Caspase-2 Mediated Apoptotic and Necrotic Murine Macrophage Cell Death Induced by Rough Brucella abortus
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Mendeley | Further Information

{"title"=>"Caspase-2 mediated apoptotic and necrotic murine macrophage cell death induced by rough Brucella abortus", "type"=>"journal", "authors"=>[{"first_name"=>"Fang", "last_name"=>"Chen", "scopus_author_id"=>"57199921587"}, {"first_name"=>"Yongqun", "last_name"=>"He", "scopus_author_id"=>"7404942594"}], "year"=>2009, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"19714247", "doi"=>"10.1371/journal.pone.0006830", "sgr"=>"69949092200", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-69949092200", "issn"=>"19326203", "pui"=>"355212953"}, "id"=>"c70a46a9-819d-3931-8da4-303276daac13", "abstract"=>"Brucella species are Gram-negative, facultative intracellular bacteria that cause zoonotic brucellosis. Survival and replication inside macrophages is critical for establishment of chronic Brucella infection. Virulent smooth B. abortus strain 2308 inhibits programmed macrophage cell death and replicates inside macrophages. Cattle B. abortus vaccine strain RB51 is an attenuated rough, lipopolysaccharide O antigen-deficient mutant derived from smooth strain 2308. B. abortus rough mutant RA1 contains a single wboA gene mutation in strain 2308. Our studies demonstrated that live RB51 and RA1, but not strain 2308 or heat-killed Brucella, induced both apoptotic and necrotic cell death in murine RAW264.7 macrophages and bone marrow derived macrophages. The same phenomenon was also observed in primary mouse peritoneal macrophages from mice immunized intraperitoneally with vaccine strain RB51 using the same dose as regularly performed in protection studies. Programmed macrophage cell death induced by RB51 and RA1 was inhibited by a caspase-2 inhibitor (Z-VDVAD-FMK). Caspase-2 enzyme activation and cleavage were observed at the early infection stage in macrophages infected with RB51 and RA1 but not strain 2308. The inhibition of macrophage cell death promoted the survival of rough Brucella cells inside macrophages. The critical role of caspase-2 in mediating rough B. abortus induced macrophage cell death was confirmed using caspase-2 specific shRNA. The mitochondrial apoptosis pathway was activated in macrophages infected with rough B. abortus as demonstrated by increase in mitochondrial membrane permeability and the release of cytochrome c to cytoplasm in macrophages infected with rough Brucella. These results demonstrate that rough B. abortus strains RB51 and RA1 induce apoptotic and necrotic murine macrophage cell death that is mediated by caspase-2. The biological relevance of Brucella O antigen and caspase-2-mediated macrophage cell death in Brucella pathogenesis and protective Brucella immunity is discussed.", "link"=>"http://www.mendeley.com/research/caspase2-mediated-apoptotic-necrotic-murine-macrophage-cell-death-induced-rough-brucella-abortus", "reader_count"=>29, "reader_count_by_academic_status"=>{"Librarian"=>1, "Researcher"=>4, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>2, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>5, "Lecturer"=>2}, "reader_count_by_user_role"=>{"Librarian"=>1, "Researcher"=>4, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>2, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>5, "Lecturer"=>2}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>2, "Mathematics"=>1, "Agricultural and Biological Sciences"=>17, "Medicine and Dentistry"=>1, "Veterinary Science and Veterinary Medicine"=>1, "Computer Science"=>2, "Immunology and Microbiology"=>4, "Neuroscience"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>17}, "Computer Science"=>{"Computer Science"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Mathematics"=>{"Mathematics"=>1}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"United States"=>1}, "group_count"=>3}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/886452"], "description"=>"<p>(A) Analysis of caspase-2 activities by a colorimetric assay. The asterisk (*) represents significant differences (<i>P</i><0.05) compared to the medium control group. (B) <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006830#s4f\" target=\"_blank\">Western blot analysis</a> of precaspase-2 in infected macrophages. The numbers shown in the figure represent the amounts of procaspase-2 quantified by densitometry and normalized to the β-actin content. The data represent the means±standard deviations from three independent experiments.</p>", "links"=>[], "tags"=>["enzyme", "infected", "macrophages", "induced", "rb51"], "article_id"=>556908, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g007", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Caspase_2_enzyme_activity_in_infected_macrophages_induced_by_RB51_but_not_by_S2308_/556908", "title"=>"Caspase-2 enzyme activity in infected macrophages induced by RB51 but not by S2308.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:53:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/886743"], "description"=>"<p>Infection with S2308 at a MOI of 2,000 did not cause decrease of MMP or cytochrome <i>c</i> release. (A) The results of flow cytometry results were summarized in graphic form, and represent the means±standard deviations of three independent experiments. The asterisk (*) represents significant differences (<i>P</i><0.05) compared to the medium control group. (B) Cytochrome <i>c</i> release was analyzed by <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006830#s4f\" target=\"_blank\">Western blot analysis</a>. β-actin was used as a reference. Only cytoplasmic portion of cell lysates was assayed. The results are a representative of three independent experiments.