Cloning, expression and antiviral activity of mink alpha-interferons

Abstract

BackgroundAs a key link between innate and adaptive immune responses, the interferon (IFN) system is the first line of defense against viral infection. IFN, and in particular, IFN-α, has been used clinically as an effective therapeutic agent for viral infections. However, different subtypes of IFN-α demonstrate distinct antiviral activity. Therefore, it is important to identify IFN-α subtypes with high antiviral activity for the development of genetically engineered antiviral drugs.ResultsIn this study, we cloned the genes for 13 IFN-α subtypes from peripheral blood lymphocytes of the mink. The homologies of the 13 mink IFN-α genes were 93.6–99.3% and 88.8–98.4% at the nucleotide and amino acid sequence levels, respectively. In contrast to human and canine IFN-α subtypes, most mink IFN-α subtypes contained two N-glycosylation sites. We expressed and purified 13 mink IFN-α subtypes in Escherichia coli. The cytopathic effect inhibition assay showed that all the 13 recombinant mink IFN-α subtypes inhibited the propagation of vesicular stomatitis virus in WISH cells, with IFN-α2 and IFN-α12 demonstrating the highest activities. Furthermore, recombinant mink IFN-α2 and IFN-α12 significantly suppressed the propagation of canine distemper virus in Vero cells, with IFN-α2 demonstrating the highest activity.ConclusionsWe identified the mink IFN-α2 subtype as a promising candidate for the development of effective antiviral drugs.

Cloning and expression of mink (Neovison vison) interferon-γ gene and development of an antiviral assay

“…In recent years, mink distemper, mink parvovirus enteritis, aeutian disease and other infectious diseases have been endangering the survival of mink Zhang et al, 2015). Thus, the prevention and treatment of these diseases cannot be ignored.…”

Antiviral activity of mink interferon alpha expressed in the yeast Pichia pastoris

“…Imidazole was removed by dialysis against PBS. Western blotting of the partially purified 6 × His-tagged IFN-α was performed as described previously ( 10 ). The purified protein was quantified with the bicinchoninic acid (BCA) protein assay kit using bovine serum albumin as a standard.…”

“…Blood was collected from the tail vein at 0 h, 15 min, 30 min, and 1, 2, 4, 6, 8, 12, 24, 36, 48, 72, 96, 108, 132, and 156 h after injection, and serum was obtained by centrifugation at 4,000 rpm for 20 min at 4°C. As no commercial mink IFN-α detection kit was available, and mink and canine IFN-α share 80.9% homology at the nucleotide sequence ( 10 ), serum MiIFN-α was quantified using canine IFN alpha-1/2 (IFN-α1/2) ELISA (Abebio, China). Western blot experiments confirmed that canine IFN-α monoclonal antibody can cross-react with mink IFN-α, and thus, we used the commercial canine IFN-α ELISA kit to detect mink IFN-α in rat serum.…”

“…Mink IFN-α is a protein family encoded by 13 functional genes, which share similar lengths and amino acid sequence homology of 88.8–98.4%. The mature MiIFN-α contains 167 amino acids, and the signal peptide comprises 23 amino acids ( 10 ). Nonetheless, as yet, there has been no domestic production of genetically engineered mink IFN antiviral agents or the joint use of related products.…”

Temporal Dynamics of the Ruminant Type I IFN-Induced Antiviral State against Homologous Parainfluenza Virus 3 Challenge In Vitro

“…Currently, the IFN-α proteins encoded by humans and other animal species are all highly effective in limiting virus replication and spread [ 4 , 5 , 6 , 7 ]. Although IFNs are broad-spectrum antiviral agents, the sequence conservation of the IFN-α proteins from different species can differ considerably [ 8 , 9 ], and there are also differences in the potency of different IFN-a homologues [ 10 , 11 ]. The antiviral property of goat IFN-α has not been evaluated for prophylaxis and/or therapeutic treatment of viral infections to date.…”