Broadly Inhibiting Antineuraminidase Monoclonal Antibodies Induced by Trivalent Influenza Vaccine and H7N9 Infection in Humans

ABSTRACT The majority of antibodies induced by influenza neuraminidase (NA), like those against hemagglutinin (HA), are relatively specific to viruses isolated within a limited time window, as seen in serological studies and the analysis of many murine monoclonal antibodies (MAbs). We report three broadly reactive human MAbs targeting N1 NA. Two were isolated from a young adult vaccinated with trivalent influenza vaccine (TIV), which inhibited N1 NA from viruses isolated from humans over a period of a hundred years. The third antibody, isolated from a child with acute mild H7N9 infection, inhibited both group 1 N1 and group 2 N9 NAs. In addition, the antibodies cross-inhibited the N1 NAs of highly pathogenic avian H5N1 influenza viruses. These antibodies are protective in prophylaxis against seasonal H1N1 viruses in mice. This study demonstrates that human antibodies to N1 NA with exceptional cross-reactivity can be recalled by vaccination and highlights the importance of standardizing the NA antigen in seasonal vaccines to offer optimal protection. IMPORTANCE Antibodies to the influenza virus NA can provide protection against influenza disease. Analysis of human antibodies to NA lags behind that of antibodies to HA. We show that human monoclonal antibodies against NA induced by vaccination and infection can be very broadly reactive, with the ability to inhibit a wide spectrum of N1 NAs on viruses isolated between 1918 and 2018. This suggests that antibodies to NA may be a useful therapy and that the efficacy of influenza vaccines could be enhanced by ensuring the appropriate content of NA antigen.

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Broadly inhibiting anti-neuraminidase monoclonal antibodies induced by trivalent influenza vaccine and H7N9 infection in humans

10.1101/682450 ◽

2019 ◽

Cited By ~ 1

Author(s):

Pramila Rijal ◽

Bei Bei Wang ◽

Tiong Kit Tan ◽

Lisa Schimanski ◽

Philipp Janesch ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Influenza Vaccine ◽

Influenza Viruses ◽

List Type ◽

Human Monoclonal Antibodies ◽

Human Antibodies ◽

Trivalent Influenza Vaccine ◽

H7n9 Infection ◽

Group 2 ◽

Group 1

AbstractThe majority of antibodies induced by influenza neuraminidase (NA), like those against hemagglutinin (HA), are relatively specific to viruses isolated within a limited time-window as seen in serological studies and the analysis of many murine monoclonal antibodies. We report three broadly reactive human monoclonal antibodies (mAbs) targeting N1 NA. Two were isolated from a young adult vaccinated with trivalent influenza vaccine (TIV), which inhibited N1 NA from viruses isolated from human over a period of a hundred years. The third antibody isolated from a child with acute mild H7N9 infection inhibited both group 1 N1 and group 2 N9 NAs. In addition, the antibodies cross-inhibited the N1 NAs of highly pathogenic avian H5N1 influenza viruses. These antibodies are protective in prophylaxis against seasonal H1N1 viruses in mice. This study demonstrates that human antibodies to N1 NA with exceptional cross-reactivity can be recalled by vaccination and highlights the importance of standardizing the NA antigen in seasonal vaccines to offer optimal protection.ImportanceAntibodies to the influenza NA can provide protection against influenza disease. Analysis of human antibodies to NA lags behind that for HA. We show that human monoclonal antibodies against NA induced by vaccination and infection can be very broadly reactive and able to inhibit a wide spectrum of N1 NAs between 1918 and 2018. This suggests that antibodies to NA may be a useful therapy, and that efficacy of influenza vaccines could be enhanced by ensuring appropriate content of NA antigen.Highlights of the paperAntibodies that inhibit influenza viruses with N1 neuraminidase (NA), with broad reactivity for viruses isolated between 1918-2018, can be isolated from human recipients of seasonal influenza vaccineAntibodies targeting N1 NA of human seasonal H1N1 viruses can cross-react with a variety of avian N1 neuraminidasesAcute H7N9 infection can recall memory B cells to N1 NA and elicit cross-reactive antibodies to the group 1 N1 and group 2 N9 NAsAntibodies to N1 NA with this broad reactivity protect against lethal virus challenge

