Targeting Cattle-Borne Zoonoses and Cattle Pathogens Using a Novel Trypanosomatid-Based Delivery System
Publication Date
October 27, 2011
Journal
PLOS Pathogens
Authors
G. Adam Mott, Raymond Wilson, Anuruddika Fernando, Ailie Robinson, et al
Volume
7
Issue
10
Pages
e1002340
DOI
https://dx.plos.org/10.1371/journal.ppat.1002340
Publisher URL
http://journals.plos.org/plospathogens/article?id=10.1371%2Fjournal.ppat.1002340
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/22046137
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3203185
Europe PMC
http://europepmc.org/abstract/MED/22046137
Web of Science
000296734300060
Scopus
80055098912
Mendeley
http://www.mendeley.com/research/targeting-cattleborne-zoonoses-cattle-pathogens-using-novel-trypanosomatidbased-delivery-system
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Mendeley 6526 Apr 04:10 UTC

Mendeley | Further Information

{"title"=>"Targeting cattle-borne zoonoses and cattle pathogens using a novel trypanosomatid-based delivery system", "type"=>"journal", "authors"=>[{"first_name"=>"G. Adam", "last_name"=>"Mott", "scopus_author_id"=>"25936542500"}, {"first_name"=>"Raymond", "last_name"=>"Wilson", "scopus_author_id"=>"57199296369"}, {"first_name"=>"Anuruddika", "last_name"=>"Fernando", "scopus_author_id"=>"54402606200"}, {"first_name"=>"Ailie", "last_name"=>"Robinson", "scopus_author_id"=>"44561475700"}, {"first_name"=>"Paula", "last_name"=>"MacGregor", "scopus_author_id"=>"23100347500"}, {"first_name"=>"David", "last_name"=>"Kennedy", "scopus_author_id"=>"7403112675"}, {"first_name"=>"Dick", "last_name"=>"Schaap", "scopus_author_id"=>"7003611527"}, {"first_name"=>"Jacqueline B.", "last_name"=>"Matthews", "scopus_author_id"=>"25422557800"}, {"first_name"=>"Keith R.", "last_name"=>"Matthews", "scopus_author_id"=>"7201620709"}], "year"=>2011, "source"=>"PLoS Pathogens", "identifiers"=>{"sgr"=>"80055098912", "pmid"=>"22046137", "isbn"=>"1553-7374 (Electronic)\\r1553-7366 (Linking)", "scopus"=>"2-s2.0-80055098912", "issn"=>"15537366", "pui"=>"362834541", "doi"=>"10.1371/journal.ppat.1002340"}, "id"=>"82c1b8b2-14d2-3558-923a-fc06b83f3df6", "abstract"=>"Trypanosomatid parasites are notorious for the human diseases they cause throughout Africa and South America. However, non-pathogenic trypanosomatids are also found worldwide, infecting a wide range of hosts. One example is Trypanosoma (Megatrypanum) theileri, a ubiquitous protozoan commensal of bovids, which is distributed globally. Exploiting knowledge of pathogenic trypanosomatids, we have developed Trypanosoma theileri as a novel vehicle to deliver vaccine antigens and other proteins to cattle. Conditions for the growth and transfection of T. theileri have been optimised and expressed heterologous proteins targeted for secretion or specific localisation at the cell interior or surface using trafficking signals from Trypanosoma brucei. In cattle, the engineered vehicle could establish in the context of a pre-existing natural T. theileri population, was maintained long-term and generated specific immune responses to an expressed Babesia antigen at protective levels. Building on several decades of basic research into trypanosomatid pathogens, Trypanosoma theileri offers significant potential to target multiple infections, including major cattle-borne zoonoses such as Escherichia coli, Salmonella spp., Brucella abortus and Mycobacterium spp. It also has the potential to deliver therapeutics to cattle, including the lytic factor that protects humans from cattle trypanosomiasis. This could alleviate poverty by protecting indigenous African cattle from African trypanosomiasis.", "link"=>"http://www.mendeley.com/research/targeting-cattleborne-zoonoses-cattle-pathogens-using-novel-trypanosomatidbased-delivery-system", "reader_count"=>64, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>2, "Researcher"=>20, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>16, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>1, "Student > Bachelor"=>8, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>2, "Researcher"=>20, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>16, "Student > Postgraduate"=>1, "Student > Master"=>7, "Other"=>1, "Student > Bachelor"=>8, "Lecturer"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Engineering"=>1, "Environmental Science"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Medicine and Dentistry"=>10, "Agricultural and Biological Sciences"=>32, "Neuroscience"=>1, "Veterinary Science and Veterinary Medicine"=>5, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Immunology and Microbiology"=>5}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>10}, "Neuroscience"=>{"Neuroscience"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>32}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>4}, "Environmental Science"=>{"Environmental Science"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>5}}, "reader_count_by_country"=>{"Brazil"=>1, "United Kingdom"=>2, "Kenya"=>2}, "group_count"=>1}
Scopus 1203 Jan 05:15 UTC