</p>", "links"=>[], "tags"=>["mitochondrial", "membrane", "cytochrome", "mitochondria", "rb51-infected", "macrophages"], "article_id"=>557193, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g010", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Decreased_mitochondrial_membrane_potential_MMP_and_increased_cytochrome_c_release_from_mitochondria_in_RB51_infected_macrophages_MOI_200_/557193", "title"=>"Decreased mitochondrial membrane potential (MMP) and increased cytochrome <i>c</i> release from mitochondria in RB51-infected macrophages (MOI: 200).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:55:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/886531"], "description"=>"<p>Macrophages pretreated with Z-VDVAD-FMK were infected with RB51 for 24 h at a MOI of 200. (A) Macrophages pretreated with the caspase-2 inhibitor and infected with RB51 released less LDH than infected cells lacking inhibitor. (B) Growth kinetics of RB51 in macrophages. The number of RB51 cells surviving inside macrophages pretreated with the caspase-2 inhibitor was approximately 120 times greater than the number of cells found in untreated macrophages (<i>P</i><0.05). The asterisk (*) represents significant difference (<i>P</i><0.05) in the LDH release or bacterial CFUs from RB51-infected macrophages with caspase-2 inhibitor treatment compared to that without caspase-2 inhibitor. Data represent the means±standard deviations from three independent experiments.</p>", "links"=>[], "tags"=>["infected", "macrophages", "rb51", "usage", "caspase-2"], "article_id"=>556986, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g008", "stats"=>{"downloads"=>2, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Decreased_cell_death_of_infected_macrophages_and_increased_RB51_survival_inside_macrophages_with_the_usage_of_the_caspase_2_inhibitor_/556986", "title"=>"Decreased cell death of infected macrophages and increased RB51 survival inside macrophages with the usage of the caspase-2 inhibitor.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:53:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/886354"], "description"=>"<p>Macrophages were pretreated with individual caspase inhibitor and then infected with RB51 or RA1 at a MOI of 200. No inhibition was observed with any of the other caspase inhibitor. The results are a representative of three independent experiments.</p>", "links"=>[], "tags"=>["macrophage", "pan-caspase", "inhibitor", "caspase-2"], "article_id"=>556808, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g006", "stats"=>{"downloads"=>2, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inhibition_of_rough_B_abortus_induced_macrophage_cell_death_by_a_pan_caspase_inhibitor_and_a_caspase_2_inhibitor_/556808", "title"=>"Inhibition of rough <i>B. abortus</i>-induced macrophage cell death by a pan-caspase inhibitor and a caspase-2 inhibitor.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:52:57"}
  • {"files"=>["https://ndownloader.figshare.com/files/886185"], "description"=>"<p>Annexin V/PI staining shows both apoptosis and necrosis of peritoneal macrophages at 24 h after infection with RB51. No obvious cell death was observed in peritoneal macrophages from mice injected (i.p.) with S2308 or heat-killed RB51 (HK-RB51). The results are a representative of three independent experiments.</p>", "links"=>[], "tags"=>["necrotic", "peritoneal", "macrophages", "rb51-vaccinated"], "article_id"=>556642, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g005", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Apoptotic_and_necrotic_cell_death_in_peritoneal_primary_macrophages_isolated_from_RB51_vaccinated_mice_/556642", "title"=>"Apoptotic and necrotic cell death in peritoneal primary macrophages isolated from RB51-vaccinated mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:52:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/886079"], "description"=>"<p>Smooth strain S2308 (MOI: 2000) and heat-killed RB51 and RA1 (equivalent MOI: 200) did not induce obvious macrophage cell death. The results are a representative of three independent experiments.</p>", "links"=>[], "tags"=>["necrotic", "marrow", "derived", "macrophages", "infected", "viable", "strains", "rb51", "ra1", "24"], "article_id"=>556528, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g004", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Apoptotic_and_necrotic_cell_death_of_bone_marrow_derived_macrophages_infected_with_viable_rough_B_abortus_strains_RB51_or_RA1_MOI_200_at_24_h_post_infection_/556528", "title"=>"Apoptotic and necrotic cell death of bone marrow derived macrophages infected with viable rough <i>B. abortus</i> strains RB51 or RA1 (MOI: 200) at 24 h post infection.