The Lipopolysaccharide Core of Brucella abortus Acts as a Shield Against Innate Immunity Recognition
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{"title"=>"The lipopolysaccharide core of Brucella abortus acts as a shield against innate immunity recognition", "type"=>"journal", "authors"=>[{"first_name"=>"Raquel", "last_name"=>"Conde-Álvarez", "scopus_author_id"=>"24741005300"}, {"first_name"=>"Vilma", "last_name"=>"Arce-Gorvel", "scopus_author_id"=>"24741059800"}, {"first_name"=>"Maite", "last_name"=>"Iriarte", "scopus_author_id"=>"7006715517"}, {"first_name"=>"Mateja", "last_name"=>"Manček-Keber", "scopus_author_id"=>"14527522000"}, {"first_name"=>"Elías", "last_name"=>"Barquero-Calvo", "scopus_author_id"=>"22133768300"}, {"first_name"=>"Leyre", "last_name"=>"Palacios-Chaves", "scopus_author_id"=>"36873307600"}, {"first_name"=>"Carlos", "last_name"=>"Chacón-Díaz", "scopus_author_id"=>"23569410500"}, {"first_name"=>"Esteban", "last_name"=>"Chaves-Olarte", "scopus_author_id"=>"6602745898"}, {"first_name"=>"Anna", "last_name"=>"Martirosyan", "scopus_author_id"=>"7003381218"}, {"first_name"=>"Kristine", "last_name"=>"von Bargen", "scopus_author_id"=>"23669713000"}, {"first_name"=>"María Jesús", "last_name"=>"Grilló", "scopus_author_id"=>"57188637123"}, {"first_name"=>"Roman", "last_name"=>"Jerala", "scopus_author_id"=>"7004624486"}, {"first_name"=>"Klaus", "last_name"=>"Brandenburg", "scopus_author_id"=>"7007147548"}, {"first_name"=>"Enrique", "last_name"=>"Llobet", "scopus_author_id"=>"6603269819"}, {"first_name"=>"José A.", "last_name"=>"Bengoechea", "scopus_author_id"=>"6603841760"}, {"first_name"=>"Edgardo", "last_name"=>"Moreno", "scopus_author_id"=>"7202827718"}, {"first_name"=>"Ignacio", "last_name"=>"Moriyón", "scopus_author_id"=>"7004333011"}, {"first_name"=>"Jean Pierre", "last_name"=>"Gorvel", "scopus_author_id"=>"7005675644"}], "year"=>2012, "source"=>"PLoS Pathogens", "identifiers"=>{"doi"=>"10.1371/journal.ppat.1002675", "sgr"=>"84874554030", "issn"=>"15537366", "pui"=>"365220862", "isbn"=>"1553-7374 (Electronic)\\r1553-7366 (Linking)", "pmid"=>"22589715", "scopus"=>"2-s2.0-84874554030"}, "id"=>"3a211074-9872-3dc6-aa63-a06366380b3a", "abstract"=>"Innate immunity recognizes bacterial molecules bearing pathogen-associated molecular patterns to launch inflammatory responses leading to the activation of adaptive immunity. However, the lipopolysaccharide (LPS) of the gram-negative bacterium Brucella lacks a marked pathogen-associated molecular pattern, and it has been postulated that this delays the development of immunity, creating a gap that is critical for the bacterium to reach the intracellular replicative niche. We found that a B. abortus mutant in the wadC gene displayed a disrupted LPS core while keeping both the LPS O-polysaccharide and lipid A. In mice, the wadC mutant induced proinflammatory responses and was attenuated. In addition, it was sensitive to killing by non-immune serum and bactericidal peptides and did not multiply in dendritic cells being targeted to lysosomal compartments. In contrast to wild type B. abortus, the wadC mutant induced dendritic cell maturation and secretion of pro-inflammatory cytokines. All these properties were reproduced by the wadC mutant purified LPS in a TLR4-dependent manner. Moreover, the core-mutated LPS displayed an increased binding to MD-2, the TLR4 co-receptor leading to subsequent increase in intracellular signaling. Here we show that Brucella escapes recognition in early stages of infection by expressing a shield against recognition by innate immunity in its LPS core and identify a novel virulence mechanism in intracellular pathogenic gram-negative bacteria. These results also encourage for an improvement in the generation of novel bacterial vaccines.", "link"=>"http://www.mendeley.com/research/lipopolysaccharide-core-brucella-abortus-acts-shield-against-innate-immunity-recognition", "reader_count"=>106, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Librarian"=>4, "Researcher"=>19, "Student > Doctoral Student"=>8, "Student > Ph. D. Student"=>29, "Student > Postgraduate"=>3, "Student > Master"=>13, "Other"=>6, "Student > Bachelor"=>12, "Lecturer"=>1, "Professor"=>7}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Librarian"=>4, "Researcher"=>19, "Student > Doctoral Student"=>8, "Student > Ph. D. 