Ligation of Macrophage Fcγ Receptors Recapitulates the Gene Expression Pattern of Vulnerable Human Carotid Plaques
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{"title"=>"Ligation of macrophage FCγ receptors recapitulates the gene expression pattern of vulnerable human carotid plaques", "type"=>"journal", "authors"=>[{"first_name"=>"Michelle R.", "last_name"=>"Lennartz", "scopus_author_id"=>"35615577300"}, {"first_name"=>"Ankur", "last_name"=>"Aggarwal", "scopus_author_id"=>"46760952600"}, {"first_name"=>"Tanya M.", "last_name"=>"Michaud", "scopus_author_id"=>"12142252800"}, {"first_name"=>"Paul J.", "last_name"=>"Feustel", "scopus_author_id"=>"7003309156"}, {"first_name"=>"David M.", "last_name"=>"Jones", "scopus_author_id"=>"56216006000"}, {"first_name"=>"M. Julia", "last_name"=>"Brosnan", "scopus_author_id"=>"7005806402"}, {"first_name"=>"Rebecca S.", "last_name"=>"Keller", "scopus_author_id"=>"7401434890"}, {"first_name"=>"Daniel J.", "last_name"=>"Loegering", "scopus_author_id"=>"7006682246"}, {"first_name"=>"Paul B.", "last_name"=>"Kreienberg", "scopus_author_id"=>"7004487982"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203", "pmid"=>"21814555", "doi"=>"10.1371/journal.pone.0021803", "pui"=>"362190924", "issn"=>"19326203", "sgr"=>"79960629247", "scopus"=>"2-s2.0-79960629247"}, "id"=>"33e2a00f-e10c-3359-8c51-111c45dcb64e", "abstract"=>"Stroke is a leading cause of death in the United States. As ∼60% of strokes result from carotid plaque rupture, elucidating the mechanisms that underlie vulnerability is critical for therapeutic intervention. We tested the hypothesis that stable and vulnerable human plaques differentially express genes associated with matrix degradation. Examination established that femoral, and the distal region of carotid, plaques were histologically stable while the proximal carotid plaque regions were vulnerable. Quantitative RT-PCR was used to compare expression of 22 genes among these tissues. Distal carotid and femoral gene expression was not significantly different, permitting the distal carotid segments to be used as a paired control for their corresponding proximal regions. Analysis of the paired plaques revealed differences in 16 genes that impact plaque stability: matrix metalloproteinases (MMP, higher in vulnerable), MMP modulators (inhibitors: lower, activators: higher in vulnerable), activating Fc receptors (FcγR, higher in vulnerable) and FcγR signaling molecules (higher in vulnerable). Surprisingly, the relative expression of smooth muscle cell and macrophage markers in the three plaque types was not significantly different, suggesting that macrophage distribution and/or activation state correlates with (in)stability. Immunohistochemistry revealed that macrophages and smooth muscle cells localize to distinct and non-overlapping regions in all plaques. MMP protein localized to macrophage-rich regions. In vitro, treatment of macrophages with immune complexes, but not oxidized low density lipoprotein, C-reactive protein, or TNF-α, induced a gene expression profile similar to that of the vulnerable plaques. That ligation of FcγR recapitulates the pattern of gene expression in vulnerable plaques suggests that the FcγR → macrophage activation pathway may play a greater role in human plaque vulnerability than previously appreciated.", "link"=>"http://www.mendeley.com/research/ligation-macrophage-fc%CE%B3-receptors-recapitulates-gene-expression-pattern-vulnerable-human-carotid-pla", "reader_count"=>7, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>2, "Other"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>3, "Student > Ph. D. Student"=>2, "Other"=>1}, "reader_count_by_subject_area"=>{"Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Environmental Science"=>{"Environmental Science"=>1}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/753822"], "description"=>"<p><b>(A)</b> Carotid endarterectomy specimen with proximal (P) and distal (D) regions indicated (bar  = 1 cm). Tri-chrome staining of the proximal (top, left) and distal (top, right) regions reveals necrotic cores with a thin cap compromised by cellular infiltration (bottom, left) or protected by a thick fibrous cap (bottom, right), respectively. Bars  = 1 mm. (B) Femoral plaques are characterized by an organized matrix (M) overlying a fibrocellular (FC) band deposited on a calcified (Ca) deposit. The lumen is designated with an L. Bar  = 1 mm.</p>", "links"=>[], "tags"=>["carotid", "femoral"], "article_id"=>424187, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Histology_of_human_carotid_and_femoral_plaques_/424187", "title"=>"Histology of human carotid and femoral plaques.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:09:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/754754"], "description"=>"<p>Differentiated THP-1 cells or monocyte-derived macrophages were treated with 200 ng/ml human TNF-α for 18 h. RNA was extracted and subjected to qPCR. Data are presented as mean ± SEM (normalized to β-actin; n indicated in control column). Statistical significance compared by paired t-test for each replicate. * p<0.05, ** p<.01, *** p<.001. Yellow boxes denote genes similarly regulated in proximal vs distal plaque tissue. Green and red numbers indicate significantly increased or decreased expression, respectively compared to controls.