Dengue Reporter Virus Particles for Measuring Neutralizing Antibodies against Each of the Four Dengue Serotypes
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{"title"=>"Dengue reporter virus particles for measuring neutralizing antibodies against each of the four dengue serotypes", "type"=>"journal", "authors"=>[{"first_name"=>"Kimberly", "last_name"=>"Mattia", "scopus_author_id"=>"54408786400"}, {"first_name"=>"Bridget A.", "last_name"=>"Puffer", "scopus_author_id"=>"6602414258"}, {"first_name"=>"Katherine L.", "last_name"=>"Williams", "scopus_author_id"=>"53865786000"}, {"first_name"=>"Ritela", "last_name"=>"Gonzalez", "scopus_author_id"=>"54408352100"}, {"first_name"=>"Meredith", "last_name"=>"Murray", "scopus_author_id"=>"36714020400"}, {"first_name"=>"Emily", "last_name"=>"Sluzas", "scopus_author_id"=>"16204240400"}, {"first_name"=>"Dan", "last_name"=>"Pagano", "scopus_author_id"=>"56279032800"}, {"first_name"=>"Sandya", "last_name"=>"Ajith", "scopus_author_id"=>"14046483100"}, {"first_name"=>"Megan", "last_name"=>"Bower", "scopus_author_id"=>"12769616900"}, {"first_name"=>"Eli", "last_name"=>"Berdougo", "scopus_author_id"=>"6507495536"}, {"first_name"=>"Eva", "last_name"=>"Harris", "scopus_author_id"=>"7403257174"}, {"first_name"=>"Benjamin J.", "last_name"=>"Doranz", "scopus_author_id"=>"6603437820"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-80655132058", "sgr"=>"80655132058", "pui"=>"362888161", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"22096543", "doi"=>"10.1371/journal.pone.0027252"}, "id"=>"c910559a-0d79-32b6-97d3-8e83f4a681bd", "abstract"=>"The lack of reliable, high-throughput tools for characterizing anti-dengue virus (DENV) antibodies in large numbers of serum samples has been an obstacle in understanding the impact of neutralizing antibodies on disease progression and vaccine efficacy. A reporter system using pseudoinfectious DENV reporter virus particles (RVPs) was previously developed by others to facilitate the genetic manipulation and biological characterization of DENV virions. In the current study, we demonstrate the diagnostic utility of DENV RVPs for measuring neutralizing antibodies in human serum samples against all four DENV serotypes, with attention to the suitability of DENV RVPs for large-scale, long-term studies. DENV RVPs used against human sera yielded serotype-specific responses and reproducible neutralization titers that were in statistical agreement with Plaque Reduction Neutralization Test (PRNT) results. DENV RVPs were also used to measure neutralization titers against the four DENV serotypes in a panel of human sera from a clinical study of dengue patients. The high-throughput capability, stability, rapidity, and reproducibility of assays using DENV RVPs offer advantages for detecting immune responses that can be applied to large-scale clinical studies of DENV infection and vaccination.", "link"=>"http://www.mendeley.com/research/dengue-reporter-virus-particles-measuring-neutralizing-antibodies-against-each-four-dengue-serotypes", "reader_count"=>83, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>27, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>22, "Student > Postgraduate"=>3, "Other"=>8, "Student > Master"=>11, "Student > Bachelor"=>5, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>27, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>22, "Student > Postgraduate"=>3, "Other"=>8, "Student > Master"=>11, "Student > Bachelor"=>5, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>5, "Mathematics"=>1, "Medicine and Dentistry"=>15, "Agricultural and Biological Sciences"=>46, "Arts and Humanities"=>1, "Sports and Recreations"=>1, "Veterinary Science and Veterinary Medicine"=>2, "Chemistry"=>2, "Social Sciences"=>1, "Immunology and Microbiology"=>6}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>15}, "Chemistry"=>{"Chemistry"=>2}, "Social Sciences"=>{"Social Sciences"=>1}, "Sports and Recreations"=>{"Sports and Recreations"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>6}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>46}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Mathematics"=>{"Mathematics"=>1}, "Unspecified"=>{"Unspecified"=>2}, "Arts and Humanities"=>{"Arts and Humanities"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>2}}, "reader_count_by_country"=>{"United States"=>3, "Philippines"=>1, "Brazil"=>1, "Mexico"=>1, "United Kingdom"=>1, "France"=>1, "Germany"=>1, "Indonesia"=>1, "French