Safe Pseudovirus-based Assay for Neutralization Antibodies against Influenza A(H7N9) Virus

Qiu, Chao; Huang, Yang; Zhang, Anli; Tian, Di; Wan, Yanmin; Zhang, Wanju; Zhang, Zhiyong; Yuan, Zhenghong; Hu, Yunwen; Zhang, Xiaoyan; Xu, Jianqing

doi:10.3201/eid1910.130728

Cited by 41 publications

(57 citation statements)

References 5 publications

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“…We have previously demonstrated that the titres quantified by Nab assay correlated well with the titres measured by traditional HI assay, using serum samples from influenza A(H7N9)-infected patients and uninfected subjects with good correlation (Spearman r=0.88) [10]. The pseudoviruses were prepared as described in our previous report [10].…”

Section: Pseudovirus-based Neutralisation Assaymentioning

confidence: 95%

“…Previous H7 subtype influenza virus infections in humans such as the influenza A(H7N7) outbreak in Netherlands in 2003 were poorly immunogenic and serodiagnosis and seroepidemiology were challenging [9]. We explored the kinetics of the serological responses to this novel virus in haemagglutination inhibition (HI) assays and in a recently developed H7N9 pseudotype virus particle neutralisation (Nab) test [10]. Viral pseudotypes have been previously shown to provide reliable correlation with conventional microneutralisation tests for influenza A(H5N1) serological studies [11].…”

Section: Introductionmentioning

confidence: 99%

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Kinetics of serological responses in influenza A(H7N9)-infected patients correlate with clinical outcome in China, 2013

et al. 2013

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Section: Pseudovirus-based Neutralisation Assaymentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Kinetics of serological responses in influenza A(H7N9)-infected patients correlate with clinical outcome in China, 2013

et al. 2013

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“…Recombinant H7N9 pseudovirus was produced as previously described (23) with modification. In brief, 10 7 293T cells were co-transfected with lentivirus vector pNL4-3-Luc R-E- (10 μg DNA), pVKD-HA (5 μg DNA) and pVKD-NA (5 μg DNA).…”

Section: Methodsmentioning

confidence: 99%

Coated protein nanoclusters from influenza H7N9 HA are highly immunogenic and induce robust protective immunity

Wang

Chang

et al. 2017

Nanomedicine: Nanotechnology, Biology and Medicine

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Recurring influenza viruses pose an annual threat to public health. A time-saving, cost-effective and egg-independent influenza vaccine approach is important particularly when responding to an emerging pandemic. We fabricated coated, two-layer protein nanoclusters from recombinant trimeric hemagglutinin from an avian-origin H7N9 influenza A virus as an approach for vaccine development in response to an emerging pandemic. Assessment of the virus-specific immune responses and protective efficacy in mice immunized with the nanoclusters demonstrated that the vaccine candidates were highly immunogenic, able to induce protective immunity and long-lasting humoral antibody responses to this virus without the use of adjuvants. Because the advantages of the highly immunogenic coated nanoclusters also include rapid productions in an egg-independent system, this approach has great potential for influenza vaccine production not only in response to an emerging pandemic, but also as a replacement for conventional seasonal influenza vaccines.

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“…H7N9 pseudovirus was obtained through cotransfection with plasmids encoding Env-defective, luciferase-expressing HIV-1 genome (pNL4-3.luc.RE), pVKD-H7HA and pVKD-N9NA from A/Shanghai/4664T/2013 (H7N9) (Qiu et al, 2013). Pseudovirus-containing supernatant harvested 72 h posttransfection was incubated with serially diluted mouse sera at 37 uC for 1 h before adding to confluent MDCK cells in 96-well cell culture plates after washing with PBS.…”

Section: Characterization Of Recombinant Proteins By Sds-page Andmentioning

confidence: 99%

Putative suppressing effect of IgG Fc-conjugated haemagglutinin (HA) stalk of influenza virus H7N9 on the neutralizing immunogenicity of Fc-conjugated HA head: implication for rational design of HA-based influenza vaccines

Xia

et al. 2016

Journal of General Virology

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The emergence of influenza A H7N9 in infection has posed a great threat to public health globally. Poor immunogenicity of H7N9 haemagglutinin (HA) is a major obstacle to the development of an effective H7N9 vaccine. Here, we found that the vaccine containing the H7HA head conjugated with IgG Fc (Hd-Fc) induced strong neutralizing antibody responses and protection against H7N9 infection, whilst the Fc-conjugated H7HA stalk (St-Fc)-based vaccine could not induce neutralizing antibodies, although the St-Fc-immunized mice were partially protected. The vaccines containing the full-length extracellular domain of HA conjugated with Fc and the mixture of Hd-Fc plus St-Fc induced significantly lower neutralizing antibody and haemagglutination inhibition titres than the Hd-Fc-based vaccine. These results suggest that the St-Fc may have inhibitory effects on the neutralizing immunogenicity of Hd-Fc. Therefore, the neutralizing domain(s), such as the receptor-binding domain, in the HA head should be kept and the non-neutralizing domain(s) in the HA stalk with the ability to potentially suppress the neutralizing immunogenicity of HA head should be removed from Fc-conjugated HA-based influenza vaccines to increase the neutralizing antibody response.

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Safe Pseudovirus-based Assay for Neutralization Antibodies against Influenza A(H7N9) Virus

Cited by 41 publications

References 5 publications

Kinetics of serological responses in influenza A(H7N9)-infected patients correlate with clinical outcome in China, 2013

Kinetics of serological responses in influenza A(H7N9)-infected patients correlate with clinical outcome in China, 2013

Coated protein nanoclusters from influenza H7N9 HA are highly immunogenic and induce robust protective immunity

Putative suppressing effect of IgG Fc-conjugated haemagglutinin (HA) stalk of influenza virus H7N9 on the neutralizing immunogenicity of Fc-conjugated HA head: implication for rational design of HA-based influenza vaccines

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