<sup> = </sup> Significance calculated from log values to compensate for heteroscedacity of the data.</p>", "links"=>[], "tags"=>["induce"], "article_id"=>425125, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g008", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_TNF_945_does_not_induce_a_vulnerable_gene_profile_in_macrophages_/425125", "title"=>"TNF-α does not induce a vulnerable gene profile in macrophages.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:25:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/754128"], "description"=>"<p>The proximal (P) and distal (D) regions of a carotid plaque were stained with Trichrome to visualize fibrous tissue (blue) and cellular material (red). Smooth muscle cells and macrophages were visualized by immunohistochemistry for α-actin (smooth muscle cells) and CD68 (macrophages). Smooth muscle cells co-localize with the fibrous material while macrophages localize to acellular areas.</p>", "links"=>[], "tags"=>["cells", "localize", "discrete", "regions", "proximal", "distal", "carotid"], "article_id"=>424487, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Macrophages_and_smooth_muscle_cells_localize_to_discrete_regions_in_both_the_proximal_and_distal_regions_of_carotid_plaques_/424487", "title"=>"Macrophages and smooth muscle cells localize to discrete regions in both the proximal and distal regions of carotid plaques.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:14:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/379333", "https://ndownloader.figshare.com/files/379384"], "description"=>"<div><p>Stroke is a leading cause of death in the United States. As ∼60% of strokes result from carotid plaque rupture, elucidating the mechanisms that underlie vulnerability is critical for therapeutic intervention. We tested the hypothesis that stable and vulnerable human plaques differentially express genes associated with matrix degradation. Examination established that femoral, and the distal region of carotid, plaques were histologically stable while the proximal carotid plaque regions were vulnerable. Quantitative RT-PCR was used to compare expression of 22 genes among these tissues. Distal carotid and femoral gene expression was not significantly different, permitting the distal carotid segments to be used as a paired control for their corresponding proximal regions. Analysis of the paired plaques revealed differences in 16 genes that impact plaque stability: matrix metalloproteinases (MMP, higher in vulnerable), MMP modulators (inhibitors: lower, activators: higher in vulnerable), activating Fc receptors (FcγR, higher in vulnerable) and FcγR signaling molecules (higher in vulnerable). Surprisingly, the relative expression of smooth muscle cell and macrophage markers in the three plaque types was not significantly different, suggesting that macrophage distribution and/or activation state correlates with (in)stability. Immunohistochemistry revealed that macrophages and smooth muscle cells localize to distinct and non-overlapping regions in all plaques. MMP protein localized to macrophage-rich regions. <em>In vitro</em>, treatment of macrophages with immune complexes, but not oxidized low density lipoprotein, C-reactive protein, or TNF-α, induced a gene expression profile similar to that of the vulnerable plaques. That ligation of FcγR recapitulates the pattern of gene expression in vulnerable plaques suggests that the FcγR → macrophage activation pathway may play a greater role in human plaque vulnerability than previously appreciated.</p> </div>", "links"=>[], "tags"=>["ligation", "macrophage", "receptors", "recapitulates", "carotid", "plaques"], "article_id"=>134952, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0021803.s001", "https://dx.doi.org/10.1371/journal.pone.0021803.s002"], "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Ligation_of_Macrophage_Fc_Receptors_Recapitulates_the_Gene_Expression_Pattern_of_Vulnerable_Human_Carotid_Plaques/134952", "title"=>"Ligation of Macrophage Fcγ Receptors Recapitulates the Gene Expression Pattern of Vulnerable Human Carotid Plaques", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-07-21 01:22:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/754950"], "description"=>"<p>*Data are presented as mean ± SD (%).</p><p>$ Statistically Significant.</p>", "links"=>[], "tags"=>["characteristics"], "article_id"=>425322, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.t001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_General_characteristics_of_patient_pool_/425322", "title"=>"General characteristics of patient pool.