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Properties and Therapeutic Efficacy of Broadly Reactive Chimeric and Humanized H5-Specific Monoclonal Antibodies against H5N1 Influenza Viruses

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01436-10 ◽

2011 ◽

Vol 55 (4) ◽

pp. 1349-1357 ◽

Cited By ~ 9

Author(s):

Qingbing Zheng ◽

Lin Xia ◽

Wai Lan Wu ◽

Zhenhua Zheng ◽

Yongting Huo ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Virus Infection ◽

H5n1 Virus ◽

Influenza Viruses ◽

Chimeric Antibody ◽

Dependent Manner ◽

Variable Regions ◽

H5n1 Influenza ◽

Therapeutic Regime ◽

Humanized Antibodies

ABSTRACTHighly pathogenic H5N1 virus infection causes severe disease and a high rate of fatality in humans. Development of humanized monoclonal antibodies may provide an efficient therapeutic regime for H5N1 virus infection. In the present study, broadly cross-reactive monoclonal antibodies (MAbs) derived from mice were humanized to minimize immunogenicity. One chimeric antibody (cAb) and seven humanized antibodies (hAbs) were constructed. These antibodies retained broad-spectrum reactivity to H5N1 viruses, binding to recombinant H5-subtype HA1 molecules expressed in CHO cells in a dose-dependent manner and exhibiting similar reactivities against antigenically distinct H5N1 viruses in hemagglutination inhibition (HI) assays. One humanized antibody, 37 hAb, showed HI and neutralization activities comparable to that of the parental murine antibody, 13D4 MAb, while the other six antibodies were less reactive to H5N1 viruses. Analysis of amino acid sequences in the variable region frameworks of the seven humanized antibodies found that Q5 and Y27 in the VH region are highly conserved murine residues. Comparison of the three-dimensional structures derived from the variable regions of MAbs 37 hAb, H1202-34, and 13D4 revealed that residue substitutions at sites 70 and 46 may be the major cause for the observed differences in binding affinity. Examination of the chimeric antibody and one of the humanized antibodies, 37 hAb, showed that both antibodies offered postinfection protection against lethal challenge with antigenically diverse H5N1 viruses in the mouse model. Chimeric and humanized antibodies which retain the broadly reactive and protective properties of murine H5-specific monoclonal antibodies have great potential for use in the treatment of human H5N1 infection.

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Human antibodies that neutralize respiratory droplet transmissible H5N1 influenza viruses

Journal of Clinical Investigation ◽

10.1172/jci69377 ◽

2013 ◽

Vol 123 (10) ◽

pp. 4405-4409 ◽

Cited By ~ 24

Author(s):

Natalie J. Thornburg ◽

David P. Nannemann ◽

David L. Blum ◽

Jessica A. Belser ◽

Terrence M. Tumpey ◽

...

Keyword(s):

Influenza Viruses ◽

Human Antibodies ◽

H5n1 Influenza

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Antibodies directed towards neuraminidase N1 control disease in a mouse model of influenza

Journal of Virology ◽

10.1128/jvi.01584-17 ◽

2017 ◽

pp. JVI.01584-17 ◽

Cited By ~ 4

Author(s):

