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PMID: 17325366 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Recombinant swine beta interferon protects swine alveolar macrophages and MARC-145 cells from infection with Porcine reproductive and respiratory syndrome virus.

The Journal of general virology ·Vol. 88 ·No. Pt 3 ·2007-03-00 ·Pages 925-931

Overend C, Mitchell R, He D, Rompato G, Grubman MJ, Garmendia AE

Abstract

Swine beta interferon (swIFN-beta) produced in HEK 293 cells infected with a recombinant, replication-defective human adenovirus 5 (Ad5) encoding the swIFN-beta gene was tested for antiviral activity against Porcine reproductive and respiratory syndrome virus (PRRSV). MARC-145 cells were incubated overnight with dilutions of supernatant fluids from HEK 293 cells infected with Ad5-swIFN-beta or with an Ad5 control virus (Ad5-Blue). Treated cells were infected with PRRSV; MARC-145 cells incubated with Ad5-Blue supernatants developed cytopathic effects (CPE), whereas those incubated with swIFN-beta showed no CPE. To confirm the antiviral activity of swIFN-beta, culture fluids from Ad5-swIFN-beta-infected cells were affinity-purified on a Sepharose-anti-swIFN-beta matrix, and the resulting fractions exhibited antiviral activity upon infection with PRRSV. The antiviral effects were specific, as they were blocked by mAbs against swIFN-beta. Additional cultures of MARC-145 cells treated with swIFN-beta-containing supernatants or affinity-purified swIFN-beta were infected with PRRSV and tested by real-time RT-PCR for viral RNA in culture supernatants at various times post-inoculation. These experiments confirmed the protective effects of swIFN-beta. swIFN-beta was also tested for antiviral activity on porcine alveolar macrophages (PAMs) obtained by bronchoalveolar lavage from PRRSV-negative swine. PAMs were treated with dilutions of swIFN-beta or Ad5-Blue culture fluids, infected with PRRSV and tested for viral RNA by real-time RT-PCR. The viral load data showed a dose-dependent protection in swIFN-beta-treated PAMs, whereas no protection was evident from Ad5-Blue culture fluids. The data demonstrate that swIFN-beta protects both MARC-145 cells and PAMs from PRRSV infection.

MeSH Terms
Adenoviridae/genetics Animals Antibodies, Monoclonal/immunology Antiviral Agents/isolation & purification,pharmacology Cell Line Chromatography, Affinity Cytopathogenic Effect, Viral/drug effects Dose-Response Relationship, Drug Genetic Vectors Humans Interferon Type I/immunology,pharmacology Macrophages, Alveolar/physiology,virology Porcine respiratory and reproductive syndrome virus/drug effects,physiology RNA, Viral/biosynthesis Recombinant Proteins Reverse Transcriptase Polymerase Chain Reaction Swine Virus Replication/drug effects
Chemicals
Antibodies, Monoclonal Antiviral Agents Interferon Type I RNA, Viral Recombinant Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Overend C
Department of Pathobiology and Veterinary Science, University of Connecticut, 61 North Eagleville Road, Storrs, CT, USA.
Mitchell R
Department of Pathobiology and Veterinary Science, University of Connecticut, 61 North Eagleville Road, Storrs, CT, USA.
He D
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
Rompato G
Department of Pathobiology and Veterinary Science, University of Connecticut, 61 North Eagleville Road, Storrs, CT, USA.
Grubman M J
Plum Island Animal Disease Center, ARS, USDA, Greenport, NY, USA.
Garmendia A E
Department of Pathobiology and Veterinary Science, University of Connecticut, 61 North Eagleville Road, Storrs, CT, USA.
Article Info
Journal
The Journal of general virology
Abbr.
J Gen Virol
ISSN
0022-1317
Published
2007-03-00
Pages
925-931
Language
English
Region
England
NLM ID
0077340
Subset
IM
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