Polynesia"=>1}, "group_count"=>7}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/715044"], "description"=>"<p>DENV RVPs representing each of the four serotypes were pre-incubated for 1 hour with a WHO reference panel of human primary sera reactive to each RVP serotype (monovalent, squares: anti-DENV-1 serum in <b>A</b>; anti-DENV-2 in <b>B</b>; anti-DENV-3 in <b>C</b>; anti-DENV-4 in <b>D</b>) or reactive to all serotypes (tetravalent, triangles in all four panels). Naïve serum was used as a negative control. After incubation, DENV RVPs were used to infect Raji DC-SIGN-R cells. Forty-eight hours after infection, cells were quantified for GFP expression by flow cytometry (n = 3, error bars represent the standard deviation).</p>", "links"=>[], "tags"=>["denv", "rvps"], "article_id"=>385393, "categories"=>["Virology"], "users"=>["Kimberly Mattia", "Bridget A. Puffer", "Katherine L. Williams", "Ritela Gonzalez", "Meredith Murray", "Emily Sluzas", "Dan Pagano", "Sandya Ajith", "Megan Bower", "Eli Berdougo", "Eva Harris", "Benjamin J. Doranz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027252.g004", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Neutralization_of_DENV_RVPs_using_WHO_human_reference_sera_/385393", "title"=>"Neutralization of DENV RVPs using WHO human reference sera.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-09 01:29:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/715192"], "description"=>"<p>Longitudinal serum samples taken at 3, 6, 12 and 18 months post-symptom onset from two primary DENV-1, one primary DENV-2, two primary DENV-3, and one secondary DENV-2 infection were obtained from naturally infected patients in Nicaragua. The serum representing secondary infection was derived from a patient that was last infected by DENV-2, as determined by RT-PCR and virus isolation. Each serum was used in neutralization assays against each DENV serotype. ‘<10’ (non-neutralizing) indicates failure of the serum to neutralize by at least 50%. Each sample was run once, in duplicate. DF, Dengue Fever; DHF, Dengue Hemorrhagic Fever. ND = not determined.</p>", "links"=>[], "tags"=>["denv", "rvps", "serum"], "article_id"=>385540, "categories"=>["Virology"], "users"=>["Kimberly Mattia", "Bridget A. Puffer", "Katherine L. Williams", "Ritela Gonzalez", "Meredith Murray", "Emily Sluzas", "Dan Pagano", "Sandya Ajith", "Megan Bower", "Eli Berdougo", "Eva Harris", "Benjamin J. Doranz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027252.t002", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Application_of_DENV_RVPs_to_clinical_serum_samples_/385540", "title"=>"Application of DENV RVPs to clinical serum samples.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-11-09 01:32:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/714985"], "description"=>"<p>(<b>A</b>) Serial dilutions of a single lot of DENV-2 RVPs were pre-incubated with the monoclonal antibody 4G2 at room temperature for 1 hour followed by infection of Raji DC-SIGN-R cells. Forty-eight hours after infection, cells were analyzed for GFP expression by flow cytometry (n = 8 for each dilution, error bars represent the standard deviation). (<b>B</b>) NT<sub>50</sub> values for each replicate for the indicated RVP input were calculated and plotted (the bar represents the mean NT<sub>50</sub>, boxes show the mean and standard deviation for each RVP dilution tested). The mean NT<sub>50</sub> values for each volume of RVPs tested for neutralization were not statistically different (ANOVA, p>0.05). (<b>C</b>) A single lot of DENV-2 RVPs was used for independent neutralization experiments performed on four different days in duplicate (mean and range shown). Mean NT<sub>50</sub> values are not statistically different (ANOVA, p>0.05). (<b>D</b>) Independently derived lots of DENV-2 RVPs were used for neutralization experiments performed in duplicate (mean and range shown). Mean NT<sub>50</sub> values are not statistically different across different lots (ANOVA, p>0.05).