E.R. Job ◽

M. Schotsaert ◽

L.I. Ibañez ◽

A. Smet ◽

T. Ysenbaert ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Antiviral Activity ◽

Influenza A ◽

Conformational Epitope ◽

Influenza Viruses ◽

Lethal Challenge ◽

H5n1 Influenza ◽

Antigenic Properties

There is increasing evidence to suggest that antibodies directed towards influenza A virus (IAV) neuraminidase (NA) are an important correlate of protection against influenza in humans. Moreover, the potential of NA-specific antibodies to provide broader protection than conventional hemagglutinin (HA) antibodies has been recognized. Herein, we describe the isolation of two monoclonal antibodies, N1-7D3 and N1-C4, directed towards the N1 NA. N1-7D3 binds to a conserved linear epitope in the membrane distal, carboxy-terminal part of the NA and reacted with the NA of seasonal H1N1 isolates ranging from 1977 till 2007 the 2009 H1N1pdm virus as well as A/Vietnam/1194/04 (H5N1). However, N1-7D3 lacked NA inhibition (NI) activity and the ability to protect BALB/c mice against a lethal challenge with a range of H1N1 viruses. Conversely, N1-C4 bound to a conformational epitope that is conserved between two influenza subtypes, the 2009 H1N1pdm and H5N1 IAV and displayed potentin vitroantiviral activity mediating both NI and plaque size-reduction. Moreover, N1-C4 could provide heterosubtypic protection in BALB/c mice against a lethal challenge with 2009 H1N1pdm or H5N1 virus. Glutamic acid residue 311 in the NA was found to be critical for the NA binding and antiviral activity of monoclonal antibody N1-C4. Our data provide further evidence on cross-protective epitopes within the N1 subtype and highlight the potential of NA as an important target for vaccine and therapeutic approaches.ImportanceInfluenza remains a world-wide burden to public health. As such the development of new and novel vaccines and therapeutics against influenza virus is crucial. Human challenge studies have recently highlighted the importance of antibodies directed towards the viral neuraminidase (NA) as an important correlate of reduced influenza-associated disease severity. Furthermore, there is evidence that anti-NA antibodies can provide broader protection than antibodies towards the viral hemagglutinin. Here we describe the isolation and detailed characterization of two N1 NA-specific monoclonal antibodies. One of these monoclonal antibodies broadly binds N1 type NAs and the second one displays NAI, in vitro and in vivo anti-viral activity against 2009 H1N1pdm and H5N1 influenza viruses. These two new anti-NA antibodies contribute to our understanding of the antigenic properties and protective potential of the influenza NA antigen.

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Transgenic Chicks Expressing Interferon-Inducible Transmembrane Protein 1 (IFITM1) Restrict Highly Pathogenic H5N1 Influenza Viruses

International Journal of Molecular Sciences ◽

10.3390/ijms22168456 ◽

2021 ◽

Vol 22 (16) ◽

pp. 8456

Author(s):

Mohammed A. Rohaim ◽

Mohammad Q. Al-Natour ◽

Mohammed A. Abdelsabour ◽

Rania F. El Naggar ◽

Yahia M. Madbouly ◽

...

Keyword(s):

Influenza Virus ◽

Mammalian Cells ◽

Transmembrane Protein ◽

Wide Spectrum ◽

Lethal Dose ◽

Influenza Viruses ◽

Highly Pathogenic ◽

Zoonotic Infections ◽

H5n1 Influenza ◽

Transgenic Chickens

Mammalian cells utilize a wide spectrum of pathways to antagonize the viral replication. These pathways are typically regulated by antiviral proteins and can be constitutively expressed but also exacerbated by interferon induction. A myriad of interferon-stimulated genes (ISGs) have been identified in mounting broad-spectrum antiviral responses. Members of the interferon-induced transmembrane (IFITM) family of proteins are unique among these ISGs due to their ability to prevent virus entry through the lipid bilayer into the cell. In the current study, we generated transgenic chickens that constitutively and stably expressed chicken IFITM1 (chIFITM1) using the avian sarcoma-leukosis virus (RCAS)-based gene transfer system. The challenged transgenic chicks with clinical dose 104 egg infective dose 50 (EID50) of highly pathogenic avian influenza virus (HPAIV) subtype H5N1 (clade 2.2.1.2) showed 100% protection and significant infection tolerance. Although challenged transgenic chicks displayed 60% protection against challenge with the sub-lethal dose (EID50 105), the transgenic chicks showed delayed clinical symptoms, reduced virus shedding, and reduced histopathologic alterations compared to non-transgenic challenged control chickens. These finding indicate that the sterile defense against H5N1 HPAIV offered by the stable expression of chIFITM1 is inadequate; however, the clinical outcome can be substantially ameliorated. In conclusion, chIFITM proteins can inhibit influenza virus replication that can infect various host species and could be a crucial barrier against zoonotic infections.