Scopus | Further Information

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Figshare 3419 Feb 04:11 UTC

Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/719068"], "description"=>"<p>(<b>A</b>) An immunofluorescence image of <i>Trypanosoma theileri</i> stained with antibody to <i>T. brucei</i> alpha tubulin; the kinetoplast and nuclear DNA is stained with DAPI (blue). (<b>B</b>) Schematic representation of the integration of an expression cassette into the SSU rRNA locus of the <i>T. theileri</i> genome by homologous recombination. Read-through transcription drives expression from the integrated cassette. (<b>C</b>) Expression vectors used to assay CAT protein expression in <i>T. theileri</i>, this being as an unmodified protein (“Native”) or modified for secretion (“N-BiP”) or surface expression (“N-BiP-GPI”). For the N-BiP and N-BiP-GPI fusions, two gene copies were inserted in tandem to improve overall expression, these being separated by the <i>T. theileri</i> beta-alpha tubulin intergenic region. The respective distribution of CAT protein to the cell pellet or extracellular milieu is indicated on the right hand side, this being quantitated by CAT ELISA assay.</p>", "links"=>[], "tags"=>["proteins", "localized"], "article_id"=>389420, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["G. Adam Mott", "Raymond Wilson", "Anuruddika Fernando", "Ailie Robinson", "Paula MacGregor", "David Kennedy", "Dick Schaap", "Jacqueline B. Matthews", "Keith R. Matthews"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002340.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Heterologous_proteins_can_be_expressed_and_specifically_localized_in_T_theileri_/389420", "title"=>"Heterologous proteins can be expressed and specifically localized in <i>T. theileri</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-10-27 02:37:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/719284"], "description"=>"<p>Northern blots of untransfected <i>T. theileri</i> and <i>T. theileri</i> engineered to express native Bd37, N-BiP Bd37 (‘Bd37 secreted’) or N-BiP Bd37 GPI (‘Bd37 surface’), transcripts being detected using a riboprobe detecting Bd37. Respective loading is indicated by the Ethidium bromide stained rRNA (lower panel).</p>", "links"=>[], "tags"=>["exogenous", "genes", "transgenic"], "article_id"=>389638, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["G. Adam Mott", "Raymond Wilson", "Anuruddika Fernando", "Ailie Robinson", "Paula MacGregor", "David Kennedy", "Dick Schaap", "Jacqueline B. Matthews", "Keith R. Matthews"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002340.g003", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transcript_expression_of_exogenous_genes_in_transgenic_T_theileri_/389638", "title"=>"Transcript expression of exogenous genes in transgenic <i>T. theileri.</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-10-27 02:40:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/719409"], "description"=>"<p>(<b>A</b>) Schematic representation of the amplicon used to detect <i>T. theileri</i> genomic DNA. The amplicon, specific for the <i>T. theileri</i> tubulin gene array, will detect both endogenous <i>T. theileri</i> infection and the presence of the transgenic vaccine vehicle. PCR products were derived from individual calves either before inoculation with transgenic <i>T. theileri</i>, or 1 week and 12 weeks post inoculation. Prior to inoculation 5/17 animals harboured a naturally occurring <i>T. theileri</i> population. (<b>B</b>) A second amplicon specific for the expressed Bd37 gene detects only genomic DNA from the transgenic <i>T. theileri.</i> The cattle all became positive for the transgenic vaccine vehicle within 1 week, regardless of their original infection status, this being sustained throughout the 12 weeks of the experiment for all animals.</p>", "links"=>[], "tags"=>["immunology", "Infectious diseases", "microbiology"], "article_id"=>389762, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["G. Adam Mott", "Raymond Wilson", "Anuruddika Fernando", "Ailie Robinson", "Paula MacGregor", "David Kennedy", "Dick Schaap", "Jacqueline B. Matthews", "Keith R. Matthews"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002340.