A Neutralizing Monoclonal Antibody Targeting the Acid-Sensitive Region in Chikungunya Virus E2 Protects from Disease
Publication Date
September 12, 2013
Journal
PLOS Neglected Tropical Diseases
Authors
Suganya Selvarajah, Nicole R. Sexton, Kristen M. Kahle, Rachel H. Fong, et al
Volume
7
Issue
9
Pages
e2423
DOI
https://dx.plos.org/10.1371/journal.pntd.0002423
Publisher URL
http://journals.plos.org/plosntds/article?id=10.1371%2Fjournal.pntd.0002423
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/24069479
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3772074
Europe PMC
http://europepmc.org/abstract/MED/24069479
Web of Science
000324920800028
Scopus
84884683466
Mendeley
http://www.mendeley.com/research/neutralizing-monoclonal-antibody-targeting-acidsensitive-region-chikungunya-virus-e2-protects-diseas
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Mendeley | Further Information

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CrossRef

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1203127", "https://ndownloader.figshare.com/files/1203129", "https://ndownloader.figshare.com/files/1203130"], "description"=>"<div><p>The mosquito-borne alphavirus, chikungunya virus (CHIKV), has recently reemerged, producing the largest epidemic ever recorded for this virus, with up to 6.5 million cases of acute and chronic rheumatic disease. There are currently no licensed vaccines for CHIKV and current anti-inflammatory drug treatment is often inadequate. Here we describe the isolation and characterization of two human monoclonal antibodies, C9 and E8, from CHIKV infected and recovered individuals. C9 was determined to be a potent virus neutralizing antibody and a biosensor antibody binding study demonstrated it recognized residues on intact CHIKV VLPs. Shotgun mutagenesis alanine scanning of 98 percent of the residues in the E1 and E2 glycoproteins of CHIKV envelope showed that the epitope bound by C9 included amino-acid 162 in the acid-sensitive region (ASR) of the CHIKV E2 glycoprotein. The ASR is critical for the rearrangement of CHIKV E2 during fusion and viral entry into host cells, and we predict that C9 prevents these events from occurring. When used prophylactically in a CHIKV mouse model, C9 completely protected against CHIKV viremia and arthritis. We also observed that when administered therapeutically at 8 or 18 hours post-CHIKV challenge, C9 gave 100% protection in a pathogenic mouse model. Given that targeting this novel neutralizing epitope in E2 can potently protect both in vitro and in vivo, it is likely to be an important region both for future antibody and vaccine-based interventions against CHIKV.</p></div>", "links"=>[], "tags"=>["microbiology", "Virology", "Viral structure", "Viral envelope", "Animal models of infection", "antivirals", "Emerging viral diseases", "Viral vaccines", "Infectious diseases", "neglected tropical diseases", "Arboviral infections", "neutralizing", "monoclonal", "antibody", "targeting", "acid-sensitive", "chikungunya", "e2", "protects"], "article_id"=>798445, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Suganya Selvarajah", "Nicole R. Sexton", "Kristen M. Kahle", "Rachel H. Fong", "Kimberly-Anne Mattia", "Joy Gardner", "Kai Lu", "Nathan M. Liss", "Beatriz Salvador", "David F. Tucker", "Trevor Barnes", "Manu Mabila", "Xiangdong Zhou", "Giada Rossini", "Joseph B. Rucker", "David Avram Sanders", "Andreas Suhrbier", "Vittorio Sambri", "Alain Michault", "Marcus O. Muench", "Benjamin J. Doranz", "Graham Simmons"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0002423.s001", "https://dx.doi.org/10.1371/journal.pntd.0002423.s002", "https://dx.doi.org/10.1371/journal.pntd.0002423.s003"], "stats"=>{"downloads"=>5, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_A_Neutralizing_Monoclonal_Antibody_Targeting_the_Acid_Sensitive_Region_in_Chikungunya_Virus_E2_Protects_from_Disease_/798445", "title"=>"A Neutralizing Monoclonal Antibody Targeting the Acid-Sensitive Region in Chikungunya Virus E2 Protects from Disease", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-09-12 03:06:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1203118"], "description"=>"1<p>All mice were infected with CHIKV at hour 0. Data on CHIKV infected mice without antibody treatment and mice treated with control or C9 antibody at hour 0 are also shown in <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002423#pntd-0002423-g006\" target=\"_blank\">Figure 6</a>.