The Experts below are selected from a list of 24486 Experts worldwide ranked by ideXlab platform
John K. Rose - One of the best experts on this subject based on the ideXlab platform.
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Assessment of influenza Virus Hemagglutinin stalk-based immunity in ferrets
Journal of Virology, 2014Co-Authors: Florian Krammer, Rong Hai, Mark A. Yondola, Gene S. Tan, Victor H. Leyva-grado, Alex B. Ryder, Matthew S. Miller, John K. Rose, Peter Palese, Adolfo García-sastreAbstract:ABSTRACT Therapeutic monoclonal antibodies that target the conserved stalk domain of the influenza Virus Hemagglutinin and stalk-based universal influenza Virus vaccine strategies are being developed as promising countermeasures for influenza Virus infections. The pan-H1-reactive monoclonal antibody 6F12 has been extensively characterized and shows broad efficacy against divergent H1N1 strains in the mouse model. Here we demonstrate its efficacy against a pandemic H1N1 challenge Virus in the ferret model of influenza disease. Furthermore, we recently developed a universal influenza Virus vaccine strategy based on chimeric Hemagglutinin constructs that focuses the immune response on the conserved stalk domain of the Hemagglutinin. Here we set out to test this vaccination strategy in the ferret model. Both strategies, pretreatment of animals with a stalk-reactive monoclonal antibody and vaccination with chimeric Hemagglutinin-based constructs, were able to significantly reduce viral titers in nasal turbinates, lungs, and olfactory bulbs. In addition, vaccinated animals also showed reduced nasal wash viral titers. In summary, both strategies showed efficacy in reducing viral loads after an influenza Virus challenge in the ferret model. IMPORTANCE Influenza Virus Hemagglutinin stalk-reactive antibodies tend to be less potent yet are more broadly reactive and can neutralize seasonal and pandemic influenza Virus strains. The ferret model was used to assess the potential of Hemagglutinin stalk-based immunity to provide protection against influenza Virus infection. The novelty and significance of the findings described in this report support the development of vaccines stimulating stalk-specific antibody responses.
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vaccination with a recombinant vesicular stomatitis Virus expressing an influenza Virus Hemagglutinin provides complete protection from influenza Virus challenge
Journal of Virology, 1998Co-Authors: Anjeanette Roberts, Evelyne Kretzschmar, Archibald S Perkins, John P Forman, Ryan Price, Linda Buonocore, Yoshihiro Kawaoka, John K. RoseAbstract:Since the development of a system for generating vesicular stomatitis Virus (VSV) from plasmid DNAs, our laboratory has reported the expression of several different glycoproteins from recombinant VSVs. In one of these studies, high-level expression of an influenza Virus Hemagglutinin (HA) from a recombinant VSV-HA and efficient incorporation of the HA protein into the virions was reported (E. Kretzschmar, L. Buonocore, M. J. Schnell, and J. K. Rose, J. Virol. 71:5982–5989, 1997). We report here that VSV-HA is an effective intranasal vaccine vector that raises high levels of neutralizing antibody to influenza Virus and completely protects mice from bronchial pneumonia caused by challenge with a lethal dose of influenza A Virus. Additionally, these recombinant VSVs are less pathogenic than wild-type VSV (serotype Indiana). This vector-associated pathogenicity was subsequently eliminated through introduction of specific attenuating deletions. These live attenuated recombinant VSVs have great potential as vaccine vectors.
Thomas D Madden - One of the best experts on this subject based on the ideXlab platform.
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nucleoside modified mrna immunization elicits influenza Virus Hemagglutinin stalk specific antibodies
Nature Communications, 2018Co-Authors: Norbert Pardi, Kaela Parkhouse, Ericka Kirkpatrick, Meagan Mcmahon, Seth J Zost, Barbara L Mui, Ying K Tam, Katalin Kariko, Christopher J Barbosa, Thomas D MaddenAbstract:Currently available influenza Virus vaccines have inadequate effectiveness and are reformulated annually due to viral antigenic drift. Thus, development of a vaccine that confers long-term protective immunity against antigenically distant influenza Virus strains is urgently needed. The highly conserved influenza Virus Hemagglutinin (HA) stalk represents one of the potential targets of broadly protective/universal influenza Virus vaccines. Here, we evaluate a potent broadly protective influenza Virus vaccine candidate that uses nucleoside-modified and purified mRNA encoding full-length influenza Virus HA formulated in lipid nanoparticles (LNPs). We demonstrate that immunization with HA mRNA-LNPs induces antibody responses against the HA stalk domain of influenza Virus in mice, rabbits, and ferrets. The HA stalk-specific antibody response is associated with protection from homologous, heterologous, and heterosubtypic influenza Virus infection in mice.
