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Construction, expression and antiviral activity analysis of recombinant adenovirus expressing human IFITM3 in vitro
Shouwen Du1, Yinyue Jiang2, Wang Xu2
1Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Military Veterinary Institute, Academy of Military Medical Sciences, Changchun 130112, China; 2nd Clinical Medical College of Jinan University, Shenzhen People's Hospital, Shenzhen 518020, China.
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.
Insights
Interferon-inducible transmembrane protein 3 (IFITM3) effectively inhibits avian influenza virus (AIV) H5N1 replication by blocking viral entry. This study demonstrates IFITM3
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Interferon-inducible transmembrane protein 3 (IFITM3) is known to restrict the entry of various pathogenic viruses.
- Understanding IFITM3's role in antiviral defense is crucial for developing new therapeutic strategies.
Purpose of the Study:
- To investigate the antiviral activity of human IFITM3 against avian influenza virus (AIV) H5N1.
- To assess the efficacy of recombinant adenovirus expressing IFITM3 (rAd5-IFITM3) in inhibiting H5N1 replication.
Main Methods:
- Construction of a shuttle plasmid containing human IFITM3 and generation of recombinant adenovirus rAd5-IFITM3.
- Expression analysis of IFITM3 in A549 and MDCK cells.
- Challenging cells with AIV H5N1 after infection with rAd5-IFITM3 or wild-type adenovirus (wtAd5).
Main Results:
- Human IFITM3 expression did not impede adenovirus assembly or morphogenesis.
- rAd5-IFITM3-infected cells showed significantly reduced H5N1 infection rates in a time-dependent manner.
- IFITM3 suppressed viral gene expression at transcriptional and translational levels during early H5N1 infection.
Conclusions:
- IFITM3 effectively suppresses early-stage H5N1 replication.
- IFITM3 holds promise as a therapeutic agent against H5N1 infection in vivo.
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