</p>2<p><i>P</i>-values represent all possible comparisons between C9 treated mice and untreated controls or human IgG treated animals treated with human IgG at hours 0 or +18. C9 treatment significantly affected survival in all cases.</p>", "links"=>[], "tags"=>["microbiology", "Virology", "Viral structure", "Viral envelope", "Animal models of infection", "antivirals", "Emerging viral diseases", "Viral vaccines", "Infectious diseases", "neglected tropical diseases", "Arboviral infections", "chikv-driven", "lethality", "neonate"], "article_id"=>798438, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Suganya Selvarajah", "Nicole R. Sexton", "Kristen M. Kahle", "Rachel H. Fong", "Kimberly-Anne Mattia", "Joy Gardner", "Kai Lu", "Nathan M. Liss", "Beatriz Salvador", "David F. Tucker", "Trevor Barnes", "Manu Mabila", "Xiangdong Zhou", "Giada Rossini", "Joseph B. Rucker", "David Avram Sanders", "Andreas Suhrbier", "Vittorio Sambri", "Alain Michault", "Marcus O. Muench", "Benjamin J. Doranz", "Graham Simmons"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0002423.t001", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Therapeutic_protection_against_CHIKV_driven_lethality_in_neonate_mice_1_/798438", "title"=>"Therapeutic protection against CHIKV-driven lethality in neonate mice<sup>1</sup>.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-09-12 03:06:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1203115"], "description"=>"<p>(<b>A)</b> HEK-293 cells expressing mutant CHIKV envelope proteins were immunostained with C9 antibody. Clones with reactivity <20% relative to wild-type CHIKV env were identified as critical for C9 binding. Mutation of residue A162 in E2 to serine significantly reduced C9 binding (green bar) but did not affect binding of E8 (red bar) or other control antibodies (gray bars). Residues are numbered according to E2 in PDB entry #3N41 <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002423#pntd.0002423-Voss1\" target=\"_blank\">[15]</a>. (<b>B</b>) A162S and A162V pseudoviruses were tested for C9 inhibitory potency. The infectivity of the mutants compared to WT was tested (inset graph), indicating that the mutants did not hinder CHIKV env folding or function. Average raw luminescence units are shown for each construct and an env-minus negative control. (<b>C</b>) The critical residue A162 (shown in green) was visualized on the CHIKV env crystal structure. The E1, E2, and E3 env protein subunits in the monomer (PDB Entry #3N41) are depicted in yellow, red, and blue, respectively and the fusion loop is shown in silver (left panel). In the side-view and top-down trimeric representations (center and right panels, PDB entry #2XFC), E3 is not in the structure. In the side view trimeric representation (center panel), the viral membrane is positioned at the bottom of the figure.</p>", "links"=>[], "tags"=>["microbiology", "Virology", "Viral structure", "Viral envelope", "Animal models of infection", "antivirals", "Emerging viral diseases", "Viral vaccines", "Infectious diseases", "neglected tropical diseases", "Arboviral infections", "residues", "c9", "binding"], "article_id"=>798435, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Suganya Selvarajah", "Nicole R. Sexton", "Kristen M. Kahle", "Rachel H. Fong", "Kimberly-Anne Mattia", "Joy Gardner", "Kai Lu", "Nathan M. Liss", "Beatriz Salvador", "David F. Tucker", "Trevor Barnes", "Manu Mabila", "Xiangdong Zhou", "Giada Rossini", "Joseph B. Rucker", "David Avram Sanders", "Andreas Suhrbier", "Vittorio Sambri", "Alain Michault", "Marcus O. Muench", "Benjamin J. Doranz", "Graham Simmons"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0002423.g004", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Critical_residues_and_predicted_C9_binding_site_/798435", "title"=>"Critical residues and predicted C9 binding site.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-12 03:06:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1203109"], "description"=>"<p>Neutralization of pseudovirus bearing CHIKV S27 wild-type (black); CHIKV A226V mutant (light blue); SFV (red); SINV (magenta); RRV (green) and VSV (dark blue) envelope by (<b>A</b>) C9 or (<b>B</b>) E8 mAbs. Antibody concentration is shown in the x-axis. The results are expressed as the percentage of no antibody control and represent mean of triplicate wells, and is representative of three experiments.