Transmission-Blocking Antibodies against Mosquito C-Type Lectins for Dengue Prevention
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{"title"=>"Transmission-Blocking Antibodies against Mosquito C-Type Lectins for Dengue Prevention", "type"=>"journal", "authors"=>[{"first_name"=>"Yang", "last_name"=>"Liu", "scopus_author_id"=>"56040362000"}, {"first_name"=>"Fuchun", "last_name"=>"Zhang", "scopus_author_id"=>"57189252300"}, {"first_name"=>"Jianying", "last_name"=>"Liu", "scopus_author_id"=>"56060556600"}, {"first_name"=>"Xiaoping", "last_name"=>"Xiao", "scopus_author_id"=>"56060434400"}, {"first_name"=>"Siyin", "last_name"=>"Zhang", "scopus_author_id"=>"56060543700"}, {"first_name"=>"Chengfeng", "last_name"=>"Qin", "scopus_author_id"=>"16304943900"}, {"first_name"=>"Ye", "last_name"=>"Xiang", "scopus_author_id"=>"36112393500"}, {"first_name"=>"Penghua", "last_name"=>"Wang", "scopus_author_id"=>"23483045700"}, {"first_name"=>"Gong", "last_name"=>"Cheng", "scopus_author_id"=>"36672614000"}], "year"=>2014, "source"=>"PLoS Pathogens", "identifiers"=>{"sgr"=>"84895742280", "doi"=>"10.1371/journal.ppat.1003931", "pui"=>"372548643", "pmid"=>"24550728", "scopus"=>"2-s2.0-84895742280", "issn"=>"15537374", "isbn"=>"10.1371/journal.ppat.1003931"}, "id"=>"d279a4fa-fe57-34a9-93cc-caf753bac9f5", "abstract"=>"C-type lectins are a family of proteins with carbohydrate-binding activity. Several C-type lectins in mammals or arthropods are employed as receptors or attachment factors to facilitate flavivirus invasion. We previously identified a C-type lectin in Aedes aegypti, designated as mosquito galactose specific C-type lectin-1 (mosGCTL-1), facilitating the attachment of West Nile virus (WNV) on the cell membrane. Here, we first identified that 9 A. aegypti mosGCTL genes were key susceptibility factors facilitating DENV-2 infection, of which mosGCTL-3 exhibited the most significant effect. We found that mosGCTL-3 was induced in mosquito tissues with DENV-2 infection, and that the protein interacted with DENV-2 surface envelop (E) protein and virions in vitro and in vivo. In addition, the other identified mosGCTLs interacted with the DENV-2 E protein, indicating that DENV may employ multiple mosGCTLs as ligands to promote the infection of vectors. The vectorial susceptibility factors that facilitate pathogen invasion may potentially be explored as a target to disrupt the acquisition of microbes from the vertebrate host. Indeed, membrane blood feeding of antisera against mosGCTLs dramatically reduced mosquito infective ratio. Hence, the immunization against mosGCTLs is a feasible approach for preventing dengue infection. Our study provides a future avenue for developing a transmission-blocking vaccine that interrupts the life cycle of dengue virus and reduces disease burden.", "link"=>"http://www.mendeley.com/research/transmissionblocking-antibodies-against-mosquito-ctype-lectins-dengue-prevention", "reader_count"=>65, "reader_count_by_academic_status"=>{"Researcher"=>14, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>23, "Student > Postgraduate"=>4, "Student > Master"=>8, "Other"=>2, "Student > Bachelor"=>11, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Researcher"=>14, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>23, "Student > Postgraduate"=>4, "Student > Master"=>8, "Other"=>2, "Student > Bachelor"=>11, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>10, "Agricultural and Biological Sciences"=>37, "Medicine and Dentistry"=>7, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>2, "Psychology"=>2, "Social Sciences"=>1, "Immunology and Microbiology"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>7}, "Chemistry"=>{"Chemistry"=>2}, "Social Sciences"=>{"Social Sciences"=>1}, "Psychology"=>{"Psychology"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>37}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>10}, "Unspecified"=>{"Unspecified"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"Brazil"=>1, "United Kingdom"=>2, "Portugal"=>1}, "group_count"=>7}

