets was extremely significantly decreased. qPCR and Bisulfite sequencing PCR revealed an increase in the methylation levels of both CpG islands in the AQP3 promoter region in the jejunum of PEDV-infected piglets. The methylation of mC-20 and mC-10 sites within the two CpG islands showed a significant negative correlation with AQP3 expression. Chromatin Co-Immunoprecipitation (ChIP)-PCR showed that the Sp1 transcription factor was bound to the AQP3 promoter region containing these two CpG sites. AQP3 expression was also extremely significantly reduced in Sp1-inhibited IPEC-J2 cells, indicating that abnormal methylation at the mC-20 site of CpG1 and the mC-10 site of CpG2 reduces its expression in PEDV-infected piglet jejunum by inhibiting the binding of Sp1 to the AQP3 promoter. These findings provide a theoretical basis for further functional studies of porcine AQP3.

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