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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/640400"], "description"=>"<p>Western blot analyses were performed with monoclonal antibodies Cby-33H8 (O-polysaccharide C/Y epitope) and A68/24D08/G09 (core oligosaccharide) and SDS-proteinase K LPS extracts of Ba-parental, BaΔ<i>wadC</i> and BaΔ<i>wadC</i>-compl (complemented mutant), and the SDS-PAGE electropherogram of LPS phenol-water extracts was silver stained.</p>", "links"=>[], "tags"=>["carries", "defective", "lps"], "article_id"=>310872, "categories"=>["Biological Sciences"], "users"=>["Raquel Conde-Álvarez", "Vilma Arce-Gorvel", "Maite Iriarte", "Mateja Manček-Keber", "Elías Barquero-Calvo", "Leyre Palacios-Chaves", "Carlos Chacón-Díaz", "Esteban Chaves-Olarte", "Anna Martirosyan", "Kristine von Bargen", "María-Jesús Grilló", "Roman Jerala", "Klaus Brandenburg", "Enrique Llobet", "José A. Bengoechea", "Edgardo Moreno", "Ignacio Moriyón", "Jean-Pierre Gorvel"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002675.g001", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ba_916_wadC_carries_a_partially_defective_LPS_core_oligosaccharide_/310872", "title"=>"Ba<i>ΔwadC</i> carries a partially defective LPS core oligosaccharide.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:14:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/330633", "https://ndownloader.figshare.com/files/330687", "https://ndownloader.figshare.com/files/330726", "https://ndownloader.figshare.com/files/330769", "https://ndownloader.figshare.com/files/330871", "https://ndownloader.figshare.com/files/330912", "https://ndownloader.figshare.com/files/330996", "https://ndownloader.figshare.com/files/331069", "https://ndownloader.figshare.com/files/331095", "https://ndownloader.figshare.com/files/331145", "https://ndownloader.figshare.com/files/331184"], "description"=>"<div><p>Innate immunity recognizes bacterial molecules bearing pathogen-associated molecular patterns to launch inflammatory responses leading to the activation of adaptive immunity. However, the lipopolysaccharide (LPS) of the gram-negative bacterium <em>Brucella</em> lacks a marked pathogen-associated molecular pattern, and it has been postulated that this delays the development of immunity, creating a gap that is critical for the bacterium to reach the intracellular replicative niche. We found that a <em>B. abortus</em> mutant in the <em>wadC</em> gene displayed a disrupted LPS core while keeping both the LPS O-polysaccharide and lipid A. In mice, the <em>wadC</em> mutant induced proinflammatory responses and was attenuated. In addition, it was sensitive to killing by non-immune serum and bactericidal peptides and did not multiply in dendritic cells being targeted to lysosomal compartments. In contrast to wild type <em>B. abortus</em>, the <em>wadC</em> mutant induced dendritic cell maturation and secretion of pro-inflammatory cytokines. All these properties were reproduced by the <em>wadC</em> mutant purified LPS in a TLR4-dependent manner. Moreover, the core-mutated LPS displayed an increased binding to MD-2, the TLR4 co-receptor leading to subsequent increase in intracellular signaling. Here we show that <em>Brucella</em> escapes recognition in early stages of infection by expressing a shield against recognition by innate immunity in its LPS core and identify a novel virulence mechanism in intracellular pathogenic gram-negative bacteria. These results also encourage for an improvement in the generation of novel bacterial vaccines.