Conservation and Immunogenicity of Novel Antigens in Diverse Isolates of Enterotoxigenic Escherichia coli
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{"title"=>"Conservation and Immunogenicity of Novel Antigens in Diverse Isolates of Enterotoxigenic Escherichia coli", "type"=>"journal", "authors"=>[{"first_name"=>"Qingwei", "last_name"=>"Luo", "scopus_author_id"=>"55544633400"}, {"first_name"=>"Firdausi", "last_name"=>"Qadri", "scopus_author_id"=>"7005341667"}, {"first_name"=>"Rita", "last_name"=>"Kansal", "scopus_author_id"=>"6603851411"}, {"first_name"=>"David A.", "last_name"=>"Rasko", "scopus_author_id"=>"6602145541"}, {"first_name"=>"Alaullah", "last_name"=>"Sheikh", "scopus_author_id"=>"16426258000"}, {"first_name"=>"James M.", "last_name"=>"Fleckenstein", "scopus_author_id"=>"7006968615"}], "year"=>2015, "source"=>"PLoS Neglected Tropical Diseases", "identifiers"=>{"pui"=>"601975975", "issn"=>"19352735", "isbn"=>"19352735", "doi"=>"10.1371/journal.pntd.0003446", "scopus"=>"2-s2.0-84922324379", "pmid"=>"25629897", "sgr"=>"84922324379"}, "id"=>"fd0c572f-180e-3d16-94fb-6034975488a7", "abstract"=>"BACKGROUND: Enterotoxigenic Escherichia coli (ETEC) are common causes of diarrheal morbidity and mortality in developing countries for which there is currently no vaccine. Heterogeneity in classical ETEC antigens known as colonization factors (CFs) and poor efficacy of toxoid-based approaches to date have impeded development of a broadly protective ETEC vaccine, prompting searches for novel molecular targets.\\n\\nMETHODOLOGY: Using a variety of molecular methods, we examined a large collection of ETEC isolates for production of two secreted plasmid-encoded pathotype-specific antigens, the EtpA extracellular adhesin, and EatA, a mucin-degrading serine protease; and two chromosomally-encoded molecules, the YghJ metalloprotease and the EaeH adhesin, that are not specific to the ETEC pathovar, but which have been implicated in ETEC pathogenesis. ELISA assays were also performed on control and convalescent sera to characterize the immune response to these antigens. Finally, mice were immunized with recombinant EtpA (rEtpA), and a protease deficient version of the secreted EatA passenger domain (rEatApH134R) to examine the feasibility of combining these molecules in a subunit vaccine approach.\\n\\nPRINCIPAL FINDINGS: EtpA and EatA were secreted by more than half of all ETEC, distributed over diverse phylogenetic lineages belonging to multiple CF groups, and exhibited surprisingly little sequence variation. Both chromosomally-encoded molecules were also identified in a wide variety of ETEC strains and YghJ was secreted by 89% of isolates. Antibodies against both the ETEC pathovar-specific and conserved E. coli antigens were present in significantly higher titers in convalescent samples from subjects with ETEC infection than controls suggesting that each of these antigens is produced and recognized during infection. Finally, co-immunization of mice with rEtpA and rEatApH134R offered significant protection against ETEC infection.\\n\\nCONCLUSIONS: Collectively, these data suggest that novel antigens could significantly complement current approaches and foster improved strategies for development of broadly protective ETEC vaccines.", "link"=>"http://www.mendeley.com/research/conservation-immunogenicity-novel-antigens-diverse-isolates-enterotoxigenic-escherichia-coli", "reader_count"=>22, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>11, "Student > Ph. D. Student"=>5, "Student > Master"=>2, "Student > Bachelor"=>1, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>11, "Student > Ph. D. Student"=>5, "Student > Master"=>2, "Student > Bachelor"=>1, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>1, "Materials Science"=>1, "Agricultural and Biological Sciences"=>11, "Medicine and Dentistry"=>2, "Veterinary Science and Veterinary Medicine"=>1, "Immunology and Microbiology"=>3}, "reader_count_by_subdiscipline"=>{"Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>11}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>3}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Chile"=>1, "Kenya"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1875086"], "description"=>"<p>Serologic responses to <b>a</b> heat-labile toxin (LT), <b>b</b> the passenger domain of EatA (Eatp), <b>c</b> EtpA. Shown are serum IgG responses following intranasal vaccination of mice with the LT adjuvant alone, or LT with 15 μg of either the proteolytically inactive passenger domain (EatAp<sub>H134R</sub>), EtpA, or both antigens on days 0, 14, 28. Colonization of mice following immunization with the adjuvant alone compared with single and dual antigen vaccination. Comparisons between groups were by Mann Whitney two tailed nonparametric testing. (One mouse died during the vaccination period in the LT/EtpA group and was therefore excluded from the analysis). For mice with no detectable colonies following challenge, the number of cfu is arbitrarily reported as 1 (10°) cfu, the theoretical limit of detection.