Immortalized porcine intestinal epithelial cell cultures susceptible to porcine rotavirus infection

Wang, Jing; Hu, Guang‐Wan; Gao, Wanjun; Xu, Lei; Ning, Pengbo; Zhang, Yanming

doi:10.1016/j.jviromet.2014.03.007

Cited by 7 publications

(7 citation statements)

References 32 publications

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“…EC (swine umbilical vein endothelial cell) [47] and IEC (swine intestinal epithelial cell) [48] were immortalized porcine cell lines established and stored by our lab (Yanming, Zhang lab). PK (or PK-15, porcine kidney epithelial) [49] and ST [50] Cells for all assays relating to CSFV infection were cultured to a confluent of 60% in 35 mm cell-culture dishes in MEM containing 10% fetal serum and then incubated with 1.5 ml CSFV liquid (100 CCID50/0.1 ml) at 37uC, 5% CO 2 for 1 h. Subsequently, supernatant was discarded and the CSFV incubated cells were washed with fresh MEM to remove the unattached virions before the further cultivation in the final culture medium of MEM with 2% fetal serum at 37uC, 5% CO 2 .…”

Section: Cell Lines Reagents and Csfv Infectionmentioning

confidence: 99%

Integrin β3 Is Required in Infection and Proliferation of Classical Swine Fever Virus

Wang

Liang

et al. 2014

PLoS ONE

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Classical Swine Fever (CSF) is a highly infectious fatal pig disease, resulting in huge economic loss to the swine industry. Integrins are membrane-bound signal mediators, expressed on a variety of cell surfaces and are known as receptors or co-receptors for many viruses. However, the role of integrin β3 in CSFV infection is unknown. Here, through quantitive PCR, immunofluorescence (IFC) and immunocytohistochemistry (ICC), we revealed that ST (swine testicles epithelial) cells have a prominent advantage in CSFV proliferation as compared to EC (swine umbilical vein endothelial cell), IEC (swine intestinal epithelial cell) and PK (porcine kidney epithelial) cells. Meanwhile, ST cells had remarkably more integrin β3 expression as compared to EC, IEC and PK cells, which was positively correlated with CSFV infection and proliferation. Integrin β3 was up-regulated post CSFV infection in all the four cell lines, while the CSFV proliferation rate was decreased in integrin β3 function-blocked cells. ShRNA1755 dramatically decreased integrin β3, with a deficiency of 96% at the mRNA level and 80% at the protein level. CSFV proliferation was dramatically reduced in integrin β3 constantly-defected cells (ICDC), with the deficiencies of 92.6%, 99% and 81.7% at 24 h, 48 h and 72 h post CSFV infection, respectively. These results demonstrate that integrin β3 is required in CSFV infection and proliferation, which provide a new insight into the mechanism of CSFV infection.

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Section: Cell Lines Reagents and Csfv Infectionmentioning

confidence: 99%

Integrin β3 Is Required in Infection and Proliferation of Classical Swine Fever Virus

Wang

Liang

et al. 2014

PLoS ONE

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“…Studies have used human colon adenocarcinoma (Caco-2) and carcinoma (HT-29) cell lines, and Madin-Darby canine kidney (MDCK) and rhesus monkey kidney (MA104) cell lines to study RVs infection or host-pathogen interactions in vitro (reviewed in [ 10 ]). Of interest, Caco-2 and HT-29 cells are tumorigenic lines and it was found that they possess different phenotypes compared with normal cells therefore; they would not be able to mimic exactly the behavior of IECs in response to the challenge with RVs [ 11 ]. The porcine small intestinal epithelial cell line (IPEC-J2) has been proposed as model for the study of innate immune responses to RVs.…”

Section: Introductionmentioning

confidence: 99%

Immunobiotic Bifidobacteria Strains Modulate Rotavirus Immune Response in Porcine Intestinal Epitheliocytes via Pattern Recognition Receptor Signaling

