mRNA Encoding a Bispecific Single Domain Antibody Construct Protects against Influenza A Virus Infection in Mice

ABSTRACTAlpaca-derived single-domain antibody fragments (VHHs) that target the influenza A virus nucleoprotein (NP) can protect cells from infection when expressed in the cytosol. We found that one such VHH, αNP-VHH1, exhibits antiviral activity similar to that of Mx proteins by blocking nuclear import of incoming viral ribonucleoproteins (vRNPs) and viral transcription and replication in the nucleus. We determined a 3.2-Å crystal structure of αNP-VHH1 in complex with influenza A virus NP. The VHH binds to a nonconserved region on the body domain of NP, which has been associated with binding to host factors and serves as a determinant of host range. Several of the NP/VHH interface residues determine sensitivity of NP to antiviral Mx GTPases. The structure of the NP/αNP-VHH1 complex affords a plausible explanation for the inhibitory properties of the VHH and suggests a rationale for the antiviral properties of Mx proteins. Such knowledge can be leveraged for much-needed novel antiviral strategies.IMPORTANCEInfluenza virus strains can rapidly escape from protection afforded by seasonal vaccines or acquire resistance to available drugs. Additional ways to interfere with the virus life cycle are therefore urgently needed. The influenza virus nucleoprotein is one promising target for antiviral interventions. We have previously isolated alpaca-derived single-domain antibody fragments (VHHs) that protect cells from influenza virus infection if expressed intracellularly. We show here that one such VHH exhibits antiviral activities similar to those of proteins of the cellular antiviral defense (Mx proteins). We determined the three-dimensional structure of this VHH in complex with the influenza virus nucleoprotein and identified the interaction site, which overlaps regions that determine sensitivity of the virus to Mx proteins. Our data define a new vulnerability of influenza virus, help us to better understand the cellular antiviral mechanisms, and provide a well-characterized tool to further study them.

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Potent Neutralization of Influenza A Virus by a Single-Domain Antibody Blocking M2 Ion Channel Protein

PLoS ONE ◽

10.1371/journal.pone.0028309 ◽

2011 ◽

Vol 6 (12) ◽

pp. e28309 ◽

Cited By ~ 26

Author(s):

Guowei Wei ◽

Weixu Meng ◽

Haijiang Guo ◽

Weiqi Pan ◽

Jinsong Liu ◽

...

Keyword(s):

Ion Channel ◽

Influenza A Virus ◽

Influenza A ◽

Single Domain ◽

Single Domain Antibody ◽

Channel Protein ◽

Domain Antibody ◽

Potent Neutralization

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Faculty Opinions recommendation of Enhanced susceptibility of Ago1/3 double-null mice to influenza A virus infection.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.717948803.793453903 ◽

2012 ◽

Author(s):

Kuan-Teh Jeang

Keyword(s):

Virus Infection ◽

Influenza A Virus ◽

Influenza A ◽

Influenza A Virus Infection

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Faculty Opinions recommendation of Effect of 1918 PB1-F2 expression on influenza A virus infection kinetics.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.8733956.9247056 ◽

2011 ◽

Author(s):

Bryan Grenfell ◽

Nimalan Arinaminpathy

Keyword(s):

Virus Infection ◽

Influenza A Virus ◽

Influenza A ◽

Influenza A Virus Infection

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Intractable Ventricular Fibrillation Following Influenza A Virus Infection

10.1164/ajrccm-conference.2019.199.1_meetingabstracts.a6583 ◽

2019 ◽

Author(s):

S. Saleem ◽

E. Shah ◽

M. Corpuz ◽

M. Karwa

Keyword(s):

Ventricular Fibrillation ◽

Virus Infection ◽

Influenza A Virus ◽

Influenza A ◽

Influenza A Virus Infection

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Induction of cyclophilin A by influenza A virus infection facilitates group A Streptococcus coinfection

Cell Reports ◽

10.1016/j.celrep.2021.109159 ◽

2021 ◽

Vol 35 (7) ◽

pp. 109159

Author(s):

Xiaoyuan Bai ◽

Wenxian Yang ◽

Xiaohan Luan ◽

Huizi Li ◽

Heqiao Li ◽

...

Keyword(s):

Virus Infection ◽

Influenza A Virus ◽

Influenza A ◽

Group A Streptococcus ◽

Cyclophilin A ◽

Group A ◽

Influenza A Virus Infection

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Dicer is involved in protection against influenza A virus infection

Journal of General Virology ◽

10.1099/vir.0.83103-0 ◽

2007 ◽

Vol 88 (10) ◽

pp. 2627-2635 ◽

Cited By ~ 80

Author(s):

Alexey A. Matskevich ◽

Karin Moelling

Keyword(s):

Virus Infection ◽

Influenza A Virus ◽

Mammalian Cells ◽

Influenza A ◽

Virus Production ◽

Vero Cells ◽

Antiviral Response ◽

Protein Levels ◽

Infected Cells ◽

Influenza A Virus Infection

In mammals the interferon (IFN) system is a central innate antiviral defence mechanism, while the involvement of RNA interference (RNAi) in antiviral response against RNA viruses is uncertain. Here, we tested whether RNAi is involved in the antiviral response in mammalian cells. To investigate the role of RNAi in influenza A virus-infected cells in the absence of IFN, we used Vero cells that lack IFN-α and IFN-β genes. Our results demonstrate that knockdown of a key RNAi component, Dicer, led to a modest increase of virus production and accelerated apoptosis of influenza A virus-infected cells. These effects were much weaker in the presence of IFN. The results also show that in both Vero cells and the IFN-producing alveolar epithelial A549 cell line influenza A virus targets Dicer at mRNA and protein levels. Thus, RNAi is involved in antiviral response, and Dicer is important for protection against influenza A virus infection.

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