</p>", "links"=>[], "tags"=>["Enterotoxigenic Escherichia coli BackgroundEnterotoxigenic Escherichia coli", "etec", "rEatApH 134R", "antigen", "subunit vaccine approach.Principal FindingsEtpA", "EtpA extracellular adhesin", "EatA passenger domain", "elisa", "cf"], "article_id"=>1296832, "categories"=>["Biological Sciences"], "users"=>["Qingwei Luo", "Firdausi Qadri", "Rita Kansal", "David A. Rasko", "Alaullah Sheikh", "James M. Fleckenstein"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0003446.g004", "stats"=>{"downloads"=>0, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mice_immunized_with_rEtpA_and_rEatAp_H134R_are_protected_against_ETEC_infection_/1296832", "title"=>"Mice immunized with rEtpA and rEatAp<sub>H134R</sub> are protected against ETEC infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-01-28 03:33:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1875080"], "description"=>"<p>Shown are kinetic ELISA data for four different recombinant antigens (a, rEtpA; b, the rEatA passenger domain; c, rYghJ; and d, rEaeH) obtained with 1:4096 dilutions of convalescent plasma from ETEC-infected patients hospitalized at ICDDR,B in Dhaka, Bangladesh (closed circles), or control patients not infected with enterotoxigenic <i>E. coli</i> (open circles). Horizontal bars represent geometric mean Vmax kinetic ELISA values for each group. P values obtained by two-tailed Mann Whitney testing of groups are summarized (*<0.05; **<0.01; ***<0.001;****<0.0001). x-axis of each graph depicts the specificity of the secondary antibody used in the ELISA (IgG, IgA, IgM, and total IgG, IgA, and IgM).</p>", "links"=>[], "tags"=>["Enterotoxigenic Escherichia coli BackgroundEnterotoxigenic Escherichia coli", "etec", "rEatApH 134R", "antigen", "subunit vaccine approach.Principal FindingsEtpA", "EtpA extracellular adhesin", "EatA passenger domain", "elisa", "cf"], "article_id"=>1296830, "categories"=>["Biological Sciences"], "users"=>["Qingwei Luo", "Firdausi Qadri", "Rita Kansal", "David A. Rasko", "Alaullah Sheikh", "James M. Fleckenstein"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0003446.g003", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Recognition_of_novel_antigens_during_naturally_occurring_ETEC_infections_in_Bangladesh_/1296830", "title"=>"Recognition of novel antigens during naturally occurring ETEC infections in Bangladesh.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-01-28 03:33:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1875094", "https://ndownloader.figshare.com/files/1875095", "https://ndownloader.figshare.com/files/1875096", "https://ndownloader.figshare.com/files/1875097", "https://ndownloader.figshare.com/files/1875098", "https://ndownloader.figshare.com/files/1875099", "https://ndownloader.figshare.com/files/1875100"], "description"=>"<div><p>Background</p><p>Enterotoxigenic <i>Escherichia coli</i> (ETEC) are common causes of diarrheal morbidity and mortality in developing countries for which there is currently no vaccine. Heterogeneity in classical ETEC antigens known as colonization factors (CFs) and poor efficacy of toxoid-based approaches to date have impeded development of a broadly protective ETEC vaccine, prompting searches for novel molecular targets.</p><p>Methodology</p><p>Using a variety of molecular methods, we examined a large collection of ETEC isolates for production of two secreted plasmid-encoded pathotype-specific antigens, the EtpA extracellular adhesin, and EatA, a mucin-degrading serine protease; and two chromosomally-encoded molecules, the YghJ metalloprotease and the EaeH adhesin, that are not specific to the ETEC pathovar, but which have been implicated in ETEC pathogenesis. ELISA assays were also performed on control and convalescent sera to characterize the immune response to these antigens. Finally, mice were immunized with recombinant EtpA (rEtpA), and a protease deficient version of the secreted EatA passenger domain (rEatAp<sub>H134R</sub>) to examine the feasibility of combining these molecules in a subunit vaccine approach.