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. 2020 May 30;12(6):597.
doi: 10.3390/v12060597.

Antibody Neutralization of an Influenza Virus that Uses Neuraminidase for Receptor Binding

Lauren E Gentles  1   2 , Hongquan Wan  3 , Maryna C Eichelberger  4 , Jesse D Bloom  1   2   5
Affiliations

Antibody Neutralization of an Influenza Virus that Uses Neuraminidase for Receptor Binding

Lauren E Gentles et al. Viruses. .

Abstract

Influenza virus infection elicits antibodies against the receptor-binding protein hemagglutinin (HA) and the receptor-cleaving protein neuraminidase (NA). Because HA is essential for viral entry, antibodies targeting HA often potently neutralize the virus in single-cycle infection assays. However, antibodies against NA are not potently neutralizing in such assays, since NA is dispensable for single-cycle infection. Here we show that a modified influenza virus that depends on NA for receptor binding is much more sensitive than a virus with receptor-binding HA to neutralization by some anti-NA antibodies. Specifically, a virus with a receptor-binding G147R N1 NA and a binding-deficient HA is completely neutralized in single-cycle infections by an antibody that binds near the NA active site. Infection is also substantially inhibited by antibodies that bind NA epitopes distant from the active site. Finally, we demonstrate that this modified virus can be used to efficiently select mutations in NA that escape antibody binding, a task that can be laborious with typical influenza viruses that are not well neutralized by anti-NA antibodies. Thus, viruses dependent on NA for receptor binding allow for sensitive in vitro detection of antibodies binding near the catalytic site of NA and enable the selection of viral escape mutants.

Keywords: G147R; antibody escape; influenza virus; neuraminidase; neutralization; receptor-binding.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Proposed mechanism of neutralization of neuraminidase (NA)-binding-dependent viruses by anti-NA antibodies. Most influenza viruses use hemagglutinin (HA) to bind and enter cells (left panel), so antibodies targeting HA can prevent infection while NA antibodies only inhibit the release of new virions. But in our engineered NA-binding-dependent virus (right panel), antibodies against NA can inhibit infection.
Figure 2
Figure 2
Neutralization of viruses with different dependencies on HA and NA for receptor binding by anti-NA antibodies in single-cycle infection assays. Neutralization curves show the percentage of the NAwt/HAwt, NAbind/HAwt, or NAbind/HA∆bind virus infection that is inhibited by each antibody over a range of concentrations compared to the no-antibody control infection. Points represent the mean of three technical replicates with error bars showing the standard error of the mean.
Figure 3
Figure 3
S364N increases resistance to neutralization by antibody HF5 in single-cycle infections for viruses that are dependent on NA for receptor-binding. Results are shown as the percent infection at each concentration compared to the no-antibody infection control. Points represent the mean of three technical replicates with error bars showing the standard error of the mean.
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