scholarly journals Identification and characterization of a novel linear epitope in the spike protein of the porcine epidemic diarrhea virus

2019 ◽  
Vol 63 (01) ◽  
pp. 88-95
Author(s):  
L. GONG ◽  
Y. GONG ◽  
Y. LIN ◽  
J. QIN ◽  
Y. LIU ◽  
...  
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Spike Protein ◽  
Linear Epitope ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea ◽  
Identification And Characterization

scholarly journals Characterization of influenza A virus pseudotyped with the spike protein of porcine epidemic diarrhea virus

2018 ◽  
Vol 163 (12) ◽  
pp. 3255-3264 ◽  
Author(s):  
Asawin Wanitchang ◽  
Janya Saenboonrueng ◽  
Kanjana Srisutthisamphan ◽  
Anan Jongkaewwattana
Keyword(s):  
Influenza A Virus ◽  
Porcine Epidemic Diarrhea Virus ◽  
Influenza A ◽  
Spike Protein ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

scholarly journals Cell Attachment Domains of the Porcine Epidemic Diarrhea Virus Spike Protein Are Key Targets of Neutralizing Antibodies

2017 ◽  
Vol 91 (12) ◽  
Author(s):  
Chunhua Li ◽  
Wentao Li ◽  
Eduardo Lucio de Esesarte ◽  
Hongbo Guo ◽  
Paul van den Elzen ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Receptor Binding ◽  
Porcine Epidemic Diarrhea Virus ◽  
Neutralizing Antibodies ◽  
Cell Attachment ◽  
Cross Reactivity ◽  
Spike Protein ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

ABSTRACT Porcine epidemic diarrhea virus (PEDV) causes enteric disease in pigs, resulting in significant economic losses to the swine industry worldwide. Current vaccination approaches against this emerging coronavirus are only partially effective, though natural infection protects pigs against reinfection and provides lactogenic immunity to suckling piglets. The viral spike (S) glycoprotein, responsible for receptor binding and cell entry, is the major target for neutralizing antibodies. However, knowledge of antibody epitopes, their nature and location in the spike structure, and the mechanisms by which the antibodies interfere with infection is scarce. Here we describe the generation and characterization of 10 neutralizing and nonneutralizing mouse monoclonal antibodies raised against the S1 receptor binding subunit of the S protein. By expression of different S1 protein fragments, six antibody epitope classes distributed over the five structural domains of the S1 subunit were identified. Characterization of antibodies for cross-reactivity and cross-neutralization revealed antigenic differences among PEDV strains. The epitopes of potent neutralizing antibodies segregated into two epitope classes and mapped within the N-terminal sialic acid binding domain and in the more C-terminal receptor binding domain. Antibody neutralization escape mutants displayed single amino acid substitutions that impaired antibody binding and neutralization and defined the locations of the epitopes. Our observations picture the antibody epitope landscape of the PEDV S1 subunit and reveal that its cell attachment domains are key targets of neutralizing antibodies. IMPORTANCE Porcine epidemic diarrhea virus (PEDV), an emerging porcine coronavirus, causes an economically important enteric disease in pigs. Effective PEDV vaccines for disease control are currently lacking. The spike (S) glycoprotein on the virion surface is the key player in virus cell entry and, therefore, the main target of neutralizing antibodies. To understand the antigenic landscape of the PEDV spike protein, we developed monoclonal antibodies against the spike protein's S1 receptor binding region and characterized their epitopes, neutralizing activity, and cross-reactivity toward multiple PEDV strains. Epitopes of antibodies segregated into six epitope classes dispersed over the multidomain S1 structure. Monoclonal antibodies revealed antigenic variability in B-cell epitopes between PEDV strains. The epitopes of neutralizing antibodies mapped to two distinct domains in S1 that are involved in binding to carbohydrate and proteinaceous cell surface molecules, respectively, indicating the importance of these cell attachment sites on the PEDV spike protein in eliciting a protective humoral immune response.

