Molecular Basis for Broad Neuraminidase Immunity: Conserved Epitopes in Seasonal and Pandemic H1N1 as Well as H5N1 Influenza Viruses

Wan, Hongquan; Gao, Jin; Xu, Kemin; Chen, Hongjun; Couzens, Laura; Rivers, Katie; Easterbrook, Judy; Yang, Kevin Yi; Zhong, Lei; Rajabi, Mohsen; Ye, Jianqiang; Sultana, Ishrat; Wan, Xiu‐Feng; Liu, Xiufan; Pérez, Daniel R.; Taubenberger, Jeffery K.; Eichelberger, Maryna C.

doi:10.1128/jvi.01203-13

Cited by 136 publications

(173 citation statements)

References 46 publications

(61 reference statements)

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“…A cell-based ELISA was performed as previously described (27). NA was expressed on 293T cells by transfection with wt or mutant plasmids using the Lipofectamine 2000 reagent (Invitrogen Inc., Grand Island, NY).…”

Section: Methodsmentioning

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%

“…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%

“…A universal epitope in NA has been reported, and a rabbit MAb against this epitope was shown to inhibit multiple subtypes of NA (25,26), although it remains to be investigated whether this epitope elicits an immune response in vivo. We recently defined the NA antigenic domains of a seasonal H1N1 virus, A/Brisbane/59/2007 (BR/07), by identifying amino acids in NA that are critical for binding of a panel of MAbs (27). These included a group of MAbs that are highly specific for the NA of BR/07-like viruses (referred to as group A) and a group of MAbs that are broadly reactive with N1 subtypes (referred to as group B), including the NA of a 2009 pH1N1 virus, A/California/7/2009 (CA/09).…”

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: Methodsmentioning

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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show abstract

“…Antibodies against the other major viral surface glycoprotein, the NA, have long been ignored, but recent reports show that NA-based vaccines are able to protect from disease as well -mainly by inhibiting the enzymatic activity of the NA [18][19][20]. This neuraminidase inhibiting (NI) activity might be a critical factor in the absence of neutralizing HI active antibodies (e.g., in the case of H7 vaccines) but might also contribute to protection induced by seasonal influenza virus vaccines.…”

mentioning

confidence: 99%

The emergence of H7N9 viruses: a chance to redefine correlates of protection for influenza virus vaccines

Krammer¹

2013

Expert Review of Vaccines

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Diagnostics and surveillance methods

2016

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Molecular Basis for Broad Neuraminidase Immunity: Conserved Epitopes in Seasonal and Pandemic H1N1 as Well as H5N1 Influenza Viruses

Cited by 136 publications

References 46 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

The emergence of H7N9 viruses: a chance to redefine correlates of protection for influenza virus vaccines

Diagnostics and surveillance methods

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