TLR2/MyD88/NF-κB Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection
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{"title"=>"TLR2/MyD88/NF-κB pathway, reactive oxygen species, potassium efflux activates NLRP3/ASC inflammasome during respiratory syncytial virus infection", "type"=>"journal", "authors"=>[{"first_name"=>"Jesus", "last_name"=>"Segovia", "scopus_author_id"=>"50263104000"}, {"first_name"=>"Ahmed", "last_name"=>"Sabbah", "scopus_author_id"=>"25121697300"}, {"first_name"=>"Victoria", "last_name"=>"Mgbemena", "scopus_author_id"=>"35262385500"}, {"first_name"=>"Su Yu", "last_name"=>"Tsai", "scopus_author_id"=>"54925267500"}, {"first_name"=>"Te Hung", "last_name"=>"Chang", "scopus_author_id"=>"7404724789"}, {"first_name"=>"Michael T.", "last_name"=>"Berton", "scopus_author_id"=>"7003481317"}, {"first_name"=>"Ian R.", "last_name"=>"Morris", "scopus_author_id"=>"35321479700"}, {"first_name"=>"Irving C.", "last_name"=>"Allen", "scopus_author_id"=>"12782207700"}, {"first_name"=>"Jenny P Y", "last_name"=>"Ting", "scopus_author_id"=>"7101824874"}, {"first_name"=>"Santanu", "last_name"=>"Bose", "scopus_author_id"=>"7401626489"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"364145941", "sgr"=>"84856249966", "issn"=>"19326203", "pmid"=>"22295065", "scopus"=>"2-s2.0-84856249966", "doi"=>"10.1371/journal.pone.0029695", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"065eccd8-77f2-34fb-9030-ce705934dbf0", "abstract"=>"Human respiratory syncytial virus (RSV) constitute highly pathogenic virus that cause severe respiratory diseases in newborn, children, elderly and immuno-compromised individuals. Airway inflammation is a critical regulator of disease outcome in RSV infected hosts. Although \"controlled\" inflammation is required for virus clearance, aberrant and exaggerated inflammation during RSV infection results in development of inflammatory diseases like pneumonia and bronchiolitis. Interleukin-1β (IL-1β) plays an important role in inflammation by orchestrating the pro-inflammatory response. IL-1β is synthesized as an immature pro-IL-1β form. It is cleaved by activated caspase-1 to yield mature IL-1β that is secreted extracellularly. Activation of caspase-1 is mediated by a multi-protein complex known as the inflammasome. Although RSV infection results in IL-1β release, the mechanism is unknown. Here in, we have characterized the mechanism of IL-1β secretion following RSV infection. Our study revealed that NLRP3/ASC inflammasome activation is crucial for IL-1β production during RSV infection. Further studies illustrated that prior to inflammasome formation; the \"first signal\" constitutes activation of toll-like receptor-2 (TLR2)/MyD88/NF-κB pathway. TLR2/MyD88/NF-κB signaling is required for pro-IL-1β and NLRP3 gene expression during RSV infection. Following expression of these genes, two \"second signals\" are essential for triggering inflammasome activation. Intracellular reactive oxygen species (ROS) and potassium (K(+)) efflux due to stimulation of ATP-sensitive ion channel promote inflammasome activation following RSV infection. Thus, our studies have underscored the requirement of TLR2/MyD88/NF-κB pathway (first signal) and ROS/potassium efflux (second signal) for NLRP3/ASC inflammasome formation, leading to caspase-1 activation and subsequent IL-1β release during RSV infection.", "link"=>"http://www.mendeley.com/research/tlr2myd88nf%CE%BAb-pathway-reactive-oxygen-species-potassium-efflux-activates-nlrp3asc-inflammasome-durin", "reader_count"=>77, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>5, "Researcher"=>12, "Student > Doctoral Student"=>7, "Student > Ph. D. Student"=>26, "Student > Postgraduate"=>7, "Student > Master"=>8, "Other"=>3, "Student > Bachelor"=>5, "Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>5, "Researcher"=>12, "Student > Doctoral Student"=>7, "Student > Ph. D. 