The Characterization of Immunoprotection Induced by a cDNA Clone Derived from the Attenuated Taiwan Porcine Epidemic Diarrhea Virus Pintung 52 Strain

The porcine epidemic diarrhea virus (PEDV) poses a great threat to the global swine industries and the unreliable protection induced by the currently available vaccines remains a major challenge. We previously generated a genogroup 2b (G2b) PEDV Taiwan Pintung 52 (PEDVPT) strain, PEDVPT-P96, and determined its promising host immune response against the virulent PEDVPT-P5 strain. To study the attenuation determinants of PEDVPT-P96 and establish a PEDVPT-P96-based recombinant vector as a vaccine platform for further antigenicity modification, iPEDVPT-P96, a full-length cDNA clone of PEDVPT-P96, was established. Comparing to the parental PEDVPT-P96 virus, the iPEDVPT-P96 virus showed efficient replication kinetics with a delayed decline of viral load and similar but much more uniform plaque sizes in Vero cells. In the 5-week-old piglet model, fecal viral shedding was observed in the PEDVPT-P96-inoculated piglets, whereas those inoculated with iPEDVPT-P96 showed neither detectable fecal viral shedding nor PEDV-associated clinical signs. Moreover, inoculation with iPEDVPT-P96 elicited comparable levels of anti-PEDV specific plasma IgG and fecal/salivary IgA, neutralizing antibody titers, and similar but less effective immunoprotection against the virulent PEDVPT-P5 challenge compared to the parental PEDVPT-P96. In the present study, an infectious cDNA clone of an attenuated G2b PEDV strain was successfully generated for the first time, and the in vitro and in vivo data indicate that iPEDVPT-P96 is further attenuated but remains immunogenic compared to its parental PEDVPT-P96 viral stock. The successful development of the iPEDVPT-P96 cDNA clone could allow for the manipulation of the viral genome to study viral pathogenesis and facilitate the rapid development of effective vaccines.

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Systemic and Oral Immunogenicity of Porcine Epidemic Diarrhea Virus Antigen Fused to Poly-Fc of Immunoglobulin G and Expressed in ΔXT/FT Nicotiana benthamiana Plants

Frontiers in Pharmacology ◽

10.3389/fphar.2021.653064 ◽

2021 ◽

Vol 12 ◽

Author(s):

Nguyen-Quang-Duc Tien ◽

Moon-Sik Yang ◽

Yong-Suk Jang ◽

Tae-Ho Kwon ◽

Rajko Reljic ◽

...

Keyword(s):

Immunoglobulin G ◽

Porcine Epidemic Diarrhea Virus ◽

Virus Antigen ◽

Vaccine Platform ◽

Diarrhea Virus ◽

Mucosal Vaccination ◽

Porcine Epidemic Diarrhea ◽

Cellular Immune ◽

Farming Industry

Porcine epidemic diarrhea virus (PEDV), a member of the Coronaviridae family has become increasingly probelmatic in the pig farming industry. Currently, there are no effective, globally applicable vaccines against PEDV. Here, we tested a recombinant PEDV vaccine candidate based on the expression of the core neutralising epitope (COE) of PEDV conjugated to polymeric immunoglobulin G scaffold (PIGS) in glycoengineered Nicotiana benthamiana plants. The biological activity of COE-PIGS was demonstrated by binding to C1q component of the complement system, as well as the surface of antigen-presenting cells (APCs) in vitro. The recombinant COE-PIGS induced humoral and cellular immune responses specific for PEDV after both systemic and mucosal vaccination. Altogether, the data indicated that PEDV antigen fusion to poly-Fc could be a promising vaccine platform against respiratory PEDV infection.

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Parenterally Administered Porcine Epidemic Diarrhea Virus-Like Particle-Based Vaccine Formulated with CCL25/28 Chemokines Induces Systemic and Mucosal Immune Protectivity in Pigs

Viruses ◽

10.3390/v12101122 ◽

2020 ◽

Vol 12 (10) ◽

pp. 1122

Author(s):

Chin-Wei Hsu ◽

Ming-Hao Chang ◽

Hui-Wen Chang ◽

Tzong-Yuan Wu ◽

Yen-Chen Chang

Keyword(s):