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  • {"files"=>["https://ndownloader.figshare.com/files/1385843"], "description"=>"<p>(<b>A–F</b>) Two low-passage DENV-2 strains, which were isolated from patients' sera, were employed to test transmission-blocking effect of mosGCTL antisera in <i>A. aegypti</i>. The pre-immune sera, mosGCTL-3 antisera and combined mosGCTLs antisera (1/9 mosGCTL antiserum each) with Vero cells-generatedDENV-2 AF2014178 strain (A to C)/JX470186 strain (D to F) and human blood were used for mosquitoes feeding. The antisera were serially diluted 100- (A and D), 1,000- (B and E) and 5,000- (C and F) fold for the investigation. The fed mosquitoes were sacrificed on 8 days post infection to determine the infectivity by qPCR. The numbers of infected mosquitoes/total mosquitoes are presented on the top of each column (Left panel of each sub-figure). The mosquito infective ratio (Right panel) indicates the ratio of DENV positive to total mosquitoes. One dot represents a mosquito. The results are pooled from 3 independent experiments.</p>", "links"=>[], "tags"=>["microbiology", "Emerging infectious diseases", "Host-pathogen interaction", "Vector biology", "Virology", "mosgctls", "antisera", "low-passage", "denv-2"], "article_id"=>934573, "categories"=>["Biological Sciences"], "users"=>["Yang Liu", "Fuchun Zhang", "Jianying Liu", "Xiaoping Xiao", "Siyin Zhang", "Chengfeng Qin", "Ye Xiang", "Penghua Wang", "Gong Cheng"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003931.g006", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transmission_blocking_effect_of_mosGCTLs_antisera_in_the_infection_of_low_passage_DENV_2_strains_/934573", "title"=>"Transmission-blocking effect of mosGCTLs antisera in the infection of low-passage DENV-2 strains.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-13 03:00:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/1385824"], "description"=>"<p>(<b>A</b>) mosGCTL-3 interacted with DENV-2 Envelop protein in a co-immunoprecipitation (Co-IP) assay. The purified S2-expressed mosGCTL-3 and DENV-2 E proteins (2 ug each) were mixed for 2 h at 4°C. The protein complex was pulled down by anti-V5 mouse monoclonal antibody and probed by anti-FLAG rabbit polyclonal antibody. The experiment was repeated twice. (<b>B</b>) mosGCTL-3 captured DENV-2 virions by ELISA. The purified S2-expressed mosGCTL-3 was used in this experiment. The interaction was determined by a flavivirus E mAb 4G2. Data was expressed as mean ± standard error, and the experiment was reproduced three times. (<b>C–D)</b> The co-localization of mosGCTL-3 and DENV-2 in the tissues of <i>A. aegypti</i>. The mosquitoes were infected by intrathoracic microinjection. The salivary glands and hemocytes were then collected on 6 days after 1,000 M.I.D.<sub>50</sub> DENV-2 infection for immunofluorescence staining. The tissues of PBS inoculated mosquitoes were used as mocks. mosGCTL-3 was stained with anti-rabbit IgG Alexa-488 (Green), and DENV-2 E protein was probed by anti-mouse IgG Alexa-546 (Red). Nuclei were stained blue with To-Pro-3 iodide. The arrow represented the associated area between mosGCTL-3 and DENV-2 in hemocytes (C) and salivary glands (D). Images were examined by a Zeiss LSM 780 meta confocal with a Multi-Track mode.</p>", "links"=>[], "tags"=>["microbiology", "Emerging infectious diseases", "Host-pathogen interaction", "Vector biology", "Virology", "mosgctl-3"], "article_id"=>934562, "categories"=>["Biological Sciences"], "users"=>["Yang Liu", "Fuchun Zhang", "Jianying Liu", "Xiaoping Xiao", "Siyin Zhang", "Chengfeng Qin", "Ye Xiang", "Penghua Wang", "Gong Cheng"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003931.g003", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_interaction_between_mosGCTL_3_and_DENV_2_/934562", "title"=>"The interaction between mosGCTL-3 and DENV-2.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-13 03:00:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/1385818"], "description"=>"<p>(<b>A–B</b>) Silencing efficiency of <i>mosGCTL-3</i> on both RNA level and protein level. 