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Design and Characterization of a Computationally Optimized Broadly Reactive Hemagglutinin Vaccine for H1N1 Influenza Viruses

Journal of Virology ◽

10.1128/jvi.03152-15 ◽

2016 ◽

Vol 90 (9) ◽

pp. 4720-4734 ◽

Cited By ~ 72

Author(s):

Donald M. Carter ◽

Christopher A. Darby ◽

Bradford C. Lefoley ◽

Corey J. Crevar ◽

Timothy Alefantis ◽

...

Keyword(s):

Influenza Vaccine ◽

Hemagglutination Inhibition ◽

H1n1 Influenza ◽

Influenza Viruses ◽

Pandemic H1n1 ◽

Vaccine Strategy ◽

First Report ◽

The Past ◽

H5n1 Influenza

ABSTRACTOne of the challenges of developing influenza A vaccines is the diversity of antigenically distinct isolates. Previously, a novel hemagglutinin (HA) for H5N1 influenza was derived from a methodology termed computationally optimized broadly reactive antigen (COBRA). This COBRA HA elicited a broad antibody response against H5N1 isolates from different clades. We now report the development and characterization of a COBRA-based vaccine for both seasonal and pandemic H1N1 influenza virus isolates. Nine prototype H1N1 COBRA HA proteins were developed and tested in mice using a virus-like particle (VLP) format for the elicitation of broadly reactive, functional antibody responses and protection against viral challenge. These candidates were designed to recognize H1N1 viruses isolated within the last 30 years. In addition, several COBRA candidates were designed based on sequences of H1N1 viruses spanning the past 100 years, including modern pandemic H1N1 isolates. Four of the 9 H1N1 COBRA HA proteins (X1, X3, X6, and P1) had the broadest hemagglutination inhibition (HAI) activity against a panel of 17 H1N1 viruses. These vaccines were used in cocktails or prime-boost combinations. The most effective regimens that both elicited the broadest HAI response and protected mice against a pandemic H1N1 challenge were vaccines that contained the P1 COBRA VLP and either the X3 or X6 COBRA VLP vaccine. These mice had little or no detectable viral replication, comparable to that observed with a matched licensed vaccine. This is the first report describing a COBRA-based HA vaccine strategy that elicits a universal, broadly reactive, protective response against seasonal and pandemic H1N1 isolates.IMPORTANCEUniversal influenza vaccine approaches have the potential to be paradigm shifting for the influenza vaccine field, with the goal of replacing the current standard of care with broadly cross-protective vaccines. We have used COBRA technology to develop an HA head-based strategy that elicits antibodies against many H1 strains that have undergone genetic drift and has potential as a “subtype universal” vaccine. Nine HA COBRA candidates were developed, and these vaccines were used alone, in cocktails or in prime-boost combinations. The most effective regimens elicited the broadest hemagglutination inhibition (HAI) response against a panel of H1N1 viruses isolated over the past 100 years. This is the first report describing a COBRA-based HA vaccine strategy that elicits a broadly reactive response against seasonal and pandemic H1N1 isolates.