Cellular Immunity ◽

Porcine Epidemic Diarrhea Virus ◽

Protective Efficacy ◽

Clinical Signs ◽

Effective Vaccine ◽

Control Group ◽

Diarrhea Virus ◽

Viral Shedding ◽

Specific Igg ◽

Porcine Epidemic Diarrhea

Generation of a safe, economical, and effective vaccine capable of inducing mucosal immunity is critical for the development of vaccines against enteric viral diseases. In the current study, virus-like particles (VLPs) containing the spike (S), membrane (M), and envelope (E) structural proteins of porcine epidemic diarrhea virus (PEDV) expressed by the novel polycistronic baculovirus expression vector were generated. The immunogenicity and protective efficacy of the PEDV VLPs formulated with or without mucosal adjuvants of CCL25 and CCL28 (CCL25/28) were evaluated in post-weaning pigs. While pigs intramuscularly immunized with VLPs alone were capable of eliciting systemic anti-PEDV S-specific IgG and cellular immunity, co-administration of PEDV VLPs with CCL25/28 could further modulate the immune responses by enhancing systemic anti-PEDV S-specific IgG, mucosal IgA, and cellular immunity. Upon challenge with PEDV, both VLP-immunized groups showed milder clinical signs with reduced fecal viral shedding as compared to the control group. Furthermore, pigs immunized with VLPs adjuvanted with CCL25/28 showed superior immune protection against PEDV. Our results suggest that VLPs formulated with CCL25/28 may serve as a potential PEDV vaccine candidate and the same strategy may serve as a platform for the development of other enteric viral vaccines.

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Differential expression and correlation analysis of miRNA–mRNA profiles in swine testicular cells infected with porcine epidemic diarrhea virus

Scientific Reports ◽

10.1038/s41598-021-81189-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Xiaoqian Zhang ◽

Chang Li ◽

Bingzhou Zhang ◽

Zhonghua Li ◽

Wei Zeng ◽

...

Keyword(s):

Differential Expression ◽

Porcine Epidemic Diarrhea Virus ◽

Expression Profiles ◽

Expression Patterns ◽

Bacterial Invasion ◽

Sequencing Data ◽

Diarrhea Virus ◽

Comparison Groups ◽

Porcine Epidemic Diarrhea

AbstractThe variant virulent porcine epidemic diarrhea virus (PEDV) strain (YN15) can cause severe porcine epidemic diarrhea (PED); however, the attenuated vaccine-like PEDV strain (YN144) can induce immunity in piglets. To investigate the differences in pathogenesis and epigenetic mechanisms between the two strains, differential expression and correlation analyses of the microRNA (miRNA) and mRNA in swine testicular (ST) cells infected with YN15, YN144, and mock were performed on three comparison groups (YN15 vs Control, YN144 vs Control, and YN15 vs YN144). The mRNA and miRNA expression profiles were obtained using next-generation sequencing (NGS), and the differentially expressed (DE) (p-value < 0.05) mRNA and miRNA were obtained using DESeq R package. mRNAs targeted by DE miRNAs were predicted using the miRanda algortithm. 8039, 8631 and 3310 DE mRNAs, and 36, 36, and 22 DE miRNAs were identified in the three comparison groups, respectively. 14,140, 15,367 and 3771 DE miRNA–mRNA (targeted by DE miRNAs) interaction pairs with negatively correlated expression patterns were identified, and interaction networks were constructed using Cytoscape. Six DE miRNAs and six DE mRNAs were randomly selected to verify the sequencing data by real-time relative quantitative reverse transcription polymerase chain reaction (qRT-PCR). Based on bioinformatics analysis, we discovered the differences were mostly involved in host immune responses and viral pathogenicity, including NF-κB signaling pathway and bacterial invasion of epithelial cells, etc. This is the first comprehensive comparison of DE miRNA–mRNA pairs in YN15 and YN144 infection in vitro, which could provide novel strategies for the prevention and control of PED.

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Characterization of a pathogenic full-length cDNA clone of a virulent porcine epidemic diarrhea virus strain AH2012/12 in China

Virology ◽

10.1016/j.virol.2016.10.011 ◽

2017 ◽

Vol 500 ◽

pp. 50-61 ◽

Cited By ~ 22

Author(s):

Baochao Fan ◽

Zhengyu Yu ◽

Fengjiao Pang ◽

Xiangwei Xu ◽

Baimeng Zhang ◽

...