2 ug <i>GFP</i> or <i>mosGCTL-3</i> dsRNA was inoculated into mosquitoes thorax respectively. At 3, 6 and 9 days after the treatment, the mosquitoes were sacrificed to determine silencing effect by qPCR and normalized by <i>A. aegypti actin</i> (A). The primers of dsRNA synthesis and qPCR detection were described in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003931#ppat.1003931.s008\" target=\"_blank\">Table S3</a>. The <i>mosGCTL-3</i> silenced or mock mosquitoes were grinded in lysis buffer and mosGCTL-3 was detected by immunoblotting (B). (<b>C–F</b>) <i>mosGCTL-3</i> suppression impairs the infection of dengue serotypes. 10 M.I.D.<sub>50</sub> DENV-1 Hawaii strain (C), DENV-2 New Guinea C strain (D), DENV-3 Guangdong strain (E) and DENV-4 H241 strain (F) viruses were inoculated into <i>mosGCTL-3</i> silenced mosquitoes by microinjection respectively. The viral load was determined at 6 days post-infection by qPCR and normalized by <i>A. aegypti actin</i>. The result was pooled from 3 independent experiments. (<b>G</b>) Diagram of <i>mosGCTL-3</i> gene. An extra exon (Exon 1) of <i>mosGCTL-3</i>, which encoded a peptide with 20 amino acid signal sequence, was identified by the 5′-RACE. (<b>H</b>) Expression and purification of mosGCTL-3 by a <i>Drosophila</i> expression system. <i>mosGCTL-3</i> gene was cloned into pMT/BiP/V5-His-A DNA vector. The recombinant mosGCTL-3 was expressed and purified by a Cobalt-His column (Left panel). The expression was probed by anti-V5 mAb (Right panel). Mock was the concentrated supernatant of empty vector transfected <i>Drosophila</i> S2 cells. (<b>I</b>) Inoculation of mosGCTL-3 purified protein benefited DENV-2 infection in <i>A. aegypti</i>. 50 pg or 500 pg purified S2-expressed mosGCTL-3 protein was microinjected into each mosquito with 10 M.I.D.<sub>50</sub> DENV-2. The infected mosquitoes were sacrificed at 3 and 6 days post infection. The same amount of BSA was inoculated with DENV-2 as mocks. The viral load was determined by qPCR and normalized by <i>A. aegypti actin</i>. This experiment was repeated 3 times individually. One dot represented 1 mosquito and the horizontal line represented the mean value in all figures. We used the <i>Mann-Whitney</i> test for statistical analysis.</p>", "links"=>[], "tags"=>["microbiology", "Emerging infectious diseases", "Host-pathogen interaction", "Vector biology", "Virology", "facilitated", "denv"], "article_id"=>934556, "categories"=>["Biological Sciences"], "users"=>["Yang Liu", "Fuchun Zhang", "Jianying Liu", "Xiaoping Xiao", "Siyin Zhang", "Chengfeng Qin", "Ye Xiang", "Penghua Wang", "Gong Cheng"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003931.g002", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_mosGCTL_3_facilitated_DENV_infections_/934556", "title"=>"<i>mosGCTL-3</i> facilitated DENV infections.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-13 03:00:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/1385815"], "description"=>"<p>The phenotype, of which the 9 <i>mosGCTL</i> genes exhibited susceptible effect in DENV-2 (New Guinea C strain) infection (<i>p<0.05</i>, <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003931#ppat.1003931.s006\" target=\"_blank\">Table S1</a>), was reproduced. There were no less than 15 mosquitoes in each group. The quantification of <i>DENV-2 E</i> gene was normalized with <i>A. aegypti actin</i> (<i>AAEL011197</i>). The primers of dsRNA synthesis were shown in <a href=\"http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003931#ppat.1003931.s008\" target=\"_blank\">Table S3</a>. All data were presented as mean ± standard error(SEM. The <i>Mann-Whitney</i> test was used for statistical analysis. The results were combined from 2 independent experiments.