</p>", "links"=>[], "tags"=>["rvps", "reproducible", "antibody", "neutralization"], "article_id"=>385334, "categories"=>["Virology"], "users"=>["Kimberly Mattia", "Bridget A. Puffer", "Katherine L. Williams", "Ritela Gonzalez", "Meredith Murray", "Emily Sluzas", "Dan Pagano", "Sandya Ajith", "Megan Bower", "Eli Berdougo", "Eva Harris", "Benjamin J. Doranz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027252.g003", "stats"=>{"downloads"=>2, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DENV_RVPs_can_be_used_to_derive_reproducible_antibody_neutralization_titers_/385334", "title"=>"DENV RVPs can be used to derive reproducible antibody neutralization titers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-09 01:28:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/714766"], "description"=>"<p>(<b>A</b>) Dengue Reporter Virus Particles (RVPs) are composed of the proteins capsid (C), premembrane/membrane (prM/M) and envelope (E) from defined DENV strains, and an RNA reporter genome made from a DENV genomic replicon. (<b>B</b>) Serial dilutions of DENV RVPs representing each serotype were used to infect Raji DC-SIGN-R cells. Forty-eight hours after infection, cells were analyzed for GFP expression by flow cytometry. Regression analysis shows a linear relationship (R<sup>2</sup>>0.98 for DENV-1; R<sup>2</sup>>0.99 for DENV-2; R<sup>2</sup>>0.99 for DENV-3; R<sup>2</sup>>0.99 for DENV-4) with the amount (µl) of RVPs used for infection (n = 4, error bars represent the standard deviation). (<b>C</b>) Neutralization assays using DENV RVPs were performed with serially diluted monoclonal antibodies 4G2, 3H5, or the control monoclonal antibody 15F3 for 1 hour prior to infection of Raji DC-SIGN-R cells. Forty-eight hours after infection, cells were quantified for GFP expression by flow cytometry. All neutralization results are shown normalized to the maximum (uninhibited) infection achieved, defined as 100% (n = 3, error bars represent the standard deviation). The infectious titers of the lots of RVPs used here, calculated based on the number of infected cells using an RVP input of 50 µl, are as follows: DENV-1: 773,325 infectious units/ml, DENV-2: 422,970 infectious units/ml, DENV-3: 347,292 infectious units/ml, and DENV-4: 353,995 infectious units/ml.</p>", "links"=>[], "tags"=>["rvps", "infectious", "antigenically", "dengue"], "article_id"=>385112, "categories"=>["Virology"], "users"=>["Kimberly Mattia", "Bridget A. Puffer", "Katherine L. Williams", "Ritela Gonzalez", "Meredith Murray", "Emily Sluzas", "Dan Pagano", "Sandya Ajith", "Megan Bower", "Eli Berdougo", "Eva Harris", "Benjamin J. Doranz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027252.g001", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DENV_RVPs_are_infectious_and_antigenically_equivalent_to_live_dengue_virus_/385112", "title"=>"DENV RVPs are infectious and antigenically equivalent to live dengue virus.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-09 01:25:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/714907"], "description"=>"<p>DENV RVPs representing all 4 serotypes were incubated at 4°C (<b>A</b>), 25°C (<b>B</b>), or at 37°C (<b>C</b>) for up to 10 hours, followed by infection of Raji DC-SIGN-R cells. Forty-eight hours post-infection, cells were analyzed for GFP expression by flow cytometry. At all temperatures, DENV RVPs remain highly infectious after a 2-hour incubation (a typical incubation period used in neutralization assays, vertical dashed line). (<b>D</b>) A single preparation of DENV-2 RVPs was aliquoted and frozen at −80°C from 2 days up to 1 year prior to infection of Raji DC-SIGN-R cells. Forty-eight hours after infection, cells were analyzed for GFP expression by flow cytometry. DENV RVPs showed little or no substantial loss of infectivity after long-term cryopreservation (n = 2, error bars represent the range).</p>", "links"=>[], "tags"=>["rvps", "infectious", "conditions", "neutralization", "assays"], "article_id"=>385257, "categories"=>["Virology"], "users"=>["Kimberly Mattia", "Bridget A. Puffer", "Katherine L. Williams", "Ritela Gonzalez", "Meredith Murray", "Emily Sluzas", "Dan Pagano", "Sandya Ajith", "Megan Bower", "Eli Berdougo", "Eva Harris", "Benjamin J. Doranz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027252.