Keyword(s):

Virus Strain ◽

Cdna Clone ◽

Porcine Epidemic Diarrhea Virus ◽

Full Length ◽

Diarrhea Virus ◽

Full Length Cdna ◽

Porcine Epidemic Diarrhea

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Short communication: antiviral activity of porcine IFN-λ3 against porcine epidemic diarrhea virus in vitro

Virus Genes ◽

10.1007/s11262-016-1374-2 ◽

2016 ◽

Vol 52 (6) ◽

pp. 877-882 ◽

Cited By ~ 9

Author(s):

Haiyan Shen ◽

Chunhong Zhang ◽

Pengju Guo ◽

Zhicheng Liu ◽

Minhua Sun ◽

...

Keyword(s):

Antiviral Activity ◽

Porcine Epidemic Diarrhea Virus ◽

Short Communication ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea

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Porcine Epidemic Diarrhea Virus (PEDV) ORF3 Enhances Viral Proliferation by Inhibiting Apoptosis of Infected Cells

Viruses ◽

10.3390/v12020214 ◽

2020 ◽

Vol 12 (2) ◽

pp. 214 ◽

Cited By ~ 3

Author(s):

Fusheng Si ◽

Xiaoxia Hu ◽

Chenyang Wang ◽

Bingqing Chen ◽

Ruiyang Wang ◽

...

Keyword(s):

Porcine Epidemic Diarrhea Virus ◽

Underlying Mechanism ◽

Vero Cells ◽

Orf3 Gene ◽

Accessory Gene ◽

Orf3 Protein ◽

Diarrhea Virus ◽

Viral Virulence ◽

Porcine Epidemic Diarrhea

The genomes of coronaviruses carry accessory genes known to be associated with viral virulence. The single accessory gene of porcine epidemic diarrhea virus (PEDV), ORF3, is dispensable for virus replication in vitro, while viral mutants carrying ORF3 truncations exhibit an attenuated phenotype of which the underlying mechanism is unknown. Here, we studied the effect of ORF3 deletion on the proliferation of PEDV in Vero cells. To this end, four recombinant porcine epidemic diarrhea viruses (PEDVs) were rescued using targeted RNA recombination, three carrying the full-length ORF3 gene from different PEDV strains, and one from which the ORF3 gene had been deleted entirely. Our results showed that PEDVs with intact or naturally truncated ORF3 replicated to significantly higher titers than PEDV without an ORF3. Further characterization revealed that the extent of apoptosis induced by PEDV infection was significantly lower with the viruses carrying an intact or C-terminally truncated ORF3 than with the virus lacking ORF3, indicating that the ORF3 protein as well as its truncated form interfered with the apoptosis process. Collectively, we conclude that PEDV ORF3 protein promotes virus proliferation by inhibiting cell apoptosis caused by virus infection. Our findings provide important insight into the role of ORF3 protein in the pathogenicity of PEDV.

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Investigation of the Role of the Spike Protein in Reversing the Virulence of the Highly Virulent Taiwan Porcine Epidemic Diarrhea Virus Pintung 52 Strains and Its Attenuated Counterpart

Viruses ◽

10.3390/v12010041 ◽

2019 ◽

Vol 12 (1) ◽

pp. 41 ◽

Cited By ~ 2

Author(s):

Chi-Fei Kao ◽

Hui-Wen Chang

Keyword(s):