Wolfgang Garten - One of the best experts on this subject based on the ideXlab platform.
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Cleavage of influenza Virus Hemagglutinin by host cell proteases
International Congress Series, 2004Co-Authors: Wolfgang Garten, Mikhail Matrosovich, Tatyana Dr. Matrosovich, Markus Eickmann, A VahhabzadehAbstract:Abstract A full-length gene of tryptase TMPRSS2 was cloned from total RNA of human tracheo-bronchial epithelial (HTBE) cells. The TMRSS2 protein was expressed in Escherichia coli as two truncated forms. Recombinant TMPRSS2 was enzymatically active, cleaved influenza A Virus Hemagglutinin and activated multi-cycle Virus replication in MDCK cells. Thus, TMPRSS2 is an appropriate tryptase candidate for the HA-activation of influenza A Viruses in vivo.
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influenza Virus Hemagglutinin with multibasic cleavage site is activated by furin a subtilisin like endoprotease
The EMBO Journal, 1992Co-Authors: A Stienekegrober, Hans-dieter Klenk, Herbert Angliker, E Shaw, G Thomas, C Roberts, Wolfgang GartenAbstract:: Many Viruses have membrane glycoproteins that are activated at cleavage sites containing multiple arginine and lysine residues by cellular proteases so far not identified. The proteases responsible for cleavage of the Hemagglutinin of fowl plague Virus, a prototype of these glycoproteins, has now been isolated from Madin-Darby bovine kidney cells. The enzyme has a mol. wt of 85,000, a pH optimum ranging from 6.5 to 7.5, is calcium dependent and recognizes the consensus sequence R-X-K/R-R at the cleavage site of the Hemagglutinin. Using a specific antiserum it has been identified as furin, a subtilisin-like eukaryotic protease. The fowl plague Virus Hemagglutinin was also cleaved after coexpression with human furin from cDNA by vaccinia Virus vectors. Peptidyl chloroalkylketones containing the R-X-K/R-R motif specifically bind to the catalytic site of furin and are therefore potent inhibitors of Hemagglutinin cleavage and fusion activity.
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Modification of the cleavage activation of the influenza Virus Hemagglutinin by site-specific mutagenesis.
Behring Institute Mitteilungen, 1991Co-Authors: Wolfgang Garten, M. Vey, Ohuchi R, Masanobu Ohuchi, Hans-dieter KlenkAbstract:Factors determining cleavability of influenza Virus Hemagglutinin which is activated by ubiquitous cellular endoproteases were analysed by carrying out site-directed mutagenesis on the cloned hemaglutinin genes of strains A/FPV/Rostock/34 (subtype H7) and A/Port Chalmers/1/73 (subtype H3). Substitutions at the cleavage site of the H7 Hemagglutinin indicate that the tetrapeptide Arg-X-Lys/Arg-Arg is the minimal consensus sequence recognized by the ubiquitous proteases. The H3 Hemagglutinin also became susceptible to these enzymes, when additional arginines were inserted at the cleavage site. Three arginines were sufficient, when the carbohydrate was removed, whereas four additional arginines are needed when this carbohydrate was present, indicating that the accessibility of the cleavage motif is important for the protease. The appropriate localization of the basic cleavage motif within the amino acid sequence and the spatial structure of the Hemagglutinin precursor is an additional prerequisite for cleavage.
Florian Krammer - One of the best experts on this subject based on the ideXlab platform.