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PECULARITIES OF DEVELOPMENT OF REASSORTANT STRAINS FOR LIVE INFLUENZA VACCINE BASED ON VIRUSES WITH PANDEMIC POTENTIAL

Medical academic journal ◽

10.17816/maj18254-62 ◽

2018 ◽

Vol 18 (2) ◽

pp. 54-62

Author(s):

N V Larionova ◽

I V Kiseleva ◽

E M Doroshenko ◽

L G Rudenko

Keyword(s):

Avian Influenza ◽

Influenza Vaccine ◽

Pandemic Influenza ◽

Influenza A ◽

Swine Influenza ◽

Influenza Viruses ◽

Genome Composition ◽

Live Attenuated Influenza Vaccine ◽

Ha Gene ◽

H5n1 Influenza

Objective of the research: to characterize the effectiveness of obtaining reassortant strains for live influenza vaccine based on pandemic and potentially pandemic influenza viruses which are new for humans. Materials and methods of the research: influenza virus A(H1N1)pdm09, potentially pandemic A(H3N2)v and A(H5N1) influenza viruses, a master donor virus for the Russian live influenza vaccine. Virological and molecular genetics methods of research. Results: strains with a 6:2 genome composition for live influenza reassortant vaccine based on the pandemic '(H1N1)pdm09 virus and swine influenza A(H3N2)v virus have been successfully obtained. The reassortment of highly pathogenic avian influenza (HPAI) viruses A(H5N1) and human A(H2N2) master donor virus for live attenuated influenza vaccine is complicated by the features of the constellation of their genes. H5N2 reassortants which inherited only HA gene from HPAI viruses of avian influenza were obtained. Despite the impossibility of development of 6:2 reassortants, the phenotypic, preclinical characteristics of the reassortants with the 7:1 genome composition, and their further clinical studies on volunteers have shown that such vaccine strains can be successfully used to prevent diseases caused by avian influenza viruses. Conclusion. When vaccine strains against potentially pandemic influenza viruses are obtained by the classical reassortment technique then the success depends on the mutual constellation of genes of phylogenetically distant far apart parental viruses. In some cases the inheritance by the reassortant of a single HA gene from the antigenic actual virus can be a compromise for LAIV development.

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Avian influenza pandemic preparedness: developing prepandemic and pandemic vaccines against a moving target

Expert Reviews in Molecular Medicine ◽

10.1017/s1462399410001432 ◽

2010 ◽

Vol 12 ◽

Cited By ~ 17

Author(s):

Neetu Singh ◽

Aseem Pandey ◽

Suresh K. Mittal

Keyword(s):

Avian Influenza ◽

Influenza Vaccine ◽

Pandemic Influenza ◽

Influenza Pandemic ◽

Intervention Strategies ◽

Influenza Viruses ◽

Current Status ◽

Pandemic Preparedness ◽

H5n1 Influenza ◽

Pandemic Threat

The unprecedented global spread of highly pathogenic avian H5N1 influenza viruses within the past ten years and their extreme lethality to poultry and humans has underscored their potential to cause an influenza pandemic. Combating the threat of an impending H5N1 influenza pandemic will require a combination of pharmaceutical and nonpharmaceutical intervention strategies. The emergence of the H1N1 pandemic in 2009 emphasised the unpredictable nature of a pandemic influenza. Undoubtedly, vaccines offer the most viable means to combat a pandemic threat. Current egg-based influenza vaccine manufacturing strategies are unlikely to be able to cater to the huge, rapid global demand because of the anticipated scarcity of embryonated eggs in an avian influenza pandemic and other factors associated with the vaccine production process. Therefore, alternative, egg-independent vaccine manufacturing strategies should be evaluated to supplement the traditional egg-derived influenza vaccine manufacturing. Furthermore, evaluation of dose-sparing strategies that offer protection with a reduced antigen dose will be critical for pandemic influenza preparedness. Development of new antiviral therapeutics and other, nonpharmaceutical intervention strategies will further supplement pandemic preparedness. This review highlights the current status of egg-dependent and egg-independent strategies against an avian influenza pandemic.

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