</p>", "links"=>[], "tags"=>["microbiology", "Emerging infectious diseases", "Host-pathogen interaction", "Vector biology", "Virology", "genes", "denv-2"], "article_id"=>934553, "categories"=>["Biological Sciences"], "users"=>["Yang Liu", "Fuchun Zhang", "Jianying Liu", "Xiaoping Xiao", "Siyin Zhang", "Chengfeng Qin", "Ye Xiang", "Penghua Wang", "Gong Cheng"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003931.g001", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_role_of_mosGCTL_genes_in_DENV_2_infection_of_A_aegypti_/934553", "title"=>"The role of <i>mosGCTL</i> genes in DENV-2 infection of <i>A. aegypti</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-13 03:00:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/1385832"], "description"=>"<p>(<b>A</b>) The interaction of the susceptible mosGCTLs and DENV-2 E protein. The <i>mosGCTL</i> genes exhibiting susceptible role in DENV-2 infection (<i>p<0.05</i>) were cloned into pMT/Bip/V5-His A DNA vector. The recombinant mosGCTLs with a V5 tag in C-terminus were expressed in <i>Drosophila</i> S2 cells. mosGCTL-26 (<i>AAEL017265</i>) was failed to be expressed in <i>Drosophila</i> S2 cells. The interaction was determined by ELISA. An anti-V5 mouse mAb was used to probe the binding activity. The experiment was reproduced twice. (<b>B–D</b>) Feeding the combination of 9 mosGCTL antisera blocked DENV-2 infection. The pre-immune sera, mosGCTL-3 antisera and combined mosGCTLs antisera (1/9 mosGCTL antiserum each) were serially diluted 100- (B), 1,000- (C), or 5,000- (D) fold with fresh human blood and Vero cells-generated DENV-2. The fed mosquitoes were sacrificed on 8 days post infection to determine the infectivity by qPCR. The number of infected mosquitoes/total mosquitoes was presented on the top of each column (Left panel of each subfigure). The mosquito infective ratio (Right panel) was then interpreted from the left panel. One dot represented a mosquito. The result was pooled from 4 independent experiments.</p>", "links"=>[], "tags"=>["microbiology", "Emerging infectious diseases", "Host-pathogen interaction", "Vector biology", "Virology", "mosgctls", "antisera", "blocked", "denv-2", "uptake", "membrane"], "article_id"=>934570, "categories"=>["Biological Sciences"], "users"=>["Yang Liu", "Fuchun Zhang", "Jianying Liu", "Xiaoping Xiao", "Siyin Zhang", "Chengfeng Qin", "Ye Xiang", "Penghua Wang", "Gong Cheng"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003931.g005", "stats"=>{"downloads"=>0, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Feeding_the_combination_of_9_mosGCTLs_antisera_blocked_DENV_2_uptake_by_a_membrane_blood_feeding_/934570", "title"=>"Feeding the combination of 9 mosGCTLs antisera blocked DENV-2 uptake by a membrane blood feeding.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-13 03:00:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/1385827"], "description"=>"<p>(<b>A</b>) Diagram of dengue life cycle and a transmission-blocking strategy for dengue prevention. (<b>B</b>) Inoculation of mosGCTL-3 antisera impaired DENV-2 infection of <i>A. aegypti</i>. The serial dilutions of mosGCTL-3 antisera or pre-immune sera with 10 M.I.D.<sub>50</sub> DENV-2 were inoculated into mosquitoes by microinjection. The infected mosquitoes were scarified at 3 days and 6 days post inoculation. DENV-2 was determined by qPCR and normalized by <i>A. aegypti actin</i>. One dot represented 1 mosquito and the horizontal line represented the mean value in all figures. The <i>Mann-Whitney</i> test was used for statistical analysis. The result was combined from 2 independent experiments. (<b>C–D</b>) mosGCTL-3 antisera interrupted DENV-2 infection by a blood meal. The mosGCTL-3 antisera or pre-immune sera were diluted 100-, 1,000- and 5,000-fold with fresh human blood and Vero cells-generated DENV-2. The mosquitoes were allowed to ingest the blood mixture by a membrane feeding. The fed mosquitoes were sacrificed 8 days later and DENV-2 infectivity was assessed by qPCR. The number of infected mosquitoes/total mosquitoes was presented on the top of each column (C). The mosquito infective ratio (D) was then interpreted from the Figure (C). One dot represented a mosquito. The result was pooled from 2 independent experiments.