The Experts below are selected from a list of 19938 Experts worldwide ranked by ideXlab platform
Olli Vapalahti - One of the best experts on this subject based on the ideXlab platform.
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vaccinia virus free rescue of fluorescent replication defective vesicular stomatitis virus and pseudotyping with puumala virus glycoproteins for use in Neutralization Tests
Journal of General Virology, 2016Co-Authors: Rommel Paneth Iheozorejiofor, Ake Lundkvist, Lev Levanov, Jussi Hepojoki, Tomas Strandin, Alexander Plyusnin, Olli VapalahtiAbstract:Puumala virus (PUUV) grows slowly in cell culture. To study antigenic properties of PUUV, an amenable method for their expression would be beneficial. To achieve this, a replication-defective recombinant vesicular stomatitis virus, rVSVΔG*EGFP, was rescued using BSRT7/5 and encephalomyocarditis virus (EMCV) internal ribosomal entry site (IRES)-enabled rescue plasmids. Using these particles, pseudotypes bearing PUUV Sotkamo strain glycoproteins were produced, with titres in the range 105–108, and were used in pseudotype focus reduction Neutralization Tests (pFRNTs) with neutralizing monoclonal antibodies and patient sera. The results were compared with those from orthodox focus reduction Neutralization Tests (oFRNTs) using native PUUV with the same samples and showed a strong positive correlation (rs = 0.82) between the methods. While developing the system we identified three amino acids which were mutated in the Vero E6 cell culture adapted PUUV prototype Sotkamo strain sequence, and changing these residues was critical for expression and neutralizing antibody binding of PUUV glycoproteins.
Rommel Paneth Iheozorejiofor - One of the best experts on this subject based on the ideXlab platform.
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vaccinia virus free rescue of fluorescent replication defective vesicular stomatitis virus and pseudotyping with puumala virus glycoproteins for use in Neutralization Tests
Journal of General Virology, 2016Co-Authors: Rommel Paneth Iheozorejiofor, Ake Lundkvist, Lev Levanov, Jussi Hepojoki, Tomas Strandin, Alexander Plyusnin, Olli VapalahtiAbstract:Puumala virus (PUUV) grows slowly in cell culture. To study antigenic properties of PUUV, an amenable method for their expression would be beneficial. To achieve this, a replication-defective recombinant vesicular stomatitis virus, rVSVΔG*EGFP, was rescued using BSRT7/5 and encephalomyocarditis virus (EMCV) internal ribosomal entry site (IRES)-enabled rescue plasmids. Using these particles, pseudotypes bearing PUUV Sotkamo strain glycoproteins were produced, with titres in the range 105–108, and were used in pseudotype focus reduction Neutralization Tests (pFRNTs) with neutralizing monoclonal antibodies and patient sera. The results were compared with those from orthodox focus reduction Neutralization Tests (oFRNTs) using native PUUV with the same samples and showed a strong positive correlation (rs = 0.82) between the methods. While developing the system we identified three amino acids which were mutated in the Vero E6 cell culture adapted PUUV prototype Sotkamo strain sequence, and changing these residues was critical for expression and neutralizing antibody binding of PUUV glycoproteins.
Mauro Pistello - One of the best experts on this subject based on the ideXlab platform.
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viral infection Neutralization Tests a focus on severe acute respiratory syndrome coronavirus 2 with implications for convalescent plasma therapy
Reviews in Medical Virology, 2020Co-Authors: Daniele Focosi, Fabrizio Maggi, Paola Mazzetti, Mauro PistelloAbstract:Viral Neutralization Tests (VNTs) have long been considered old-fashioned tricks in the armamentarium of fundamental virology, with laboratory implementation for a limited array of viruses only. Nevertheless, they represent the most reliable surrogate of potency for passive immunotherapies, such as monoclonal or polyclonal antibody therapy. The recent interest around therapy with convalescent plasma or monoclonal antibodies for the Covid-19 pandemic has paralleled the revival of VNTs. We review here the available methods by dissecting variations for each fundamental component of the VNT (i.e., virus type and dose, replication-competent cell line, serum, and detection system).
Ake Lundkvist - One of the best experts on this subject based on the ideXlab platform.
