Structural characterization of a protective epitope spanning A(H1N1)pdm09 influenza virus neuraminidase monomers

Wan, Hongquan; Yang, Hua; Shore, D.A.; Garten, Rebecca; Couzens, Laura; Gao, Jin; Jiang, Lianlian; Carney, P.J.; Villanueva, Julie; Stevens, James; Eichelberger, Maryna C.

doi:10.1038/ncomms7114

Cited by 92 publications

(109 citation statements)

References 51 publications

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“…1A). The results of our previous NI assays suggest that HF5 may bind to an epitope involving residue 369 (28). In the present study, we examined the binding of HF5 to various mutant CA/09 NAs transiently expressed on 293T cells.…”

Section: Resultsmentioning

confidence: 99%

“…NA residues 95, 449, and 451 are key for the binding of CD6. Antibodies 4E9 and 1H5 are broadly reactive with the NA of H1N1 (including seasonal and pandemic H1N1 viruses) and H5N1 viruses, with NA residues 273, 338, and 339 being the critical contacts (27,28).…”

Section: Resultsmentioning

confidence: 99%

“…Virus counting was performed as reported previously (32) using a Virus Counter 2100 system (ViroCyt, Boulder, CO). Hybridomas that secrete MAbs were generated as previously described (27,28) and cultured in a CELLine device (BD Biosciences, San Jose, CA). MAbs were purified using protein G columns (GE Healthcare, Uppsala, Sweden) according to the instructions provided by the manufacturer.…”

Section: Methodsmentioning

confidence: 99%

“…The amino acids essential for binding group B antibodies are highly conserved in the N1 subtype of viruses, including H5N1. In a separate study, we characterized two MAbs, HF5 and CD6, that are specific to the NA of pH1N1 virus (28). When tested in vivo, CD6 and MAbs from group B protected DBA/2 mice against challenge with a lethal dose of reassortant vaccine candidate CA/09-X179A (27,28).…”

mentioning

confidence: 99%

“…In a separate study, we characterized two MAbs, HF5 and CD6, that are specific to the NA of pH1N1 virus (28). When tested in vivo, CD6 and MAbs from group B protected DBA/2 mice against challenge with a lethal dose of reassortant vaccine candidate CA/09-X179A (27,28). DBA/2 mice are sensitive to many subtypes of influenza viruses (29), permitting challenge experiments to be conducted with viruses that have a range of virulences.…”

mentioning

confidence: 99%

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Comparative Efficacy of Monoclonal Antibodies That Bind to Different Epitopes of the 2009 Pandemic H1N1 Influenza Virus Neuraminidase

Jiang

Fantoni

Couzens

et al. 2016

J Virol

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Antibodies against the neuraminidase (NA) of influenza virus correlate with resistance against disease, but the effectiveness of antibodies against different NA epitopes has not been compared. In the present study, we evaluated the in vitro and in vivo efficacies of four monoclonal antibodies (MAbs): HF5 and CD6, which are specific to two different epitopes in the NA of 2009 pandemic H1N1 (pH1N1) virus, and 4E9 and 1H5, which are specific to a conserved epitope in the NA of both H1N1 and H5N1 viruses. In the in vitro assays, HF5 and CD6 inhibited virus spread and growth more effectively than 4E9 and 1H5, with HF5 being the most effective inhibitor. When administered prophylactically at 5 mg/kg of body weight, HF5 and CD6 protected ϳ90 to 100% of DBA/2 mice against lethal wild-type pH1N1 virus challenge; however, at a lower dose (1 mg/kg), HF5 protected ϳ90% of mice, whereas CD6 protected only 25% of mice. 4E9 and 1H5 were less effective than HF5 and CD6, as indicated by the partial protection achieved even at doses as high as 15 mg/kg. When administered therapeutically, HF5 protected a greater proportion of mice against lethal pH1N1 challenge than CD6. However, HF5 quickly selected pH1N1 virus escape mutants in both prophylactic and therapeutic treatments, while CD6 did not. Our findings confirm the important role of NA-specific antibodies in immunity to influenza virus and provide insight into the properties of NA antibodies that may serve as good candidates for therapeutics against influenza. IMPORTANCENeuraminidase (NA) is one of the major surface proteins of influenza virus, serving as an important target for antivirals and therapeutic antibodies. The impact of NA-specific antibodies on NA activity and virus replication is likely to depend on where the antibody binds. Using in vitro assays and the mouse model, we compared the inhibitory/protective efficacy of four mouse monoclonal antibodies (MAbs) that bind to different sites within the 2009 pandemic H1N1 (pH1N1) virus NA. The ability of each MAb to protect mice against lethal pH1N1 infection corresponded to its ability to inhibit NA activity in vitro; however, the MAb that was the most effective inhibitor of NA activity selected pH1N1 escape variants in vivo. One of the tested MAbs, which binds to a conserved region in the NA of pH1N1 virus, inhibited NA activity but did not result in escape variants, highlighting its suitability for development as a therapeutic agent.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Comparative Efficacy of Monoclonal Antibodies That Bind to Different Epitopes of the 2009 Pandemic H1N1 Influenza Virus Neuraminidase

Jiang

Fantoni

Couzens

et al. 2016

J Virol

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Carbohydrate‐Based Antiviral Vaccines

Plata,

Fernández‐Tejada

2023

Carbohydrate‐Based Therapeutics

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Double‐Layered M2e‐NA Protein Nanoparticle Immunization Induces Broad Cross‐Protection against Different Influenza Viruses in Mice

Wang

Deng

Gonzalez

et al. 2019

Adv Healthcare Materials

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The development of a universal influenza vaccine is an ideal strategy to eliminate public health threats from influenza epidemics and pandemics. This ultimate goal is restricted by the low immunogenicity of conserved influenza epitopes. Layered protein nanoparticles composed of well‐designed conserved influenza structures have shown improved immunogenicity with new physical and biochemical features. Herein, structure‐stabilized influenza matrix protein 2 ectodomain (M2e) and M2e‐neuraminidase fusion (M2e‐NA) recombinant proteins are generated and M2e protein nanoparticles and double‐layered M2e‐NA protein nanoparticles are produced by ethanol desolvation and chemical crosslinking. Immunizations with these protein nanoparticles induce immune protection against different viruses of homologous and heterosubtypic NA in mice. Double‐layered M2e‐NA protein nanoparticles induce higher levels of humoral and cellular responses compared with their comprising protein mixture or M2e nanoparticles. Strong cytotoxic T cell responses are induced in the layered M2e‐NA protein nanoparticle groups. Antibody responses contribute to the heterosubtypic NA immune protection. The protective immunity is long lasting. These results demonstrate that double‐layered protein nanoparticles containing structure‐stabilized M2e and NA can be developed into a universal influenza vaccine or a synergistic component of such vaccines. Layered protein nanoparticles can be a general vaccine platform for different pathogens.

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Structural characterization of a protective epitope spanning A(H1N1)pdm09 influenza virus neuraminidase monomers

Cited by 92 publications

References 51 publications

Comparative Efficacy of Monoclonal Antibodies That Bind to Different Epitopes of the 2009 Pandemic H1N1 Influenza Virus Neuraminidase

Comparative Efficacy of Monoclonal Antibodies That Bind to Different Epitopes of the 2009 Pandemic H1N1 Influenza Virus Neuraminidase

Carbohydrate‐Based Antiviral Vaccines

Double‐Layered M2e‐NA Protein Nanoparticle Immunization Induces Broad Cross‐Protection against Different Influenza Viruses in Mice

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