</p>", "links"=>[], "tags"=>["microbiology", "Emerging infectious diseases", "Host-pathogen interaction", "Vector biology", "Virology", "antisera", "interrupted", "denv-2"], "article_id"=>934565, "categories"=>["Biological Sciences"], "users"=>["Yang Liu", "Fuchun Zhang", "Jianying Liu", "Xiaoping Xiao", "Siyin Zhang", "Chengfeng Qin", "Ye Xiang", "Penghua Wang", "Gong Cheng"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1003931.g004", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_mosGCTL_3_antisera_interrupted_DENV_2_infection_of_A_aegypti_/934565", "title"=>"mosGCTL-3 antisera interrupted DENV-2 infection of <i>A. aegypti</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-13 03:00:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/1385848", "https://ndownloader.figshare.com/files/1385849", "https://ndownloader.figshare.com/files/1385850", "https://ndownloader.figshare.com/files/1385851", "https://ndownloader.figshare.com/files/1385852", "https://ndownloader.figshare.com/files/1385853", "https://ndownloader.figshare.com/files/1385854", "https://ndownloader.figshare.com/files/1385855"], "description"=>"<div><p>C-type lectins are a family of proteins with carbohydrate-binding activity. Several C-type lectins in mammals or arthropods are employed as receptors or attachment factors to facilitate flavivirus invasion. We previously identified a C-type lectin in <i>Aedes aegypti</i>, designated as mosquito galactose specific C-type lectin-1 (<i>mosGCTL-1</i>), facilitating the attachment of West Nile virus (WNV) on the cell membrane. Here, we first identified that 9 <i>A. aegypti mosGCTL</i> genes were key susceptibility factors facilitating DENV-2 infection, of which <i>mosGCTL-3</i> exhibited the most significant effect. We found that <i>mosGCTL-3</i> was induced in mosquito tissues with DENV-2 infection, and that the protein interacted with DENV-2 surface envelop (E) protein and virions <i>in vitro</i> and <i>in vivo</i>. In addition, the other identified mosGCTLs interacted with the DENV-2 E protein, indicating that DENV may employ multiple mosGCTLs as ligands to promote the infection of vectors. The vectorial susceptibility factors that facilitate pathogen invasion may potentially be explored as a target to disrupt the acquisition of microbes from the vertebrate host. Indeed, membrane blood feeding of antisera against mosGCTLs dramatically reduced mosquito infective ratio. Hence, the immunization against mosGCTLs is a feasible approach for preventing dengue infection. Our study provides a future avenue for developing a transmission-blocking vaccine that interrupts the life cycle of dengue virus and reduces disease burden.</p></div>", "links"=>[], "tags"=>["microbiology", "Emerging infectious diseases", "Host-pathogen interaction", "Vector biology", "Virology", "antibodies", "mosquito", "c-type", "lectins", "dengue"], "article_id"=>934578, "categories"=>["Biological Sciences"], "users"=>["Yang Liu", "Fuchun Zhang", "Jianying Liu", "Xiaoping Xiao", "Siyin Zhang", "Chengfeng Qin", "Ye Xiang", "Penghua Wang", "Gong Cheng"], "doi"=>["https://dx.doi.org/10.1371/journal.ppat.1003931.s001", "https://dx.doi.org/10.1371/journal.ppat.1003931.s002", "https://dx.doi.org/10.1371/journal.ppat.1003931.s003", "https://dx.doi.org/10.1371/journal.ppat.1003931.s004", "https://dx.doi.org/10.1371/journal.ppat.1003931.s005", "https://dx.doi.org/10.1371/journal.ppat.1003931.s006", "https://dx.doi.org/10.1371/journal.ppat.1003931.s007", "https://dx.doi.org/10.1371/journal.ppat.1003931.s008"], "stats"=>{"downloads"=>15, "page_views"=>25, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transmission_Blocking_Antibodies_against_Mosquito_C_Type_Lectins_for_Dengue_Prevention_/934578", "title"=>"Transmission-Blocking Antibodies against Mosquito C-Type Lectins for Dengue Prevention", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-02-13 03:00:48"}

PMC Usage Stats | Further Information

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