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vaccinia virus free rescue of fluorescent replication defective vesicular stomatitis virus and pseudotyping with puumala virus glycoproteins for use in Neutralization Tests
Journal of General Virology, 2016Co-Authors: Rommel Paneth Iheozorejiofor, Ake Lundkvist, Lev Levanov, Jussi Hepojoki, Tomas Strandin, Alexander Plyusnin, Olli VapalahtiAbstract:Puumala virus (PUUV) grows slowly in cell culture. To study antigenic properties of PUUV, an amenable method for their expression would be beneficial. To achieve this, a replication-defective recombinant vesicular stomatitis virus, rVSVΔG*EGFP, was rescued using BSRT7/5 and encephalomyocarditis virus (EMCV) internal ribosomal entry site (IRES)-enabled rescue plasmids. Using these particles, pseudotypes bearing PUUV Sotkamo strain glycoproteins were produced, with titres in the range 105–108, and were used in pseudotype focus reduction Neutralization Tests (pFRNTs) with neutralizing monoclonal antibodies and patient sera. The results were compared with those from orthodox focus reduction Neutralization Tests (oFRNTs) using native PUUV with the same samples and showed a strong positive correlation (rs = 0.82) between the methods. While developing the system we identified three amino acids which were mutated in the Vero E6 cell culture adapted PUUV prototype Sotkamo strain sequence, and changing these residues was critical for expression and neutralizing antibody binding of PUUV glycoproteins.
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detection of antibodies against h5 and h7 strains in birds evaluation of influenza pseudovirus particle Neutralization Tests
Infection ecology & epidemiology, 2014Co-Authors: Sofie Wallerstrom, Nina Lagerqvist, Nigel J Temperton, Michaela Cassmer, Malin Karlsson, Mikael Leijon, Ake Lundkvist, Ana Moreno, Kerstin I FalkAbstract:Introduction : Avian influenza viruses circulate in bird populations, and it is important to maintain and uphold our knowledge of the viral strains that are currently of interest in this context. Here, we describe the use of hemagglutinin-pseudotype retroviruses based on highly pathogenic influenza viruses for the screening of avian sera for influenza A antibodies. Our aim was also to determine whether the pseudovirus Neutralization Tests that we assessed were sensitive and simple to use compared to the traditional methods, including hemagglutination inhibition assays and microNeutralization Tests. Material and methods : H5 and H7 pseudovirus Neutralization Tests were evaluated by using serum from infected rabbits. Subsequently, the assays were further investigated using a panel of serum samples from avian species. The panel contained samples that were seropositive for five different hemagglutinin subtypes as well as influenza A seronegative samples. Results and discussion : The results suggest that the pseudovirus Neutralization test is an alternative to hemagglutination inhibition assays, as we observed comparable titers to those of both standard microNeutralizations assays as well as hemagglutinin inhibition assays. When evaluated by a panel of avian sera, the method also showed its capability to recognize antibodies directed toward low-pathogenic H5 and H7. Hence, we conclude that it is possible to use pseudoviruses based on highly pathogenic avian influenza viruses to screen avian sera for antibodies directed against influenza A subtypes H5 and H7. Keywords : influenza A; pseudovirus; Neutralization; antibodies; avian (Published: 15 January 2014) Citation: Infection Ecology and Epidemiology 2014, 4 : 23011 - http://dx.doi.org/10.3402/iee.v4.23011
Tomas Strandin - One of the best experts on this subject based on the ideXlab platform.
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vaccinia virus free rescue of fluorescent replication defective vesicular stomatitis virus and pseudotyping with puumala virus glycoproteins for use in Neutralization Tests
Journal of General Virology, 2016Co-Authors: Rommel Paneth Iheozorejiofor, Ake Lundkvist, Lev Levanov, Jussi Hepojoki, Tomas Strandin, Alexander Plyusnin, Olli VapalahtiAbstract:Puumala virus (PUUV) grows slowly in cell culture. To study antigenic properties of PUUV, an amenable method for their expression would be beneficial. To achieve this, a replication-defective recombinant vesicular stomatitis virus, rVSVΔG*EGFP, was rescued using BSRT7/5 and encephalomyocarditis virus (EMCV) internal ribosomal entry site (IRES)-enabled rescue plasmids. Using these particles, pseudotypes bearing PUUV Sotkamo strain glycoproteins were produced, with titres in the range 105–108, and were used in pseudotype focus reduction Neutralization Tests (pFRNTs) with neutralizing monoclonal antibodies and patient sera. The results were compared with those from orthodox focus reduction Neutralization Tests (oFRNTs) using native PUUV with the same samples and showed a strong positive correlation (rs = 0.82) between the methods. While developing the system we identified three amino acids which were mutated in the Vero E6 cell culture adapted PUUV prototype Sotkamo strain sequence, and changing these residues was critical for expression and neutralizing antibody binding of PUUV glycoproteins.