</p><p>Principal Findings</p><p>EtpA and EatA were secreted by more than half of all ETEC, distributed over diverse phylogenetic lineages belonging to multiple CF groups, and exhibited surprisingly little sequence variation. Both chromosomally-encoded molecules were also identified in a wide variety of ETEC strains and YghJ was secreted by 89% of isolates. Antibodies against both the ETEC pathovar-specific and conserved <i>E. coli</i> antigens were present in significantly higher titers in convalescent samples from subjects with ETEC infection than controls suggesting that each of these antigens is produced and recognized during infection. Finally, co-immunization of mice with rEtpA and rEatAp<sub>H134R</sub> offered significant protection against ETEC infection.</p><p>Conclusions</p><p>Collectively, these data suggest that novel antigens could significantly complement current approaches and foster improved strategies for development of broadly protective ETEC vaccines.</p></div>", "links"=>[], "tags"=>["Enterotoxigenic Escherichia coli BackgroundEnterotoxigenic Escherichia coli", "etec", "rEatApH 134R", "antigen", "subunit vaccine approach.Principal FindingsEtpA", "EtpA extracellular adhesin", "EatA passenger domain", "elisa", "cf"], "article_id"=>1296840, "categories"=>["Biological Sciences"], "users"=>["Qingwei Luo", "Firdausi Qadri", "Rita Kansal", "David A. Rasko", "Alaullah Sheikh", "James M. Fleckenstein"], "doi"=>["https://dx.doi.org/10.1371/journal.pntd.0003446.s001", "https://dx.doi.org/10.1371/journal.pntd.0003446.s002", "https://dx.doi.org/10.1371/journal.pntd.0003446.s003", "https://dx.doi.org/10.1371/journal.pntd.0003446.s004", "https://dx.doi.org/10.1371/journal.pntd.0003446.s005", "https://dx.doi.org/10.1371/journal.pntd.0003446.s006", "https://dx.doi.org/10.1371/journal.pntd.0003446.s007"], "stats"=>{"downloads"=>14, "page_views"=>42, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Conservation_and_Immunogenicity_of_Novel_Antigens_in_Diverse_Isolates_of_Enterotoxigenic_Escherichia_coli_/1296840", "title"=>"Conservation and Immunogenicity of Novel Antigens in Diverse Isolates of Enterotoxigenic <i>Escherichia coli</i>", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2015-01-28 03:33:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1875089"], "description"=>"<p>EtpA sequence conservation in geographically, temporally, and phylogenically disparate isolates.</p>", "links"=>[], "tags"=>["Enterotoxigenic Escherichia coli BackgroundEnterotoxigenic Escherichia coli", "etec", "rEatApH 134R", "antigen", "subunit vaccine approach.Principal FindingsEtpA", "EtpA extracellular adhesin", "EatA passenger domain", "elisa", "cf"], "article_id"=>1296835, "categories"=>["Biological Sciences"], "users"=>["Qingwei Luo", "Firdausi Qadri", "Rita Kansal", "David A. Rasko", "Alaullah Sheikh", "James M. Fleckenstein"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0003446.t003", "stats"=>{"downloads"=>0, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_EtpA_sequence_conservation_in_geographically_temporally_and_phylogenically_disparate_isolates_/1296835", "title"=>"EtpA sequence conservation in geographically, temporally, and phylogenically disparate isolates.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-01-28 03:33:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1875088"], "description"=>"<p>EatA sequence conservation in geographically, temporally, and phylogenically disparate isolates.</p>", "links"=>[], "tags"=>["Enterotoxigenic Escherichia coli BackgroundEnterotoxigenic Escherichia coli", "etec", "rEatApH 134R", "antigen", "subunit vaccine approach.Principal FindingsEtpA", "EtpA extracellular adhesin", "EatA passenger domain", "elisa", "cf"], "article_id"=>1296834, "categories"=>["Biological Sciences"], "users"=>["Qingwei Luo", "Firdausi Qadri", "Rita Kansal", "David A. Rasko", "Alaullah Sheikh", "James M. Fleckenstein"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0003446.t002", "stats"=>{"downloads"=>7, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_EatA_sequence_conservation_in_geographically_temporally_and_phylogenically_disparate_isolates_/1296834", "title"=>"EatA sequence conservation in geographically, temporally, and phylogenically disparate isolates.