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

Abstract

Interferon-inducible transmembrane protein 3 (IFITM3) inhibits the replication of multiple pathogenic viruses by blocking their entry. In this study, we constructed a shuttle plasmid, harboring human IFITM3. Thereafter, recombinant adenovirus rAd5-IFITM3 was obtained by co-transfection of the linearized viral backbone vector pAd5 and the shuttle plasmid. The results showed that human IFITM3 did not affect the assembly and morphogenesis of progeny adenovirus. Human IFITM3 can be expressed in both A549 and MDCK cells in a time dependent manner. Furthermore, cells infected with rAd5-IFITM3 at a multiplicity of infection (MOI) of 100 for 24 h were challenged with avian influenza virus (AIV) H5N1 at an MOI of 1 for 6, 12 and 24 h. Rates of H5N1 infection in rAd5-IFITM3 cells were significantly decreased at 24 h post-infection (hpi), in a time dependent manner, compared with that of wild type wtAd5-infected cells. The expressions of viral genes were significantly inhibited at transcriptional and translational levels at 6 and 12 hpi. These results suggest that IFITM3 can suppress H5N1 replication in the early stage of the infection, which may be used as a promise agent against H5N1 infection in vivo.

Current Progress on Host Antiviral Factor IFITMs

“…Further research shows that silencing IFITM3 of U2OS cells can strongly enhance the replication of IAV H1N1 (A/PR/8/34), and siRNA targeting IFITM3 has a significant effect on eliminating IFN-g mediated virus inhibition (2). We previously found that IFITM3 can suppress H5N1 replication in the early stage of the infection (10). Moreover, overexpression of human IFITM1, 2, or 3 inhibits replication of an IAV H1N1 subtype (A/PR/8/34) and H3N2 subtype (A/Udorn/72) (2,24,25).…”

“…It is reported that IFITM proteins could significantly inhibit IAV, West Nile virus (WNV), Ebola virus (EBOV), SARS coronavirus (SARS-CoV), vesicular pharyngitis virus (VSV), Rift Valley fever virus (RVFV), dengue virus (DENV), Semliki forest virus (SFV), Zika virus, Respiratory syncytial virus (RSV), human immunodeficiency virus-1 (HIV-1), hepatitis C virus (HCV), Reovirus, and other capsular or noncapsular RNA viruses (2)(3)(4)(5)(6)(7)(8)(9)(10). Besides this, IFITM proteins also exhibit antiviral activity against individual DNA viruses (5,11,12).…”

Role of the Host Genetic Susceptibility to 2009 Pandemic Influenza A H1N1

“…TLR and RLH differ in their cellular localization, ligand specificity, and downstream signaling pathways, suggesting that host cells have multiple defense mechanisms against viral infection. Among ISGs, 2′–5′-oligoadenylate synthetase (OAS) plays a critical role in antiviral immunity by synthesizing 2–5As, which induces RNA degradation by activating a latent RNase (RNase L) pathway [ 34 ].…”

Antiviral activity of canine interferon lambda 3 expressed using a recombinant adenovirus against canine coronavirus, canine parvovirus, and canine distemper virus

“…For all viruses, the earlier the cells were transduced by Ad-caIFNλ3, the greater the suppression of both viral genome and protein levels. This could be the result of earlier expression of canine IFNλ3 by the recombinant adenovirus (Du et al 2019 ). In a similar context, inhibitory effects were better in a dose-dependent manner, showing better prevention in the cells treated with an MOI of 100 than those treated with an MOI of 10.…”