Dataset Information

Species Specific Antiviral Activity of Porcine Interferon-?8 (IFN?8).

ABSTRACT: Interferons (IFNs) have been known as antiviral genes and they are classified by type 1, type 2, and type 3 IFN. The type 1 IFN consists of IFN?, IFN?, IFN?, and IFN? whereas the type 2 IFN consists of only IFN?, which is a key cytokine driving T helper cell type 1 immunity. IFN? belongs to the type 3 IFN, which is also known as IL-28 and IL-29 possessing antiviral activities. Type 1 IFN is produced by viral infection whereas type 2 IFN is induced by mitogenic or antigenic T-cell stimuli. The IFN? of bovine was first discovered in an ungulate ruminant recognition hormone. IFN? belongs to the type 1 IFN with the common feature of type 1 IFN such as antiviral activity. IFNs have been mostly studied for basic research and clinical usages therefore there was no effort to investigate IFNs in industrial animals. Here we cloned porcine IFN?8 from peripheral blood mononuclear cells of Korean domestic pig (Sus scrofa domestica). The newly cloned IFN?8 amino acid sequence from Korean domestic pig shares 98.4% identity with the known porcine IFN?8 in databank. The recombinant porcine IFN?8 showed potent antiviral activity and protected bovine Madin-Darby bovine kidney epithelial (MDBK) cells from the cytopathic effect of vesicular stomatitis virus, but it failed to protect human Wistar Institute Susan Hayflick (WISH) cells and canine Madin-Darby canine kidney epithelial-like (MDCK) cells. The present study demonstrates species specific antiviral activity of porcine IFN?8.

SUBMITTER: Kim E

PROVIDER: S-EPMC5746612 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

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Species Specific Antiviral Activity of Porcine Interferon-α8 (IFNα8).

Kim Eunhye E Jhun Hyunjhung H Kim Joohee J Park Unjoo U Jo Seunghyun S Kwak Areum A Kim Sinae S Nguyen Tam T TT Kang Yongsun Y Choi Insoo I Lee Joongbok J Kim Heijun H Kim Younghyun Y Lee Siyoung S Kim Soohyun S

Immune network 20171219 6

Interferons (IFNs) have been known as antiviral genes and they are classified by type 1, type 2, and type 3 IFN. The type 1 IFN consists of IFNα, IFNβ, IFNτ, and IFNω whereas the type 2 IFN consists of only IFNγ, which is a key cytokine driving T helper cell type 1 immunity. IFNλ belongs to the type 3 IFN, which is also known as IL-28 and IL-29 possessing antiviral activities. Type 1 IFN is produced by viral infection whereas type 2 IFN is induced by mitogenic or antigenic T-cell stimuli. The IF ...[more]

PMID: 29302255

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Project description:Type I interferons (IFNs), key antiviral cytokines, evolve to adapt with ever-changing viral threats during vertebrate speciation. Due to novel pathogenic pressure associated with Suidae speciation and domestication, porcine IFNs evolutionarily engender both molecular and functional diversification, which have not been well addressed in pigs, an important livestock species and animal model for biomedical sciences. Annotation of current swine genome assembly Sscrofa10.2 reveals 57 functional genes and 16 pseudogenes of type I IFNs. Subfamilies of multiple IFNA, IFNW and porcine-specific IFND genes are separated into four clusters with ∼ 60 kb intervals within the IFNB/IFNE bordered region in SSC1, and each cluster contains mingled subtypes of IFNA, IFNW and IFND. Further curation of the 57 functional IFN genes indicates that they include 18 potential artifactual duplicates. We performed phylogenetic construction as well as analyses of gene duplication/conversion and natural selection and showed that porcine type I IFN genes have been undergoing active diversification through both gene duplication and conversion. Extensive analyses of the non-coding sequences proximal to all IFN coding regions identified several genomic repetitive elements significantly associated with different IFN subtypes. Family-wide studies further revealed their molecular diversity with respect to differential expression and restrictive activity on the resurgence of a porcine endogenous retrovirus. Based on predicted 3-D structures of representative animal IFNs and inferred activity, we categorized the general functional propensity underlying the structure-activity relationship. Evidence indicates gene expansion of porcine type I IFNs. Genomic repetitive elements that associated with IFN subtypes may serve as molecular signatures of respective IFN subtypes and genomic mechanisms to mediate IFN gene evolution and expression. In summary, the porcine type I IFN profile has been phylogenetically defined family-wide and linked to diverse expression and antiviral activity, which is important information for further biological studies across the porcine type I IFN family.

Project description:We determined the interferon-gamma (IFN-gamma) cDNA sequence from three porcine breeds, Duroc, Landrance/Duroc hybrid, and Landrance breeds. Five single nucleotide polymorphisms (SNPs) of porcine IFN-gamma (PoIFN-gamma) were identified, respectively, at positions 269 (A/G), 376 (C/T), 426 (T/C), and 465 (T/C) of the coding sequence in Landrance/Duroc hybrid, and at position 251 (A/G) in Landrance breed. Among them, A269G and A251G polymorphisms resulted in Q67R and K61R replacements in the mature protein. PoIFN-gamma cDNAs of Duroc breed (PoIFN-gamma-W) and Landrance/Duroc hybrid (PoIFN-gamma-M), which, respectively, encoded Q67 and R67, were introduced into a prokaryotic expression vector pET32 to express recombinant PoIFN-gamma-W (rPoIFN-gamma-W) and rPoIFN-gamma-M protein variants in Escherichia coli. The identity of both protein variants was further confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). We then compared bioactivities of these two recombinant proteins. Although both recombinant protein variants exhibited comparable activities in antiproliferation of PK-15 cells and in nitric oxide (NO) induction of porcine peripheral monocytes, antiviral activity of rPoIFN-gamma-W protein was significantly higher (P<0.001) than that of rPoIFN-gamma-M protein in a plaque inhibition assay using pseudorabies virus (PRV). IC50 values of rPoIFN-gamma-W and rPoIFN-gamma-M protein in anti-PRV assay were determined as 5.3+/-1.3 and 9.3+/-4.3nM, respectively. In conclusion, we have identified five novel SNPs in PoIFN-gamma cDNA, including two missense polymorphisms that result in Q67R and K61R replacements. Our results further demonstrate that Q67R can markedly reduce antiviral activity of the PoIFN-gamma protein. This is the first report that shows the functional SNP in the coding region of IFN-gamma. In the future, it is imperative to determine whether Q67R replacement in IFN-gamma may have disease association.

Dataset Information

Species Specific Antiviral Activity of Porcine Interferon-?8 (IFN?8).

Publications

Species Specific Antiviral Activity of Porcine Interferon-α8 (IFNα8).

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