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broadly cross reactive nonneutralizing antibodies against influenza b Virus Hemagglutinin demonstrate effector function dependent protection against lethal viral challenge in mice
Journal of Virology, 2019Co-Authors: Guha Asthagiri Arunkumar, Andriani Ioannou, Teddy John Wohlbold, Philip Meade, Sadaf Aslam, Fatima Amanat, Juan Ayllon, Adolfo Garciasastre, Florian KrammerAbstract:Protection from influenza Virus infection is canonically associated with antibodies that neutralize the Virus by blocking the interaction between the viral Hemagglutinin and host cell receptors. However, protection can also be conferred by other mechanisms, including antibody-mediated effector functions. Here, we report the characterization of 22 broadly cross-reactive, nonneutralizing antibodies specific for influenza B Virus Hemagglutinin. The majority of these antibodies recognized influenza B Viruses isolated over the period of 73 years and bind the conserved stalk domain of the Hemagglutinin. A proportion of the characterized antibodies protected mice from both morbidity and mortality after challenge with a lethal dose of influenza B Virus. Activity in an antibody-dependent cell-mediated cytotoxicity reporter assay correlated strongly with protection, suggesting that Fc-dependent effector function determines protective efficacy. The information regarding mechanism of action and epitope location stemming from our characterization of these antibodies will inform the design of urgently needed vaccines that could induce broad protection against influenza B Viruses. IMPORTANCE While broadly protective antibodies against the influenza A Virus Hemagglutinin have been well studied, very limited information is available for antibodies that broadly recognize influenza B Viruses. Similarly, the development of a universal or broadly protective influenza B Virus vaccine lags behind the development of such a vaccine for influenza A Virus. More information about epitope location and mechanism of action of broadly protective influenza B Virus antibodies is required to inform vaccine development. In addition, protective antibodies could be a useful tool to treat or prevent influenza B Virus infection in pediatric cohorts or in a therapeutic setting in immunocompromised individuals in conjugation with existing treatment avenues.
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Antibody Responses toward the Major Antigenic Sites of Influenza B Virus Hemagglutinin in Mice, Ferrets, and Humans.
Journal of Virology, 2018Co-Authors: Weina Sun, Florian Krammer, Davina S. Kang, Allen Zheng, Sean T. H. Liu, Felix Broecker, Viviana Simon, Peter PaleseAbstract:ABSTRACT The influenza B Virus Hemagglutinin contains four major antigenic sites (the 120 loop, the 150 loop, the 160 loop, and the 190 helix) within the head domain. These immunodominant antigenic sites are the main targets of neutralizing antibodies and are subject to antigenic drift. Yet little is known about the specific antibody responses toward each site in terms of antibody prevalence and hemagglutination inhibition activity. In this study, we used modified Hemagglutinins of influenza B Virus which display only one or none of the major antigenic sites to measure antibody responses toward the classical as well as the noncanonical epitopes in mice, ferrets, and humans. With our novel reagents, we found that both hemagglutination inhibition antibodies and total IgGs were mostly induced by the major antigenic sites. However, in human adults, we observed high hemagglutination inhibition antibody responses toward the noncanonical epitopes. By stratifying the human samples into age groups, we found that the noncanonical antibody responses appeared to increase with age. IMPORTANCE This study dissected the specific antibody responses toward the major antigenic sites and the noncanonical epitopes of influenza B Virus Hemagglutinin in animals and humans using novel reagents. These findings will guide the design of the next generation of influenza Virus vaccines.
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Assessment of influenza Virus Hemagglutinin stalk-based immunity in ferrets
Journal of Virology, 2014Co-Authors: Florian Krammer, Rong Hai, Mark A. Yondola, Gene S. Tan, Victor H. Leyva-grado, Alex B. Ryder, Matthew S. Miller, John K. Rose, Peter Palese, Adolfo García-sastreAbstract:ABSTRACT Therapeutic monoclonal antibodies that target the conserved stalk domain of the influenza Virus Hemagglutinin and stalk-based universal influenza Virus vaccine strategies are being developed as promising countermeasures for influenza Virus infections. The pan-H1-reactive monoclonal antibody 6F12 has been extensively characterized and shows broad efficacy against divergent H1N1 strains in the mouse model. Here we demonstrate its efficacy against a pandemic H1N1 challenge Virus in the ferret model of influenza disease. Furthermore, we recently developed a universal influenza Virus vaccine strategy based on chimeric Hemagglutinin constructs that focuses the immune response on the conserved stalk domain of the Hemagglutinin. Here we set out to test this vaccination strategy in the ferret model. Both strategies, pretreatment of animals with a stalk-reactive monoclonal antibody and vaccination with chimeric Hemagglutinin-based constructs, were able to significantly reduce viral titers in nasal turbinates, lungs, and olfactory bulbs. In addition, vaccinated animals also showed reduced nasal wash viral titers. In summary, both strategies showed efficacy in reducing viral loads after an influenza Virus challenge in the ferret model. IMPORTANCE Influenza Virus Hemagglutinin stalk-reactive antibodies tend to be less potent yet are more broadly reactive and can neutralize seasonal and pandemic influenza Virus strains. The ferret model was used to assess the potential of Hemagglutinin stalk-based immunity to provide protection against influenza Virus infection. The novelty and significance of the findings described in this report support the development of vaccines stimulating stalk-specific antibody responses.
