A Recombinant Adenovirus Expressing Ovine Interferon Tau Prevents Influenza Virus-Induced Lethality in Mice

Abstract

bOvine interferon tau (IFN-) is a unique type I interferon with low toxicity and a broad host range in vivo. We report the generation of a nonreplicative recombinant adenovirus expressing biologically active IFN-. Using the B6.A2G-Mx1 mouse model, we showed that single-dose intranasal administration of recombinant Ad5-IFN-can effectively prevent lethality and disease induced by highly virulent hv-PR8 influenza virus by activating the interferon response and preventing viral replication.T ype I interferons (IFN) are crucial inducers of the antiviral response that act by binding to their universally expressed receptor (IFNAR) and regulating the expression of a large set of genes known as interferon-stimulated genes (ISGs) (1). Collectively, these ISGs mediate the establishment of an antiviral state directly precluding or diminishing viral replication within the cell (1, 2). Additionally, IFN modulates the deployment of a correct adaptive antiviral immune response by acting on cells of the immune system. For these reasons, IFN has long been thought of as an important antiviral treatment option (3, 4). However, its use in clinical settings as a broad-spectrum antiviral agent has been delayed due to its short half-life and the toxicity associated with its systemic or sustained delivery (5-8).Therefore, vectors for a localized delivery and tailoring of the signaling properties of IFN are important goals. Recently, adenoviral vectors expressing type I IFN have been generated and shown to be efficient in the prevention of a number of viral diseases in vertebrate hosts, including SARS coronavirus infections in ferrets and Ebola virus infection in macaques (9-12). IFN-is a type I interferon expressed in ruminants only which is known to induce low toxicity while maintaining its antiviral capacities (6,(13)(14)(15)(16)(17)(18). In contrast to other type I IFNs, IFN-can act on receptors from a broad range of species (17,19), and it has shown efficacy in reducing replication of different viruses, including human and feline immunodeficiency viruses (HIV and FIV), human papillomavirus (HPV) (7,20,21), and bovine leukemia virus (BKV) (8).Thus, we generated a recombinant nonreplicative second-generation human adenovirus (method previously described [22]), termed Ad5-IFN-, as a vector for the local delivery of IFN-in vivo (23). We first tested the ability of Ad5-IFN-to drive the expression and secretion of active IFN-. To do so, we assayed increasing amounts of medium from Vero cells, which lack endogenous type I IFN genes, infected with Ad5-IFN-or the parental control Ad5-DsRed virus and harvested at 48 h postinfection (hpi) in an antiviral activity assay. Vero cells were protected in a dose-dependent manner from vesicular stomatitis virus (VSV) infection when incubated with different amounts of the conditioned medium from Ad5-IFN--infected cells (Fig. 1A), indicating the presence of a secreted antiviral activity most likely corresponding to IFN-. As expected, control medium did not prevent VSV infection and showed no toxicity to...

Type I Interferons: Distinct Biological Activities and Current Applications for Viral Infection

“…Studies show that single-dose intranasal administration of recombinant Ad5-IFN-τ effectively protects mice from mortality and disease from the highly virulent hv-PR8 influenza virus. The mechanism of protection is inducing specific and time-dependent phosphorylation of STAT1, increasing mRNA for ISG15, MX1, and OAS1 and preventing viral replication [44]. …”

Construction, expression and antiviral activity analysis of recombinant adenovirus expressing human IFITM3 in vitro

“…Thus, a suitable vector or strategy is needed to transfer the protein into the target cell. Till now, adenovirus is one of the most effective means of delivering gene in vitro and in vivo, which have been widely used in therapies for cancer, highlevel, short-term expression of the target gene for gene therapy, vaccine for infection disease [1,2,5,[9][10][11][12][39][40][41][42]. In additionally, adenovirus vector is one of the most common vectors used in clinical trials, as human therapy studies based on adenovirus vector is up to 23.5% of all gene therapy trials worldwide [5].…”

“…The recombinant adenovirus rescued by transfecting adenovirus vectors into HEK293 cells are replication defective, with certain essential viral genes are replaced by a foreign gene [1,6], and the recombinant adenovirus can efficiently accommodate a larger foreign gene (up to 37 kb of foreign gene) and infect almost all types of cells and tissues [6][7][8]. Therefore, to date, replicationdefective recombinant Ad5 vectors have been considered as potential vaccine delivery vehicles for human immunodeficiency virus (HIV)-1, hepatitis C virus (HCV), influenza virus, and other viruses [9][10][11][12].…”

Virus‐induced autophagic degradation of STAT 2 as a mechanism for interferon signaling blockade

“…293T (ATCC CRL-1573), BHK-21 (ATCC CRL-6281C), SSC [65], and Vero cells (ATCC CCL-81) were grown in Dulbecco's modified Eagle medium (Invitrogen) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 1% 100× non-essential amino acids, 1 mM sodium pyruvate, and 100 U/ml penicillin/100 lg/ml streptomycin. All cells were kept at 37°C in the presence of 5% CO 2 .…”