</p>", "links"=>[], "tags"=>["microbiology", "Virology", "Viral structure", "Viral envelope", "Animal models of infection", "antivirals", "Emerging viral diseases", "Viral vaccines", "Infectious diseases", "neglected tropical diseases", "Arboviral infections", "mabs", "c9", "e8", "neutralize", "chikv"], "article_id"=>798429, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Suganya Selvarajah", "Nicole R. Sexton", "Kristen M. Kahle", "Rachel H. Fong", "Kimberly-Anne Mattia", "Joy Gardner", "Kai Lu", "Nathan M. Liss", "Beatriz Salvador", "David F. Tucker", "Trevor Barnes", "Manu Mabila", "Xiangdong Zhou", "Giada Rossini", "Joseph B. Rucker", "David Avram Sanders", "Andreas Suhrbier", "Vittorio Sambri", "Alain Michault", "Marcus O. Muench", "Benjamin J. Doranz", "Graham Simmons"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0002423.g001", "stats"=>{"downloads"=>0, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Human_mAbs_C9_and_E8_neutralize_CHIKV_pseudovirions_/798429", "title"=>"Human mAbs C9 and E8 neutralize CHIKV pseudovirions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-12 03:06:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1203117"], "description"=>"<p>C57BL/6J neonatal mice were injected with 5×10<sup>5</sup> PFU CHIKV (isolate S27) virus. Co-incident with infection, groups of mice (n = 5 to 8 mice per group) were inoculated with C9 mAb at the indicated concentrations or human IgG as control. The protective nature of C9 mAb at different concentrations is represented as Kaplan-Meier survival curves.</p>", "links"=>[], "tags"=>["microbiology", "Virology", "Viral structure", "Viral envelope", "Animal models of infection", "antivirals", "Emerging viral diseases", "Viral vaccines", "Infectious diseases", "neglected tropical diseases", "Arboviral infections", "c9", "mab", "neonate"], "article_id"=>798437, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Suganya Selvarajah", "Nicole R. Sexton", "Kristen M. Kahle", "Rachel H. Fong", "Kimberly-Anne Mattia", "Joy Gardner", "Kai Lu", "Nathan M. Liss", "Beatriz Salvador", "David F. Tucker", "Trevor Barnes", "Manu Mabila", "Xiangdong Zhou", "Giada Rossini", "Joseph B. Rucker", "David Avram Sanders", "Andreas Suhrbier", "Vittorio Sambri", "Alain Michault", "Marcus O. Muench", "Benjamin J. Doranz", "Graham Simmons"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0002423.g006", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Therapeutic_protection_by_C9_mAb_of_neonate_mice_/798437", "title"=>"Therapeutic protection by C9 mAb of neonate mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-12 03:06:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1203116"], "description"=>"<p>C57BL/6 mice (n = 4 mice per group) were injected with (i) PBS; (ii) purified C9 mAb; or (iii) purified control human mAb at 0.5 mg/mouse by the intraperitoneal route one day (day −1) prior to infection on day 0 with CHIKV (isolate LR2006-OPY1). (<b>A</b>) Peripheral blood viremia (CCID<sub>50</sub>/ml). X-axis represents days post-CHIKV inoculation; (<b>B</b>) Foot swelling over time is presented as a group average of the percentage increase in foot height×width (in the metatarsal region) for each foot compared with the same foot on day 0 (n = 8 feet).</p>", "links"=>[], "tags"=>["microbiology", "Virology", "Viral structure", "Viral envelope", "Animal models of infection", "antivirals", "Emerging viral diseases", "Viral vaccines", "Infectious diseases", "neglected tropical diseases", "Arboviral infections", "arthritis", "viremia", "chikv"], "article_id"=>798436, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Suganya Selvarajah", "Nicole R. Sexton", "Kristen M. Kahle", "Rachel H. Fong", "Kimberly-Anne Mattia", "Joy Gardner", "Kai Lu", "Nathan M. Liss", "Beatriz Salvador", "David F. Tucker", "Trevor Barnes", "Manu Mabila", "Xiangdong Zhou", "Giada Rossini", "Joseph B. Rucker", "David Avram Sanders", "Andreas Suhrbier", "Vittorio Sambri", "Alain Michault", "Marcus O. Muench", "Benjamin J. Doranz", "Graham Simmons"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0002423.g005", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Protection_against_arthritis_and_viremia_in_a_CHIKV_mouse_model_/798436", "title"=>"Protection against arthritis and viremia in a CHIKV mouse model.