Reverse Genetic Assessment of the Roles Played by the Spike Protein and ORF3 in Porcine Epidemic Diarrhea Virus Pathogenicity.

Kristen-Burmann, Claudia; Rogger, Peter; Berenguer Veiga, Inês; Riebesehl, Stefanie; Rappe, Julie; Ebert, Nadine; Sautter, Carmen A; Kelly, Jenna N; Stalder, Hanspeter; Ehmann, Rosina; Huber, Michael; Posthaus, Horst; Ruggli, Nicolas; Thiel, Volker; Tekes, Gergely (2023). Reverse Genetic Assessment of the Roles Played by the Spike Protein and ORF3 in Porcine Epidemic Diarrhea Virus Pathogenicity. Journal of virology, 97(7), e0196422. American Society for Microbiology 10.1128/jvi.01964-22

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Porcine epidemic diarrhea virus is a swine pathogen that has been responsible for significant animal and economic losses worldwide in recent years. In this manuscript, we report the generation of a reverse genetics system C(RGS) for the highly virulent US PEDV strain Minnesota (PEDV-MN; GenBank accession number KF468752), which was based on the assembly and cloning of synthetic DNA, using vaccinia virus as a cloning vector. Viral rescue was only possible following the substitution of 2 nucleotides within the 5'UTR and 2 additional nucleotides within the spike gene, based on the sequence of the cell culture-adapted strains. Besides displaying a highly pathogenic phenotype in newborn piglets, in comparison with the parental virus, the rescued recombinant PEDV-MN was used to confirm that the PEDV spike gene has an important role in PEDV virulence and that the impact of an intact PEDV ORF3 on viral pathogenicity is modest. Moreover, a chimeric virus with a TGEV spike gene in the PEDV backbone generated with RGS was able to replicate efficiently in vivo and could be readily transmitted between piglets. Although this chimeric virus did not cause severe disease upon the initial infection of piglets, there was evidence of increasing pathogenicity upon transmission to contact piglets. The RGS described in this study constitutes a powerful tool with which to study PEDV pathogenesis and can be used to generate vaccines against porcine enteric coronaviruses. IMPORTANCE PEDV is a swine pathogen that is responsible for significant animal and economic losses worldwide. Highly pathogenic variants can lead to a mortality rate of up to 100% in newborn piglets. The generation of a reverse genetics system for a highly virulent PEDV strain originating from the United States is an important step in phenotypically characterizing PEDV. The synthetic PEDV mirrored the authentic isolate and displayed a highly pathogenic phenotype in newborn piglets. With this system, it was possible to characterize potential viral virulence factors. Our data revealed that an accessory gene (ORF3) has a limited impact on pathogenicity. However, as it is also now known for many coronaviruses, the PEDV spike gene is one of the main determinants of pathogenicity. Finally, we show that the spike gene of another porcine coronavirus, namely, TGEV, can be accommodated in the PEDV genome background, suggesting that similar viruses can emerge in the field via recombination.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Animal Pathology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Virology and Immunology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)

UniBE Contributor:

Rogger, Peter, Berenguer Veiga, Inês Margarida, Rappe, Julie Christiane Françoise, Ebert, Nadine, Sautter, Carmen Alexandra Nadine, Kelly, Jenna Nicole, Stalder, Hanspeter, Posthaus, Horst, Ruggli, Nicolas, Thiel, Volker Earl

Subjects:

600 Technology > 630 Agriculture

ISSN:

0022-538X

Publisher:

American Society for Microbiology

Language:

English

Submitter:

Pubmed Import

Date Deposited:

27 Jun 2023 09:13

Last Modified:

28 Jul 2023 00:15

Publisher DOI:

10.1128/jvi.01964-22

PubMed ID:

37358450

Uncontrolled Keywords:

ORF3 porcine epidemic diarrhea virus spike gene synthesized DNA vaccinia virus reverse genetics

BORIS DOI:

10.48350/184161

URI:

https://boris.unibe.ch/id/eprint/184161

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