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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/350706", "https://ndownloader.figshare.com/files/350729", "https://ndownloader.figshare.com/files/350746", "https://ndownloader.figshare.com/files/350768"], "description"=>"<div><p>Human respiratory syncytial virus (RSV) constitute highly pathogenic virus that cause severe respiratory diseases in newborn, children, elderly and immuno-compromised individuals. Airway inflammation is a critical regulator of disease outcome in RSV infected hosts. Although “controlled” inflammation is required for virus clearance, aberrant and exaggerated inflammation during RSV infection results in development of inflammatory diseases like pneumonia and bronchiolitis. Interleukin-1β (IL-1β) plays an important role in inflammation by orchestrating the pro-inflammatory response. IL-1β is synthesized as an immature pro-IL-1β form. It is cleaved by activated caspase-1 to yield mature IL-1β that is secreted extracellularly. Activation of caspase-1 is mediated by a multi-protein complex known as the inflammasome. Although RSV infection results in IL-1β release, the mechanism is unknown. Here in, we have characterized the mechanism of IL-1β secretion following RSV infection. Our study revealed that NLRP3/ASC inflammasome activation is crucial for IL-1β production during RSV infection. Further studies illustrated that prior to inflammasome formation; the “first signal” constitutes activation of toll-like receptor-2 (TLR2)/MyD88/NF-κB pathway. TLR2/MyD88/NF-κB signaling is required for pro-IL-1β and NLRP3 gene expression during RSV infection. Following expression of these genes, two “second signals” are essential for triggering inflammasome activation. Intracellular reactive oxygen species (ROS) and potassium (K<sup>+</sup>) efflux due to stimulation of ATP-sensitive ion channel promote inflammasome activation following RSV infection. Thus, our studies have underscored the requirement of TLR2/MyD88/NF-κB pathway (first signal) and ROS/potassium efflux (second signal) for NLRP3/ASC inflammasome formation, leading to caspase-1 activation and subsequent IL-1β release during RSV infection.</p> </div>", "links"=>[], "tags"=>["reactive", "potassium", "efflux", "activates", "inflammasome", "respiratory", "syncytial"], "article_id"=>129320, "categories"=>["Cancer", "Microbiology", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.s001", "https://dx.doi.org/10.1371/journal.pone.0029695.s002", "https://dx.doi.org/10.1371/journal.pone.0029695.s003", "https://dx.doi.org/10.1371/journal.pone.0029695.s004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/TLR2_MyD88_NF_B_Pathway_Reactive_Oxygen_Species_Potassium_Efflux_Activates_NLRP3_ASC_Inflammasome_during_Respiratory_Syncytial_Virus_Infection/129320", "title"=>"TLR2/MyD88/NF-κB Pathway, Reactive Oxygen Species, Potassium Efflux Activates NLRP3/ASC Inflammasome during Respiratory Syncytial Virus Infection", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-01-25 02:35:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/688231"], "description"=>"<p>(<b>A</b>) 293 cells were transfected either with pcDNA (control), pro-IL-1β and/or pro-caspase-1 plasmids. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. (<b>B</b>) Human macrophagic U937 cells were infected with RSV (1 MOI) in the presence of either water (vehicle control) or caspase-1 inhibitor (10 µM of Ac-YVAD-CHO). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. (<b>C</b>) Primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of either water (vehicle control) or caspase-1 inhibitor (10 µM of Ac-YVAD-CHO). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. (<b>D</b>) Wild type (WT) or caspase-1 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.</p>", "links"=>[], "tags"=>["rsv"], "article_id"=>358715, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Caspase_1_dependent_IL_1_946_release_during_RSV_infection_/358715", "title"=>"Caspase-1 dependent IL-1β release during RSV infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:25:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/688317"], "description"=>"<p>(<b>A</b>) 293 cells were transfected either with pcDNA (control), pro-IL-1β+pro-caspase-1 plasmids and NLRP3 plasmid. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. (<b>B</b>) Wild type (WT) or NLRP3 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.</p>", "links"=>[], "tags"=>["secretion", "rsv", "requires", "nlrp3"], "article_id"=>358794, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_IL_1_946_secretion_during_RSV_infection_requires_NLRP3_expression_/358794", "title"=>"IL-1β secretion during RSV infection requires NLRP3 expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:26:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/688388"], "description"=>"<p>(<b>A</b>) 293 cells were transfected either with pcDNA (control), pro-IL-1β+pro-caspase-1 plasmids and ASC plasmid. At 24 h post-transfection, cells were infected with RSV (1 MOI). At 12 h post-infection, medium supernatant was assessed for IL-1β protein by ELISA analysis. (<b>B</b>) Wild type (WT) or ASC knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.</p>", "links"=>[], "tags"=>["rsv"], "article_id"=>358871, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_ASC_expression_is_essential_for_IL_1_946_production_during_RSV_infection_/358871", "title"=>"ASC expression is essential for IL-1β production during RSV infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:27:51"}
  • {"files"=>["https://ndownloader.figshare.com/files/688481"], "description"=>"<p>(<b>A</b>) Wild type (WT), NLRP3 knock-out (KO) and ASC KO BMDMs were infected with RSV (1 MOI) for 12 h. The cell lysate from mock infected and RSV infected cells were subjected to Western blot analysis with mouse caspase-1 p10 subunit specific antibody. (<b>B</b>) RT-PCR analysis of pro-IL-1β, ASC and NLRP3 expression in RSV infected WT, NLRP3 KO and ASC KO BMDMs. The gels shown in (A) and (B) is representative of three independent experiments that yielded similar results.</p>", "links"=>[], "tags"=>["asc", "caspase-1", "activation", "rsv"], "article_id"=>358963, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_NLRP3_and_ASC_are_required_for_caspase_1_activation_following_RSV_infection_/358963", "title"=>"NLRP3 and ASC are required for caspase-1 activation following RSV infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:29:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/688562"], "description"=>"<p>(<b>A</b>) Human U937 cells were infected with RSV (1 MOI) in the presence of either DMSO (control) or NF-κB inhibitor (BAY 11-7082). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. (<b>B</b>) RT-PCR analysis of pro-IL-1β, caspase-1 and NLRP3 expression in U937 cells infected with RSV in the presence of either DMSO (control) or BAY 11-7082. (<b>C</b>) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of either DMSO (control) or BAY 11-7082. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. (<b>D</b>) RT-PCR analysis of pro-IL-1β, ASC and NLRP3 expression in WT BMDMs infected with RSV in the presence of either DMSO (control) or BAY 11-7082. The gels shown in (B) and (D) are representative of three independent experiments that yielded similar results. For ELISA results, each value represents the mean ± standard deviation from three independent experiments.</p>", "links"=>[], "tags"=>["signaling", "secretion", "rsv"], "article_id"=>359044, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_NF_954_B_signaling_is_critical_for_IL_1_946_secretion_during_RSV_infection_/359044", "title"=>"NF-κB signaling is critical for IL-1β secretion during RSV infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:30:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/688668"], "description"=>"<p>(<b>A</b>) Wild type (WT), TLR2 knock-out (KO) and TLR4 KO BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. (<b>B</b>) Wild type (WT) and MyD88 knock-out (KO) BMDMs were infected with RSV (1 MOI). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. (<b>C</b>) RT-PCR analysis of pro-IL-1β expression in RSV infected WT, TLR2 KO and MyD88 KO BMDMs. (D) RT-PCR analysis of caspase-1 and NLRP3 expression in RSV infected WT, TLR2 KO and MyD88 KO BMDMs. The gels shown in (C) and (D) are representative of three independent experiments that yielded similar results. For ELISA assay, each value represents the mean ± standard deviation from three independent experiments.