Peter Palese - One of the best experts on this subject based on the ideXlab platform.
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Antibody Responses toward the Major Antigenic Sites of Influenza B Virus Hemagglutinin in Mice, Ferrets, and Humans.
Journal of Virology, 2018Co-Authors: Weina Sun, Florian Krammer, Davina S. Kang, Allen Zheng, Sean T. H. Liu, Felix Broecker, Viviana Simon, Peter PaleseAbstract:ABSTRACT The influenza B Virus Hemagglutinin contains four major antigenic sites (the 120 loop, the 150 loop, the 160 loop, and the 190 helix) within the head domain. These immunodominant antigenic sites are the main targets of neutralizing antibodies and are subject to antigenic drift. Yet little is known about the specific antibody responses toward each site in terms of antibody prevalence and hemagglutination inhibition activity. In this study, we used modified Hemagglutinins of influenza B Virus which display only one or none of the major antigenic sites to measure antibody responses toward the classical as well as the noncanonical epitopes in mice, ferrets, and humans. With our novel reagents, we found that both hemagglutination inhibition antibodies and total IgGs were mostly induced by the major antigenic sites. However, in human adults, we observed high hemagglutination inhibition antibody responses toward the noncanonical epitopes. By stratifying the human samples into age groups, we found that the noncanonical antibody responses appeared to increase with age. IMPORTANCE This study dissected the specific antibody responses toward the major antigenic sites and the noncanonical epitopes of influenza B Virus Hemagglutinin in animals and humans using novel reagents. These findings will guide the design of the next generation of influenza Virus vaccines.
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Assessment of influenza Virus Hemagglutinin stalk-based immunity in ferrets
Journal of Virology, 2014Co-Authors: Florian Krammer, Rong Hai, Mark A. Yondola, Gene S. Tan, Victor H. Leyva-grado, Alex B. Ryder, Matthew S. Miller, John K. Rose, Peter Palese, Adolfo García-sastreAbstract:ABSTRACT Therapeutic monoclonal antibodies that target the conserved stalk domain of the influenza Virus Hemagglutinin and stalk-based universal influenza Virus vaccine strategies are being developed as promising countermeasures for influenza Virus infections. The pan-H1-reactive monoclonal antibody 6F12 has been extensively characterized and shows broad efficacy against divergent H1N1 strains in the mouse model. Here we demonstrate its efficacy against a pandemic H1N1 challenge Virus in the ferret model of influenza disease. Furthermore, we recently developed a universal influenza Virus vaccine strategy based on chimeric Hemagglutinin constructs that focuses the immune response on the conserved stalk domain of the Hemagglutinin. Here we set out to test this vaccination strategy in the ferret model. Both strategies, pretreatment of animals with a stalk-reactive monoclonal antibody and vaccination with chimeric Hemagglutinin-based constructs, were able to significantly reduce viral titers in nasal turbinates, lungs, and olfactory bulbs. In addition, vaccinated animals also showed reduced nasal wash viral titers. In summary, both strategies showed efficacy in reducing viral loads after an influenza Virus challenge in the ferret model. IMPORTANCE Influenza Virus Hemagglutinin stalk-reactive antibodies tend to be less potent yet are more broadly reactive and can neutralize seasonal and pandemic influenza Virus strains. The ferret model was used to assess the potential of Hemagglutinin stalk-based immunity to provide protection against influenza Virus infection. The novelty and significance of the findings described in this report support the development of vaccines stimulating stalk-specific antibody responses.
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guiding the immune response against influenza Virus Hemagglutinin toward the conserved stalk domain by hyperglycosylation of the globular head domain
Journal of Virology, 2014Co-Authors: Dirk Eggink, Peter H Goff, Peter PaleseAbstract:ABSTRACT Influenza Virus Hemagglutinin consists of a highly variable and immunodominant head domain and a more conserved but immunosubdominant stalk domain. We introduced seven N-linked glycosylation sites in the Hemagglutinin head domain to shield the immunodominant antigenic sites. The hyperglycosylated Hemagglutinin enhanced stalk-directed seroreactivity while dampening the head response in immunized mice. Upon influenza Virus challenge, mice vaccinated with the hyperglycosylated Hemagglutinin were better protected against morbidity and mortality than mice receiving the wild-type Hemagglutinin.