</p> </div>", "links"=>[], "tags"=>["lipopolysaccharide", "acts", "shield", "innate", "immunity"], "article_id"=>125288, "categories"=>["Biological Sciences"], "users"=>["Raquel Conde-Álvarez", "Vilma Arce-Gorvel", "Maite Iriarte", "Mateja Manček-Keber", "Elías Barquero-Calvo", "Leyre Palacios-Chaves", "Carlos Chacón-Díaz", "Esteban Chaves-Olarte", "Anna Martirosyan", "Kristine von Bargen", "María-Jesús Grilló", "Roman Jerala", "Klaus Brandenburg", "Enrique Llobet", "José A. Bengoechea", "Edgardo Moreno", "Ignacio Moriyón", "Jean-Pierre Gorvel"], "doi"=>["https://dx.doi.org/10.1371/journal.ppat.1002675.s001", "https://dx.doi.org/10.1371/journal.ppat.1002675.s002", "https://dx.doi.org/10.1371/journal.ppat.1002675.s003", "https://dx.doi.org/10.1371/journal.ppat.1002675.s004", "https://dx.doi.org/10.1371/journal.ppat.1002675.s005", "https://dx.doi.org/10.1371/journal.ppat.1002675.s006", "https://dx.doi.org/10.1371/journal.ppat.1002675.s007", "https://dx.doi.org/10.1371/journal.ppat.1002675.s008", "https://dx.doi.org/10.1371/journal.ppat.1002675.s009", "https://dx.doi.org/10.1371/journal.ppat.1002675.s010", "https://dx.doi.org/10.1371/journal.ppat.1002675.s011"], "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/The_Lipopolysaccharide_Core_of_Brucella_abortus_Acts_as_a_Shield_Against_Innate_Immunity_Recognition/125288", "title"=>"The Lipopolysaccharide Core of <em>Brucella abortus</em> Acts as a Shield Against Innate Immunity Recognition", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-05-10 01:28:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/640660"], "description"=>"<p><b>Upper panel</b>, intracellular replication of Ba-parental, BaΔ<i>wadC</i>, BaΔ<i>wadC</i>-compl and <i>virB</i> mutant in BMDC and BMDM. Each point is the mean ± standard error of an experiment performed in triplicate, and the results representative of three independent experiments. Differences between Ba-parental and BaΔ<i>wadC</i> are indicated with symbols (#, p<0.001). <b>Lower panel</b>, confocal imaging of Ba-parental and BaΔ<i>wadC</i> carrying a GFP plasmid at 24 h post-infection (calnexin and LAMP-1 are in red).</p>", "links"=>[], "tags"=>["attenuated"], "article_id"=>311132, "categories"=>["Biological Sciences"], "users"=>["Raquel Conde-Álvarez", "Vilma Arce-Gorvel", "Maite Iriarte", "Mateja Manček-Keber", "Elías Barquero-Calvo", "Leyre Palacios-Chaves", "Carlos Chacón-Díaz", "Esteban Chaves-Olarte", "Anna Martirosyan", "Kristine von Bargen", "María-Jesús Grilló", "Roman Jerala", "Klaus Brandenburg", "Enrique Llobet", "José A. Bengoechea", "Edgardo Moreno", "Ignacio Moriyón", "Jean-Pierre Gorvel"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002675.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ba_wadC_is_attenuated_in_BMDC_/311132", "title"=>"BaΔ<i>wadC</i> is attenuated in BMDC.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:18:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/640767"], "description"=>"<p>(<b>A</b>), IL-12 p40/p70 and TNF-α released by BMDC 24 h after infection (left panel) or stimulation with 10 µg/mL of Ba-parental or BaΔ<i>wadC</i> LPS, or with 100 ng/mL of <i>E. coli</i> LPS (right panel) measured by ELISA; (<b>B</b>), IL-12 p40/p70 (left panel) and TNF-α (right panel) released by BMDC from wild-type (WT), TLR9−/− TLR4−/− or TLR2−/− mice 24 h after stimulation with 10 µg/mL of Ba-parental or BaΔ<i>wadC</i> LPS, or with 100 ng/mL of <i>E. coli</i> LPS measured by ELISA; (<b>C</b>), Surface expression of CD40, CD86 and MHCII in LPS-stimulated BMDC measured by flow cytometry. Each point is the mean ± standard error (n = 3). But for TLR4−/− mice differences between Ba-parental and BaΔ<i>wadC</i> or their corresponding LPS were statistically significant (p<0.001). Results are representative of two independent experiments.