et al. 2016

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In this work, we aimed to characterize the antiviral response of an originally established porcine intestinal epithelial cell line (PIE cells) by evaluating the molecular innate immune response to rotavirus (RVs). In addition, we aimed to select immunomodulatory bacteria with antiviral capabilities. PIE cells were inoculated with RVs isolated from different host species and the infective titers and the molecular innate immune response were evaluated. In addition, the protection against RVs infection and the modulation of immune response by different lactic acid bacteria (LAB) strains was studied. The RVs strains OSU (porcine) and UK (bovine) effectively infected PIE cells. Our results also showed that RVs infection in PIE cells triggered TLR3-, RIG-I- and MDA-5-mediated immune responses with activation of IRF3 and NF-κB, induction of IFN-β and up-regulation of the interferon stimulated genes MxA and RNase L. Among the LAB strains tested, Bifidobacterium infantis MCC12 and B. breve MCC1274 significantly reduced RVs titers in infected PIE cells. The beneficial effects of both bifidobacteria were associated with reduction of A20 expression, and improvements of IRF-3 activation, IFN-β production, and MxA and RNase L expressions. These results indicate the value of PIE cells for studying RVs molecular innate immune response in pigs and for the selection of beneficial bacteria with antiviral capabilities.

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“…To explore whether PEDV infection upregulates miR-221-5p expression, we infected MARC-145 cells with the PEDV strain CH/HBTS/2017 at a multiplicity of infection (MOI) of 1. After 24 h, real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) results confirmed that miR-221-5p expression was significantly upregulated in PEDV-infected MARC-145 cells ( Figure 1 A), with this finding verified in PEDV CV777-infected porcine intestinal epithelial cells (IECs) [ 21 ]. The 50% tissue culture infectious dose (TCID 50 ) and immunofluorescence analysis were performed to confirm virus infection ( Figure 1 B,C).…”

Section: Resultsmentioning

confidence: 81%

“…In the present study, we initially evaluated whether miR-221-5p was upregulated by PEDV infection using MARC-145 cells, which are suitable for studying PEDV infection and innate immune modulation [ 33 ]. Additionally, we used PEDV strains CH/HBTS/2017 and CV777, as well as porcine IECs harboring epithelial cell markers, and incapable of withstanding the higher trypsin concentrations (>2.5 µg/mL) necessary for evaluating CV777 infection [ 21 ]. Infection with both PEDV strains significantly upregulated miR-221-5p expression in the host cells ( Figure 1 ), suggesting its potential role in cellular response to PEDV infection.…”

Section: Discussionmentioning

confidence: 99%

MicroRNA-221-5p Inhibits Porcine Epidemic Diarrhea Virus Replication by Targeting Genomic Viral RNA and Activating the NF-κB Pathway

Zheng

Liu³

et al. 2018

IJMS

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MicroRNAs (miRNAs) are a class of noncoding RNAs involved in posttranscriptional regulation of gene expression and many critical roles in numerous biological processes. Porcine epidemic diarrhea virus (PEDV), the etiological agent of porcine epidemic diarrhea, causes substantial economic loss in the swine industry worldwide. Previous studies reported miRNA involvement in viral infection; however, their role in regulating PEDV infection remains unknown. In this study, we investigated the regulatory relationship between miRNA-221-5p and PEDV infection, finding that miR-221-5p overexpression inhibited PEDV replication in a dose-dependent manner, and that silencing endogenous miR-221-5p enhanced viral replication. Our results showed that miR-221-5p directly targets the 3′ untranslated region (UTR) of PEDV genomic RNA to inhibit PEDV replication, and that miR-221-5p overexpression activates nuclear factor (NF)-κB signaling via p65 nuclear translocation, thereby upregulating interferon (IFN)-β, IFN-stimulated gene 15, and MX1 expression during CH/HBTS/2017 infection. We subsequently identified NF-κB-inhibitor α and suppressor of cytokine signaling 1, negative regulators of the NF-κB pathway, as miR-221-5p targets. These results demonstrated the ability of miR-221-5p to inhibit PEDV replication by targeting the 3’ UTR of the viral genome and activating the NF-κB-signaling pathway. Our findings will aid the development of preventive and therapeutic strategies for PEDV infection.

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Immortalized porcine intestinal epithelial cell cultures susceptible to porcine rotavirus infection

Cited by 7 publications

References 32 publications

Integrin β3 Is Required in Infection and Proliferation of Classical Swine Fever Virus

Integrin β3 Is Required in Infection and Proliferation of Classical Swine Fever Virus

Immunobiotic Bifidobacteria Strains Modulate Rotavirus Immune Response in Porcine Intestinal Epitheliocytes via Pattern Recognition Receptor Signaling

MicroRNA-221-5p Inhibits Porcine Epidemic Diarrhea Virus Replication by Targeting Genomic Viral RNA and Activating the NF-κB Pathway

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