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_DENV_RVPs_are_infectious_under_conditions_commonly_used_in_neutralization_assays_and_following_long_term_cryopreservation_/385257", "title"=>"DENV RVPs are infectious under conditions commonly used in neutralization assays and following long-term cryopreservation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-09 01:27:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/715146"], "description"=>"<p>NT<sub>50</sub> neutralization titers for sera were obtained using DENV RVPs or conventional PRNT with live virus and are expressed as reciprocal serum dilutions at which viral infection was inhibited by 50%. All reference sera were obtained from the National Institute of Biological Standards and Controls (NIBSC). RVPa and RVPb values were obtained from independent experiments performed in different laboratories (IM and UCB) and were compared to PRNT values obtained in an independent study conducted by the WHO for validation testing of PRNT for DENV clinical studies <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0027252#pone.0027252-WHO1\" target=\"_blank\">[23]</a>. All sera were tested at least three times. Mean NT<sub>50</sub> values using PRNT and RVPs were not statistically different (ANOVA, p>0.05). ‘<10’ (non-neutralizing) indicates a calculated NT<sub>50</sub> value of <10 or the failure of the sera to neutralize by at least 50%. NT<sub>50</sub> values <10 were calculated as 10.0 for statistical analysis. ND = not determined.</p>", "links"=>[], "tags"=>["neutralization", "titers", "sera", "prnt"], "article_id"=>385498, "categories"=>["Virology"], "users"=>["Kimberly Mattia", "Bridget A. Puffer", "Katherine L. Williams", "Ritela Gonzalez", "Meredith Murray", "Emily Sluzas", "Dan Pagano", "Sandya Ajith", "Megan Bower", "Eli Berdougo", "Eva Harris", "Benjamin J. Doranz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027252.t001", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_neutralization_titers_for_human_sera_obtained_by_PRNT_or_RVPs_/385498", "title"=>"Comparison of neutralization titers for human sera obtained by PRNT or RVPs.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-11-09 01:31:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/362121", "https://ndownloader.figshare.com/files/362165", "https://ndownloader.figshare.com/files/362201", "https://ndownloader.figshare.com/files/362288", "https://ndownloader.figshare.com/files/362337"], "description"=>"<div><p>The lack of reliable, high-throughput tools for characterizing anti-dengue virus (DENV) antibodies in large numbers of serum samples has been an obstacle in understanding the impact of neutralizing antibodies on disease progression and vaccine efficacy. A reporter system using pseudoinfectious DENV reporter virus particles (RVPs) was previously developed by others to facilitate the genetic manipulation and biological characterization of DENV virions. In the current study, we demonstrate the diagnostic utility of DENV RVPs for measuring neutralizing antibodies in human serum samples against all four DENV serotypes, with attention to the suitability of DENV RVPs for large-scale, long-term studies. DENV RVPs used against human sera yielded serotype-specific responses and reproducible neutralization titers that were in statistical agreement with Plaque Reduction Neutralization Test (PRNT) results. DENV RVPs were also used to measure neutralization titers against the four DENV serotypes in a panel of human sera from a clinical study of dengue patients. The high-throughput capability, stability, rapidity, and reproducibility of assays using DENV RVPs offer advantages for detecting immune responses that can be applied to large-scale clinical studies of DENV infection and vaccination.</p> </div>", "links"=>[], "tags"=>["dengue", "particles", "measuring", "neutralizing", "antibodies", "serotypes"], "article_id"=>131582, "categories"=>["Cancer"], "users"=>["Kimberly Mattia", "Bridget A. Puffer", "Katherine L. Williams", "Ritela Gonzalez", "Meredith Murray", "Emily Sluzas", "Dan Pagano", "Sandya Ajith", "Megan Bower", "Eli Berdougo", "Eva Harris", "Benjamin J. Doranz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027252.s001", "https://dx.doi.org/10.1371/journal.pone.0027252.s002", "https://dx.doi.org/10.1371/journal.pone.0027252.s003", "https://dx.doi.org/10.1371/journal.pone.0027252.s004", "https://dx.doi.org/10.1371/journal.pone.0027252.s005"], "stats"=>{"downloads"=>6, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Dengue_Reporter_Virus_Particles_for_Measuring_Neutralizing_Antibodies_against_Each_of_the_Four_Dengue_Serotypes/131582", "title"=>"Dengue Reporter Virus Particles for Measuring Neutralizing Antibodies against Each of the Four Dengue Serotypes", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-11-09 00:26:22"}

PMC Usage Stats | Further Information

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Relative Metric

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