</p>", "links"=>[], "tags"=>["induces", "cytokine", "reproduced", "purified", "lps", "tlr-4"], "article_id"=>311229, "categories"=>["Biological Sciences"], "users"=>["Raquel Conde-Álvarez", "Vilma Arce-Gorvel", "Maite Iriarte", "Mateja Manček-Keber", "Elías Barquero-Calvo", "Leyre Palacios-Chaves", "Carlos Chacón-Díaz", "Esteban Chaves-Olarte", "Anna Martirosyan", "Kristine von Bargen", "María-Jesús Grilló", "Roman Jerala", "Klaus Brandenburg", "Enrique Llobet", "José A. Bengoechea", "Edgardo Moreno", "Ignacio Moriyón", "Jean-Pierre Gorvel"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002675.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ba_wadC_induces_a_cytokine_release_that_is_reproduced_by_purified_LPS_in_a_TLR_4_dependent_fashion_/311229", "title"=>"BaΔ<i>wadC</i> induces a cytokine release that is reproduced by purified LPS in a TLR-4 dependent fashion.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:20:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/640490"], "description"=>"<p>(<b>A</b>), Left panel, infection kinetics in the spleens of mice intraperitoneally inoculated with 5×10<sup>4</sup> Ba-parental, BaΔ<i>wadC</i> or 1×10<sup>8</sup> BaTn5::<i>per</i>; right panel, spleen weights. (<b>B</b>), Leukocyte and PMN levels in the peritoneum (left panel) and blood (right panel) of mice after intraperitoneal infection with 1×10<sup>6</sup> Ba-parental, BaΔ<i>wadC</i> or 1×10<sup>5 </sup><i>S.</i> Typhimurium SL1344. (<b>C</b>), TNF-α, IL-6, IL-10 and IL-12 p40/p70 levels in the sera of mice infected with Ba-parental or BaΔ<i>wadC</i>. Each point is the mean ± standard deviation (n = 5). Differences between Ba-parental and BaΔ<i>wadC</i> are indicated with symbols (*, p<0.05; #, p<0.001).</p>", "links"=>[], "tags"=>["attenuated", "mice", "induces", "transitory", "leukocyte", "recruitment", "cytokine"], "article_id"=>310956, "categories"=>["Biological Sciences"], "users"=>["Raquel Conde-Álvarez", "Vilma Arce-Gorvel", "Maite Iriarte", "Mateja Manček-Keber", "Elías Barquero-Calvo", "Leyre Palacios-Chaves", "Carlos Chacón-Díaz", "Esteban Chaves-Olarte", "Anna Martirosyan", "Kristine von Bargen", "María-Jesús Grilló", "Roman Jerala", "Klaus Brandenburg", "Enrique Llobet", "José A. Bengoechea", "Edgardo Moreno", "Ignacio Moriyón", "Jean-Pierre Gorvel"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002675.g002", "stats"=>{"downloads"=>1, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ba_wadC_is_attenuated_in_mice_and_induces_transitory_splenomegaly_local_leukocyte_recruitment_and_cytokine_secretion_/310956", "title"=>"BaΔ<i>wadC</i> is attenuated in mice and induces transitory splenomegaly, local leukocyte recruitment and cytokine secretion.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:15:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/640906"], "description"=>"<p>(<b>A</b>), Survival of Ba-parental, BaΔ<i>wadC</i> and BaTn5::<i>per</i> after incubation in non-immune calf serum for 90 min. Each point is the mean ± standard error of an experiment performed in triplicate, and the results representative of five independent experiments (differences between Ba-parental and BaΔ<i>wadC</i> are indicated as #, p<0.001). (<b>B</b>), minimal inhibitory concentrations (MIC) of polymyxin B (determined by the serial dilution method) and colistin (determined by the E-test) (results representative of three independent experiments). (<b>C</b>), Crystalline to fluid (β↔α) phase transition of the hydrocarbon chains of Ba-parental LPS and BaΔ<i>wadC</i> LPS measured in the presence (filled circles) or absence (empty circles) of normal human serum (red circles) or polymyxin B (black circles; LPS∶PMB 1∶0.1 molar ratio). The plots represent the position (wavenumber) of the peak of the symmetric stretching vibration of the methylene groups νs(CH2) versus temperature. Tc, transition temperature (results are representative of three independent experiments).