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-12 03:06:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1203113"], "description"=>"<p>(<b>A</b>) HEK-293 cells expressing mutant CHIKV envelope proteins were immunostained with E8 antibody. Clones with reactivity <20% relative to wild-type CHIKV env were identified as critical for E8 binding. Mutation of six individual E2 residues to alanine (Y69, F84, V113, G114, T116, and D117) significantly reduced E8 binding (red bars) but did not affect binding of C9 (green bar) or other control antibodies (gray bars). Residues are numbered according to E2 in PDB entry #3N41 <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002423#pntd.0002423-Voss1\" target=\"_blank\">[15]</a>. (<b>B</b>) Critical binding residues for E8 (shown in green) were visualized on the CHIKV env crystal structure. The E1, E2, and E3 envelope protein subunits in the monomer (PDB Entry #3N41) are depicted in yellow, red, and blue, respectively and the fusion loop is shown in silver (left panel). In the side-view and top-down trimeric representations (center, and right panels, PDB entry #2XFC), E3 is not in the structure. In the side view trimeric representation (center panel), the viral membrane is positioned at the bottom of the figure.</p>", "links"=>[], "tags"=>["microbiology", "Virology", "Viral structure", "Viral envelope", "Animal models of infection", "antivirals", "Emerging viral diseases", "Viral vaccines", "Infectious diseases", "neglected tropical diseases", "Arboviral infections", "residues", "e8", "binding"], "article_id"=>798433, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Suganya Selvarajah", "Nicole R. Sexton", "Kristen M. Kahle", "Rachel H. Fong", "Kimberly-Anne Mattia", "Joy Gardner", "Kai Lu", "Nathan M. Liss", "Beatriz Salvador", "David F. Tucker", "Trevor Barnes", "Manu Mabila", "Xiangdong Zhou", "Giada Rossini", "Joseph B. Rucker", "David Avram Sanders", "Andreas Suhrbier", "Vittorio Sambri", "Alain Michault", "Marcus O. Muench", "Benjamin J. Doranz", "Graham Simmons"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0002423.g003", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Critical_residues_and_predicted_E8_binding_site_/798433", "title"=>"Critical residues and predicted E8 binding site.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-12 03:06:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/1203111"], "description"=>"<p>(<b>A</b>) Summary of antibody/antigen interactions. Binding of anti-CHIKV antibody C9 to intact CHIKV VLPs was detected using the FoteBio OctetRed biosensor. C9 mAb (66.6 nM) binding to CHIKV VLPs or a non-particle surface control is used to show binding specificity of mAb to intact CHIKV VLP. (<b>B</b>) C9 dose response curve for binding intact CHIKV VLPs. Raw data curves for antibody associating and dissociating from captured CHIKV VLPs are shown in black and fitted curves are shown in red. Data were fitted to a 1∶1 binding model to determine association rate (k<sub>on</sub>) and dissociation rate (k<sub>off</sub>), and equilibrium binding affinity (K<sub>D</sub>) was calculated. C9 binds CHIKV VLPs with 1.21 nM apparent affinity.</p>", "links"=>[], "tags"=>["microbiology", "Virology", "Viral structure", "Viral envelope", "Animal models of infection", "antivirals", "Emerging viral diseases", "Viral vaccines", "Infectious diseases", "neglected tropical diseases", "Arboviral infections", "binding", "chikv"], "article_id"=>798431, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Suganya Selvarajah", "Nicole R. Sexton", "Kristen M. Kahle", "Rachel H. Fong", "Kimberly-Anne Mattia", "Joy Gardner", "Kai Lu", "Nathan M. Liss", "Beatriz Salvador", "David F. Tucker", "Trevor Barnes", "Manu Mabila", "Xiangdong Zhou", "Giada Rossini", "Joseph B. Rucker", "David Avram Sanders", "Andreas Suhrbier", "Vittorio Sambri", "Alain Michault", "Marcus O. Muench", "Benjamin J. Doranz", "Graham Simmons"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0002423.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Kinetic_analysis_of_binding_to_intact_CHIKV_virus_like_particles_/798431", "title"=>"Kinetic analysis of binding to intact CHIKV virus like particles.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-09-12 03:06:12"}

PMC Usage Stats | Further Information

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Relative Metric

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