Mortality Rate ◽

Porcine Epidemic Diarrhea Virus ◽

Rational Design ◽

Economic Losses ◽

Diarrhea Virus ◽

Viral Shedding ◽

S Gene ◽

Porcine Epidemic Diarrhea ◽

Highly Virulent

Porcine epidemic diarrhea virus (PEDV) has continuously caused severe economic losses to the global swine industries; however, no successful vaccine against PEDV has been developed. In this study, we generated four autologous recombinant viruses, including the highly virulent iPEDVPT-P5, attenuated iPEDVPT-P96, and two chimeric viruses (iPEDVPT-P5-96S and iPEDVPT-P96-5S) with the reciprocally exchanged spike (S) gene, to study the role of the S gene in PEDV pathogenesis. A deeper understanding of PEDV attenuation will aid in the rational design of a live attenuated vaccine (LAV) using reverse genetics system. Our results showed that replacing the S gene from the highly virulent iPEDVPT-P5 led to complete restoration of virulence of the attenuated iPEDVPT-P96, with nearly identical viral shedding, diarrhea pattern, and mortality rate as the parental iPEDVPT-P5. In contrast, substitution of the S gene with that from the attenuated iPEDVPT-P96 resulted in partial attenuation of iPEDVPT-P5, exhibiting similar viral shedding and diarrhea patterns as the parental iPEDVPT-P96 with slightly severe histological lesions and higher mortality rate. Collectively, our data confirmed that the attenuation of the PEDVPT-P96 virus is primarily attributed to mutations in the S gene. However, mutation in S gene alone could not fully attenuate the virulence of iPEDVPT-P5. Gene (s) other than S gene might also play a role in determining virulence.

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Effective inhibition of porcine epidemic diarrhea virus by RNA interference in vitro

Virus Genes ◽

10.1007/s11262-015-1242-5 ◽

2015 ◽

Vol 51 (2) ◽

pp. 252-259 ◽

Cited By ~ 6

Author(s):

Haiyan Shen ◽

Chunhong Zhang ◽

Pengju Guo ◽

Zhicheng Liu ◽

Jianfeng Zhang

Keyword(s):

Rna Interference ◽

Porcine Epidemic Diarrhea Virus ◽

Diarrhea Virus ◽

Effective Inhibition ◽

Porcine Epidemic Diarrhea

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Porcine Intestinal Enteroids: a New Model for Studying Enteric Coronavirus Porcine Epidemic Diarrhea Virus Infection and the Host Innate Response

Journal of Virology ◽

10.1128/jvi.01682-18 ◽

2018 ◽

Vol 93 (5) ◽

Cited By ~ 12

Author(s):

Liang Li ◽

Fang Fu ◽

Shanshan Guo ◽

Hongfeng Wang ◽

Xijun He ◽

...

Keyword(s):

Stem Cells ◽

Intestinal Epithelium ◽

Porcine Epidemic Diarrhea Virus ◽

Enteric Viruses ◽

Antiviral Response ◽

Diarrhea Virus ◽

Porcine Epidemic Diarrhea ◽

Ifn Lambda

ABSTRACTPorcine epidemic diarrhea virus (PEDV), a member of the group of alphacoronaviruses, is the pathogen of a highly contagious gastrointestinal swine disease. The elucidation of the events associated with the intestinal epithelial response to PEDV infection has been limited by the absence of goodin vitroporcine intestinal models that recapitulate the multicellular complexity of the gastrointestinal tract. Here, we generated swine enteroids from the intestinal crypt stem cells of the duodenum, jejunum, or ileum and found that the generated enteroids are able to satisfactorily recapitulate the complicated intestinal epitheliumin vivoand are susceptible to infection by PEDV. PEDV infected multiple types of cells, including enterocytes, stem cells, and goblet cells, and exhibited segmental infection discrepancies compared with ileal enteroids and colonoids, and this finding was verifiedin vivo. Moreover, the clinical isolate PEDV-JMS propagated better in ileal enteroids than the cell-adapted isolate PEDV-CV777, and PEDV infection suppressed interferon (IFN) production early during the infection course. IFN lambda elicited a potent antiviral response and inhibited PEDV in enteroids more efficiently than IFN alpha (IFN-α). Therefore, swine enteroids provide a novelin vitromodel for exploring the pathogenesis of PEDV and for thein vitrostudy of the interplay between a host and a variety of swine enteric viruses.IMPORTANCEPEDV is a highly contagious enteric coronavirus that causes significant economic losses, and the lack of a goodin vitromodel system is a major roadblock to an in-depth understanding of PEDV pathogenesis. Here, we generated a porcine intestinal enteroid model for PEDV infection. Utilizing porcine intestinal enteroids, we demonstrated that PEDV infects multiple lineages of the intestinal epithelium and preferably infects ileal enteroids over colonoids and that enteroids prefer to respond to IFN lambda 1 over IFN-α. These events recapitulate the events that occurin vivo. This study constitutes the first use of a primary intestinal enteroid model to investigate the susceptibility of porcine enteroids to PEDV and to determine the antiviral response following infection. Our study provides important insights into the events associated with PEDV infection of the porcine intestine and provides a valuablein vitromodel for studying not only PEDV but also other swine enteric viruses.

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