</p>", "links"=>[], "tags"=>["shows", "serum", "bactericidal", "peptides", "relates", "lps"], "article_id"=>311373, "categories"=>["Biological Sciences"], "users"=>["Raquel Conde-Álvarez", "Vilma Arce-Gorvel", "Maite Iriarte", "Mateja Manček-Keber", "Elías Barquero-Calvo", "Leyre Palacios-Chaves", "Carlos Chacón-Díaz", "Esteban Chaves-Olarte", "Anna Martirosyan", "Kristine von Bargen", "María-Jesús Grilló", "Roman Jerala", "Klaus Brandenburg", "Enrique Llobet", "José A. Bengoechea", "Edgardo Moreno", "Ignacio Moriyón", "Jean-Pierre Gorvel"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002675.g005", "stats"=>{"downloads"=>1, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ba_wadC_shows_increased_sensitivity_to_normal_serum_and_bactericidal_peptides_that_relates_to_the_LPS_core_defect_/311373", "title"=>"BaΔ<i>wadC</i> shows increased sensitivity to normal serum and bactericidal peptides that relates to the LPS core defect.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:22:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/641046"], "description"=>"<p>(<b>A</b>) LPS binding to hMD-2. After incubation of 0.75 µM hMD-2 with increasing LPS concentrations, the fraction of hMD-2 not bound to LPS was detected with free-hMD-2 specific antibody 9B4 by ELISA (results are representative of three independent experiments run in triplicate; differences between Ba-parental and BaΔ<i>wadC</i> are indicated as *, p<0.05 or #, p<0.001) (<b>B</b>) NF-κB translocation to the nucleus. BMDC were stimulated for 1 h with media, <i>E. coli</i> LPS (100 ng/mL), Ba-parental LPS (10 µg/mL) or BaΔ<i>wadC</i> LPS (10 µg/mL), cells fixed in 3% paraformaldehyde at 37°C for 15 min, immunostained for CD11c and MHCII, and the % of cells showing nuclear translocation of the NF-κB subunit p65/RelA recorded (results are representative of three independent experiments run in triplicate; differences between Ba-parental and BaΔ<i>wadC</i> are indicated as #, p<0.001). (<b>C</b>) S6 phosphorylation. BMDC were stimulated for 30 min, 1, 4, or 6 h with (left to right) media, <i>E. coli</i> LPS (100 ng/mL), Ba-parental LPS (100 ng/mL), Ba-parental LPS (10 µg/mL) or BaΔ<i>wadC</i> LPS (10 µg/mL). Cell lysates (30 µg protein) were analyzed by Western blot with a polyclonal antibody against S6-P or an anti-actin antibody as control.</p>", "links"=>[], "tags"=>["lps", "binds", "md-2", "increases", "translocation", "s6"], "article_id"=>311501, "categories"=>["Biological Sciences"], "users"=>["Raquel Conde-Álvarez", "Vilma Arce-Gorvel", "Maite Iriarte", "Mateja Manček-Keber", "Elías Barquero-Calvo", "Leyre Palacios-Chaves", "Carlos Chacón-Díaz", "Esteban Chaves-Olarte", "Anna Martirosyan", "Kristine von Bargen", "María-Jesús Grilló", "Roman Jerala", "Klaus Brandenburg", "Enrique Llobet", "José A. Bengoechea", "Edgardo Moreno", "Ignacio Moriyón", "Jean-Pierre Gorvel"], "doi"=>"https://dx.doi.org/10.1371/journal.ppat.1002675.g006", "stats"=>{"downloads"=>4, "page_views"=>23, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Ba_wadC_LPS_binds_to_MD_2_and_increases_NF_B_translocation_and_S6_phosphorylation_/311501", "title"=>"BaΔ<i>wadC</i> LPS binds to MD-2 and increases NF-κB translocation and S6 phosphorylation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-05-10 00:25:01"}

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