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-01-28 03:33:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1875079"], "description"=>"<p>a. Heatmap of EtpA and EatA showing the proportion of strains positive for expression of these antigens among different CF groups. CF antigen designation is shown at left of the heatmap. nd = no CF antigen detected. Below is the heatmap key depicting colors associated with each degree of antigen positivity. Density line in yellow depicts the relative number of map features assigned at each proportion. Primary data used to construct the heatmap can be found in <a href=\"http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003446#pntd.0003446.s007\" target=\"_blank\">S2 Dataset</a>. b. Imunoblot detection of EtpA and EatA expression among strains from different phylogenies. The upper immunoblot demonstrates EtpA production in the prototype H10407 strain, ThroopD isolated in Dallas, TX in 1975, the Juruá_18/11 (Amazon, 1998), and phylogentically dispersed strains from icddr,b. The <i>etpA</i> mutant is included as a negative control. The lower blot demonstrates EatA production by H10407, phylogenically distributed strains from icddr,b and Envira_10/1, an additional isolate from cholera-like outbreaks in the Amazon. The <i>eatA</i> mutant is included as a negative control. c. Phlyogram showing the phylogenetic distribution of selected ETEC strains (designations in blue) and reference E. coli strains (designations in black). Red circles and gold stars represent eatA+, and etpA+ strains, respectively.</p>", "links"=>[], "tags"=>["Enterotoxigenic Escherichia coli BackgroundEnterotoxigenic Escherichia coli", "etec", "rEatApH 134R", "antigen", "subunit vaccine approach.Principal FindingsEtpA", "EtpA extracellular adhesin", "EatA passenger domain", "elisa", "cf"], "article_id"=>1296829, "categories"=>["Biological Sciences"], "users"=>["Qingwei Luo", "Firdausi Qadri", "Rita Kansal", "David A. Rasko", "Alaullah Sheikh", "James M. Fleckenstein"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0003446.g002", "stats"=>{"downloads"=>3, "page_views"=>30, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Conservation_of_novel_pathotype_specific_antigens_EtpA_and_EatA_among_phylogenically_distinct_strains_expressing_different_colonization_factors_/1296829", "title"=>"Conservation of novel pathotype-specific antigens EtpA and EatA among phylogenically distinct strains expressing different colonization factors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-01-28 03:33:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1875074"], "description"=>"<p>All of the strains in the collection (n = 181) were examined for production of three secreted ETEC virulence proteins EtpA, EatA, and YghJ by immunoblotting of culture supernatants with the respective antibodies. A subset of these strains (n = 91) were recently sequenced at the Genome Sequencing Center for Infectious Diseases (GSCID).</p>", "links"=>[], "tags"=>["Enterotoxigenic Escherichia coli BackgroundEnterotoxigenic Escherichia coli", "etec", "rEatApH 134R", "antigen", "subunit vaccine approach.Principal FindingsEtpA", "EtpA extracellular adhesin", "EatA passenger domain", "elisa", "cf"], "article_id"=>1296824, "categories"=>["Biological Sciences"], "users"=>["Qingwei Luo", "Firdausi Qadri", "Rita Kansal", "David A. Rasko", "Alaullah Sheikh", "James M. Fleckenstein"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0003446.g001", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Relationship_of_strain_subsets_used_in_antigen_expression_studies_and_strains_with_available_whole_genome_sequences_/1296824", "title"=>"Relationship of strain subsets used in antigen expression studies, and strains with available whole genome sequences.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-01-28 03:33:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/1875087"], "description"=>"<p>Distribution of EtpA and EatA among strains expressing different colonization factors.</p>", "links"=>[], "tags"=>["Enterotoxigenic Escherichia coli BackgroundEnterotoxigenic Escherichia coli", "etec", "rEatApH 134R", "antigen", "subunit vaccine approach.Principal FindingsEtpA", "EtpA extracellular adhesin", "EatA passenger domain", "elisa", "cf"], "article_id"=>1296833, "categories"=>["Biological Sciences"], "users"=>["Qingwei Luo", "Firdausi Qadri", "Rita Kansal", "David A. Rasko", "Alaullah Sheikh", "James M. Fleckenstein"], "doi"=>"https://dx.doi.org/10.1371/journal.pntd.0003446.t001", "stats"=>{"downloads"=>5, "page_views"=>21, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Distribution_of_EtpA_and_EatA_among_strains_expressing_different_colonization_factors_/1296833", "title"=>"Distribution of EtpA and EatA among strains expressing different colonization factors.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2015-01-28 03:33:47"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2015-01-01T00:00:00Z", "end_date"=>"2015-12-31T00:00:00Z", "subject_areas"=>[]}
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