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

Virology ◽  
2017 ◽  
Vol 500 ◽  
pp. 50-61 ◽  
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

scholarly journals Engineering a Live Attenuated Porcine Epidemic Diarrhea Virus Vaccine Candidate via Inactivation of the Viral 2'-O-Methyltransferase and the Endocytosis Signal of the Spike Protein

2019 ◽  
Vol 93 (15) ◽  
Author(s):  
Yixuan Hou ◽  
Hanzhong Ke ◽  
Jineui Kim ◽  
Dongwan Yoo ◽  
Yunfang Su ◽  
...  
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Vaccine Development ◽  
Vaccine Candidate ◽  
Spike Protein ◽  
Economic Losses ◽  
High Dose ◽  
Type I ◽  
Viral Pathogen ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

ABSTRACT Porcine epidemic diarrhea virus (PEDV) causes high mortality in neonatal piglets; however, effective and safe vaccines are still not available. We hypothesized that inactivation of the 2′-O-methyltransferase (2′-O-MTase) activity of nsp16 and the endocytosis signal of the spike protein attenuates PEDV yet retains its immunogenicity in pigs. We generated a recombinant PEDV, KDKE4A, with quadruple alanine substitutions in the catalytic tetrad of the 2′-O-MTase using a virulent infectious cDNA clone, icPC22A, as the backbone. Next, we constructed another mutant, KDKE4A-SYA, by abolishing the endocytosis signal of the spike protein of KDKE4A. Compared with icPC22A, the KDKE4A and KDKE4A-SYA mutants replicated less efficiently in vitro but induced stronger type I and type III interferon responses. The pathogenesis and immunogenicities of the mutants were evaluated in gnotobiotic piglets. The virulence of KDKE4A-SYA and KDKE4A was significantly reduced compared with that of icPC22A. Mortality rates were 100%, 17%, and 0% in the icPC22A-, KDKE4A-, and KDKE4A-SYA-inoculated groups, respectively. At 21 days postinoculation (dpi), all surviving pigs were challenged orally with a high dose of icPC22A. The KDKE4A-SYA- and KDKE4A-inoculated pigs were protected from the challenge, because no KDKE4A-SYA- and one KDKE4A-inoculated pig developed diarrhea whereas all the pigs in the mock-inoculated group had severe diarrhea, and 33% of them died. Furthermore, we serially passaged the KDKE4A-SYA mutant in pigs three times and did not find any reversion of the introduced mutations. The data suggest that KDKE4A-SYA may be a PEDV vaccine candidate. IMPORTANCE PEDV is the most economically important porcine enteric viral pathogen and has caused immense economic losses in the pork industries in many countries. Effective and safe vaccines are desperately required but still not available. 2′-O-MTase (nsp16) is highly conserved among coronaviruses (CoVs), and the inactivation of nsp16 in live attenuated vaccines has been attempted for several betacoronaviruses. We show that inactivation of both 2′-O-MTase and the endocytosis signal of the spike protein is an approach to designing a promising live attenuated vaccine for PEDV. The in vivo passaging data also validated the stability of the KDKE4A-SYA mutant. KDKE4A-SYA warrants further evaluation in sows and their piglets and may be used as a platform for further optimization. Our findings further confirmed that nsp16 can be a universal target for CoV vaccine development and will aid in the development of vaccines against other emerging CoVs.

scholarly journals Molecular characterization of the spike gene of the porcine epidemic diarrhea virus in Mexico, 2013–2016

Virus Genes ◽  
2017 ◽  
Vol 54 (2) ◽  
pp. 215-224 ◽  
Author(s):  
Rocío Lara-Romero ◽  
Luis Gómez-Núñez ◽  
José Luis Cerriteño-Sánchez ◽  
Laura Márquez-Valdelamar ◽  
Susana Mendoza-Elvira ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Porcine Epidemic Diarrhea Virus ◽  
Spike Gene ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

scholarly journals Passive immunity to porcine epidemic diarrhea virus following immunization of pregnant gilts with a recombinant orf virus vector expressing the spike protein

2018 ◽  
Vol 163 (9) ◽  
pp. 2327-2335 ◽  
Author(s):  
Lok R. Joshi ◽  
Faten A. Okda ◽  
Aaron Singrey ◽  
Mayara F. Maggioli ◽  
Tatiane C. Faccin ◽  
...  
Keyword(s):  
Porcine Epidemic Diarrhea Virus ◽  
Virus Vector ◽  
Spike Protein ◽  
Orf Virus ◽  
Passive Immunity ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea
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