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inoculation_with_the_vaccine_vehicle_results_in_successful_establishment_of_the_population_in_cattle_/389762", "title"=>"Inoculation with the vaccine vehicle results in successful establishment of the population in cattle.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-10-27 02:42:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/719576"], "description"=>"<p>Northern Blot to determine N-BiP Bd37 transcript expression in cultured <i>T. theileri</i> cells used to inoculate cattle, or harvested after 3 weeks growth in inoculated cattle. Expression of the Bd37 transcript was maintained. Respective loading is indicated by the Ethidium bromide staining rRNA (lower panel).</p>", "links"=>[], "tags"=>["transgene"], "article_id"=>389931, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["G. Adam Mott", "Raymond Wilson", "Anuruddika Fernando", "Ailie Robinson", "Paula MacGregor", "David Kennedy", "Dick Schaap", "Jacqueline B. Matthews", "Keith R. Matthews"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002340.g005", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sustained_transgene_expression_in_T_theileri_after_growth_in_cattle_/389931", "title"=>"Sustained transgene expression in <i>T. theileri</i> after growth in cattle.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-10-27 02:45:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/719659"], "description"=>"<p>(<b>A-C</b>) Quantification of anti-Bd37 antibody responses by ELISA assay demonstrate that calves exposed to Bd37 that is expressed at an internal (<b>A</b>) or surface location (<b>B</b>) on <i>T. theileri</i>, or is secreted (<b>C</b>), show sero-conversion (filled circles) within 12 weeks of inoculation. Animals that did not sero-convert are shown as open circles. (<b>D</b>) End-point titres measured for all experimental animals show that those cattle receiving secreted Bd37 achieved significantly higher antibody levels than the other treatment groups (* p<0.05, ** p<0.001). (<b>E</b>) Time-course of antibody response to secreted Bd37 vaccine vehicle in all cattle. (<b>F</b>) Sera from vaccinated animals are able to compete with an anti-Bd37 monoclonal antibody for ELISA binding; assay details are provided in the ‘<a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002340#s4\" target=\"_blank\">Materials and Methods</a>’. Vertical arrows in Panels C and E indicate inoculation of a further 1×10<sup>6</sup> transgenic <i>T. theileri</i>.</p>", "links"=>[], "tags"=>["stimulates", "antibody"], "article_id"=>390012, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["G. Adam Mott", "Raymond Wilson", "Anuruddika Fernando", "Ailie Robinson", "Paula MacGregor", "David Kennedy", "Dick Schaap", "Jacqueline B. Matthews", "Keith R. Matthews"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002340.g006", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inoculation_with_the_vaccine_vehicle_stimulates_an_effective_antibody_response_to_the_expressed_antigen_/390012", "title"=>"Inoculation with the vaccine vehicle stimulates an effective antibody response to the expressed antigen.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-10-27 00:00:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/719178"], "description"=>"<p>Distribution of expressed Bd37 assayed via its fusion to <i>T. brucei</i> BiP and detected using a BiP-specific antibody. Cells were fixed with paraformaldehyde and then either 0.1% Triton-X100 permeabilised, or not, to distinguish intracellular from surface exposed protein following immunofluorescence. In wild-type cells, only weak staining was observed (<b>A–C</b>) in the absence of permeabilisation whereas permeabilisation (<b>D–F</b>) allows visualisation of endogenous, intracellular <i>T. theileri</i> BiP protein, which cross-reacts with the <i>T. brucei</i> BiP antibody. In contrast, the N-BiP-Bd37 fusion protein generates intense staining at the flagellar-pocket in intact cells (<b>G–I</b>), plus an intense intracellular staining in permeabilised cells comprised of endogenous BiP and N-BiP-Bd37 protein in the ER (<b>J–L</b>). Expression of the N-BiP-Bd37-GPI fusion protein generates a strong signal on intact cells indicating the presence of cell-surface protein (<b>M–O</b>); permeabilised cells also show strong straining representing the combined endogenous BiP and the expressed fusion protein <b>(P–R).