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:51:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/886627"], "description"=>"<p>Macrophages with or without caspase-2 shRNA were infected for 6 h with RB51 at a MOI of 200. (A) Caspase-2 expression in different clones was detected by <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0006830#s4f\" target=\"_blank\">Western blot analysis</a>. The numbers shown in the figure represent the amounts of procaspase-2 that were quantified by densitometry and normalized to the β-actin data. (B) The LDH assay was used to detect differential cell death in different macrophage clones. The asterisk (*) represents significant differences (<i>P</i><0.05) of LDH releases from different clones compared to that from the parent RAW264.7 cells. The data represent the means±standard deviations from three independent experiments.</p>", "links"=>[], "tags"=>["rb51-induced", "macrophage", "caspase-2"], "article_id"=>557085, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g009", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inhibition_of_RB51_induced_macrophage_cell_death_by_caspase_2_shRNA_/557085", "title"=>"Inhibition of RB51-induced macrophage cell death by caspase-2 shRNA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:54:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/885957"], "description"=>"<p>(A) Uneven macrophage internalization of RB51GFP with different infection MOIs observed at 1 h post infection. (B) Most cell death occurred in macrophages infected with RB51GFP (MOI: 200) at 6 h post infection. The results are a representative of three independent experiments.</p>", "links"=>[], "tags"=>["internalization", "macrophage"], "article_id"=>556418, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g003", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_the_relationship_between_internalization_of_rough_Brucella_and_macrophage_cell_death_/556418", "title"=>"Analysis of the relationship between internalization of rough <i>Brucella</i> and macrophage cell death.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:50:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/886803"], "description"=>"a<p>Hours Post Infection.</p>b<p>Results of Annexin V and/or PI staining of macrophages infected with indicated bacteria: ++++, 75 to 100% positive; +++, 50 to 75% positive; ++, 25 to 50% positive; +, 5–25% positive; ±, <5% positive; -, no positive staining.</p>", "links"=>[], "tags"=>["macrophage"], "article_id"=>557272, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.t001", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Kinetic_analysis_of_Brucella_induced_macrophage_cell_death_/557272", "title"=>"Kinetic analysis of <i>Brucella</i>-induced macrophage cell death.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-21 04:55:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/885831"], "description"=>"<p>(A) Annexin V/PI staining shows both apoptosis (solid arrow) and necrosis (hollow arrow) of macrophages at as early as 2 h post infection. (B) Staining with Hoechst 33342 shows cell shrinkage and nuclear condensation (diamond head arrow) of infected macrophages at 6 h post infection at a MOI of 200. The results are a representative of three independent experiments.</p>", "links"=>[], "tags"=>["necrotic", "macrophage", "induced"], "article_id"=>556292, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g002", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Apoptotic_and_necrotic_macrophage_cell_death_induced_by_RB51_/556292", "title"=>"Apoptotic and necrotic macrophage cell death induced by RB51.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:50:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/885735"], "description"=>"<p>(A) RB51 and RA1 induced 61.5%±8.7% and 74.7%±5.6% macrophage cells death, respectively, at a MOI of 200 at 24 h post infection. <i>B. abortus</i> strain 2308 induced limited macrophage cell death even at the MOI of 2,000. (B) LDH release from RB51, RA1 and S2308 infected macrophages. The data represent the means±standard deviations for three independent experiments. The asterisk (*) represents the significant differences (<i>P</i><0.05) of the LDH release level from macrophages infected by smooth S2308 compared to that from macrophages infected with rough strain RB51 or RA1. (C) Growth kinetics of RB51, RA1 and S2308 in macrophages. The number of internalized cells of S2308 at MOI 2,000 found was similar to the number of cells of internalized RB51 and RA1 at a MOI 200 at 1 h post infection. The asterisk (*) represents the significant differences (<i>P</i><0.05) in survival level of smooth S2308 compared to that of rough strain RB51 or RA1 inside infected macrophages.</p>", "links"=>[], "tags"=>["macrophages", "infected", "strains"], "article_id"=>556191, "categories"=>["Immunology", "Microbiology", "Infectious Diseases"], "users"=>["Fang Chen", "Yongqun He"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0006830.g001", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cell_death_of_macrophages_infected_with_B_abortus_strains_RB51_RA1_and_their_parent_wild_type_S2308_/556191", "title"=>"Cell death of macrophages infected with <i>B. abortus</i> strains RB51,RA1 and their parent wild type S2308.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 04:49:42"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"10", "full-text"=>"7", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"6", "full-text"=>"3", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}

Relative Metric

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