</p>", "links"=>[], "tags"=>["pathway", "rsv"], "article_id"=>359152, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TLR2_MyD88_pathway_is_required_for_IL_1_946_release_during_RSV_infection_/359152", "title"=>"TLR2/MyD88 pathway is required for IL-1β release during RSV infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:32:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/688769"], "description"=>"<p>(<b>A</b>) U937 cells were infected with RSV (1 MOI) in the presence of ROS inhibitor APDC (50 µM). Water served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. (<b>B</b>) U937 cells were infected with RSV (1 MOI) in the presence of ROS inhibitor DPI (10 µM). DMSO served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. (<b>C</b>) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of ROS inhibitor DPI (2 µM) or DMSO (vehicle control). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.</p>", "links"=>[], "tags"=>["produced", "rsv", "triggers"], "article_id"=>359250, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_ROS_produced_during_RSV_infection_triggers_IL_1_946_secretion_/359250", "title"=>"ROS produced during RSV infection triggers IL-1β secretion.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:34:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/688837"], "description"=>"<p> (<b>A</b>) Wild type primary mouse bone marrow derived macrophages (BMDM) were infected with RSV (1 MOI) in the presence of ATP-sensitive potassium channel inhibitor glibenclamide (50 µM). DMSO served as the vehicle control. IL-1β levels in the medium supernatant were assayed by ELISA at 12 h and 24 h post-infection. Cells were also treated with LPS and ATP in the presence of either DMSO or glibenclamide. (<b>B</b>) U937 cells were infected with RSV (1 MOI) in the presence of buffer containing either NaCl (150 mM) (control) or KCl (150 mM). IL-1β levels in the medium supernatant were assayed by ELISA at 12 h post-infection. (<b>C</b>) BMDM were infected with RSV (1 MOI) in the presence of buffer containing either NaCl (150 mM) (control) or KCl (150 mM). IL-1β levels in the medium supernatant were assayed by ELISA at 6 h and 12 h post-infection. Each value represents the mean ± standard deviation from three independent experiments.</p>", "links"=>[], "tags"=>["efflux", "rsv"], "article_id"=>359320, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Potassium_efflux_plays_a_critical_role_in_IL_1_946_production_during_RSV_infection_/359320", "title"=>"Potassium efflux plays a critical role in IL-1β production during RSV infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:35:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/688926"], "description"=>"<p>RSV infection activates TLR2/MyD88 pathway that culminates in NF-κB activation. Nuclear translocation of NF-κB results in NF-κB mediated trans-activation of pro-IL-1β and NLRP3 genes. Intracellular ROS generated during RSV infection and potassium efflux due to stimulation of ATP-sensitive ion channel triggers assembly of NLRP3/ASC inflammasome complex. NLRP3/ASC inflammasome activates caspase-1, which subsequently cleaves pro-IL-1β protein into its mature form. Mature IL-1β is secreted from the cells.</p>", "links"=>[], "tags"=>["schematic", "depicting", "secretion", "rsv"], "article_id"=>359404, "categories"=>["Microbiology", "Infectious Diseases", "Immunology"], "users"=>["Jesus Segovia", "Ahmed Sabbah", "Victoria Mgbemena", "Su-Yu Tsai", "Te-Hung Chang", "Michael T. Berton", "Ian R. Morris", "Irving C. Allen", "Jenny P.-Y. Ting", "Santanu Bose"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0029695.g009"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_A_schematic_model_depicting_the_mechanism_of_IL_1_946_secretion_during_RSV_infection_/359404", "title"=>"A schematic model depicting the mechanism of IL-1β secretion during RSV infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-01-25 02:36:44"}

PMC Usage Stats | Further Information

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