</b> Camera exposures with respect to the BiP fluorescence and image handling were precisely consistent between cells under each condition to allow comparison of the relative intensity of signal. DAPI images reveal the kinetoplast and nuclear position, these often overlying one another. Scale Bar = 10 µm.</p>", "links"=>[], "tags"=>["n-bip", "n-bip-gpi", "fusion"], "article_id"=>389529, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["G. Adam Mott", "Raymond Wilson", "Anuruddika Fernando", "Ailie Robinson", "Paula MacGregor", "David Kennedy", "Dick Schaap", "Jacqueline B. Matthews", "Keith R. Matthews"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002340.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cellular_location_of_expressed_N_BiP_and_N_BIP_GPI_fusion_proteins_/389529", "title"=>"Cellular location of expressed N-BiP and N-BIP-GPI fusion proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-10-27 02:38:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/364053"], "description"=>"<div><p>Trypanosomatid parasites are notorious for the human diseases they cause throughout Africa and South America. However, non-pathogenic trypanosomatids are also found worldwide, infecting a wide range of hosts. One example is <em>Trypanosoma (Megatrypanum) theileri</em>, a ubiquitous protozoan commensal of bovids, which is distributed globally. Exploiting knowledge of pathogenic trypanosomatids, we have developed <em>Trypanosoma theileri</em> as a novel vehicle to deliver vaccine antigens and other proteins to cattle. Conditions for the growth and transfection of <em>T. theileri</em> have been optimised and expressed heterologous proteins targeted for secretion or specific localisation at the cell interior or surface using trafficking signals from <em>Trypanosoma brucei</em>. In cattle, the engineered vehicle could establish in the context of a pre-existing natural <em>T. theileri</em> population, was maintained long-term and generated specific immune responses to an expressed <em>Babesia</em> antigen at protective levels. Building on several decades of basic research into trypanosomatid pathogens, <em>Trypanosoma theileri</em> offers significant potential to target multiple infections, including major cattle-borne zoonoses such as <em>Escherichia coli</em>, <em>Salmonella</em> spp., <em>Brucella abortus</em> and <em>Mycobacterium</em> spp. It also has the potential to deliver therapeutics to cattle, including the lytic factor that protects humans from cattle trypanosomiasis. This could alleviate poverty by protecting indigenous African cattle from African trypanosomiasis.</p> </div>", "links"=>[], "tags"=>["targeting", "cattle-borne", "Zoonoses", "pathogens", "trypanosomatid-based"], "article_id"=>131955, "categories"=>["Cancer", "Microbiology", "Immunology"], "users"=>["G. Adam Mott", "Raymond Wilson", "Anuruddika Fernando", "Ailie Robinson", "Paula MacGregor", "David Kennedy", "Dick Schaap", "Jacqueline B. Matthews", "Keith R. Matthews"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002340", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Targeting_Cattle_Borne_Zoonoses_and_Cattle_Pathogens_Using_a_Novel_Trypanosomatid_Based_Delivery_System/131955", "title"=>"Targeting Cattle-Borne Zoonoses and Cattle Pathogens Using a Novel Trypanosomatid-Based Delivery System", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-10-27 00:32:35"}
PMC Usage Stats 124617 Jan 19:46 UTC

PMC Usage Stats | Further Information

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Relative Metric 60448208 Mar 19:39 UTC

Relative Metric

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F1000Prime 120 Jan 13:08 UTC

F1000Prime | Further Information

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