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-07-21 01:28:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/754613"], "description"=>"<p>7 µm cryosections of carotid plaques were stained for macrophages (CD68) or smooth muscle cells (α-actin). Sequential sections were stained for the indicated TIMPs as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021803#s2\" target=\"_blank\">Methods</a>. Isotype controls are presented in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021803#pone-0021803-g004\" target=\"_blank\">Figure 4</a>. TIMPs 1 and 3 co-localized with macrophages; TIMP2 was expressed in both macrophages and smooth muscle cells. Representative of three plaques giving similar results.</p>", "links"=>[], "tags"=>["inhibitor", "metalloproteinases", "proximal", "carotid", "plaque"], "article_id"=>424975, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g007", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Localization_of_tissue_inhibitor_of_metalloproteinases_TIMPs_in_proximal_carotid_plaque_tissue_/424975", "title"=>"Localization of tissue inhibitor of metalloproteinases (TIMPs) in proximal carotid plaque tissue.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:22:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/754357"], "description"=>"<p>7 µm cryosections were stained for macrophages (CD68) or smooth muscle cells (α-actin) by immunohistochemistry. CD68 and α-actin localized to discrete regions of the plaque. Sequential sections were stained for the indicated MMPs. MMP 8, 9, and 12 localized predominantly to the macrophages; MMP1 was present in both smooth muscle cells and macrophages. L marks plaque lumen. Representative of three plaques giving similar results.</p>", "links"=>[], "tags"=>["metalloproteinases", "localize", "macrophages", "proximal", "regions", "carotid"], "article_id"=>424720, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g005", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Matrix_metalloproteinases_localize_to_macrophages_in_proximal_regions_of_carotid_plaques_/424720", "title"=>"Matrix metalloproteinases localize to macrophages in proximal regions of carotid plaques.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:18:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/754864"], "description"=>"<p>Differentiated THP-1 cells or monocyte-derived macrophages were treated with immune complexes (10 beads/cell) or oxidized LDL (50 µg/ml). RNA was extracted and subjected to qPCR. Data are presented as mean ± SEM (normalized to β-actin; n indicated in control column). Statistical significance compared by paired t-test for each replicate. * p<0.05, ** p<.01, *** p<.001. Yellow boxes denote genes similarly regulated in proximal vs distal plaque tissue. Green and red numbers indicate significantly increased or decreased expression, respectively compared to controls. <sup> = </sup> Significance calculated from log values to compensate for heteroscedacity of the data.</p>", "links"=>[], "tags"=>["complexes", "recapitulate"], "article_id"=>425234, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g009", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immune_complexes_recapitulate_the_gene_expression_pattern_of_vulnerable_plaques_/425234", "title"=>"Immune complexes recapitulate the gene expression pattern of vulnerable plaques.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:27:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/754526"], "description"=>"<p>Distal (D) and proximal (P) regions were isolated from three carotid plaques and MMP9 levels were assessed by zymography using 3 µg of protein from each sample lysate. Proximal regions contained more MMP-9 protein of both the pro and active forms, paralleling the mRNA expression (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021803#pone-0021803-g002\" target=\"_blank\">Figure 2</a>) and consistent with regulation of MMP-9 expression at the transcriptional level. +:20 µg of HT15 media was loaded as a positive control <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021803#pone.0021803-Ranganathan1\" target=\"_blank\">[10]</a>.</p>", "links"=>[], "tags"=>["regions", "carotid", "plaques", "mmp-9", "corresponding", "distal"], "article_id"=>424883, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g006", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Proximal_regions_of_carotid_plaques_contain_more_MMP_9_than_corresponding_distal_regions_/424883", "title"=>"Proximal regions of carotid plaques contain more MMP-9 than corresponding distal regions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:21:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/753951"], "description"=>"<p>Distal carotid plaque tissue and femoral tissue have similar gene expression patterns; proximal and distal carotid tissue have significantly different levels of these genes. The relative mRNA expression for each gene for each plaque is represented by a single point. For each gene and tissue, a Box plot is shown to the left of the actual data. The Box plot indicates the median by a horizontal line within the box; the top and bottom of the box represent the 75<sup>th</sup> and 25<sup>th</sup> percentile, respectively; the whiskers indicate the range of the data and the dot indicates the mean. Statistical significance (p<0.05) is indicated by an asterisk (*) for proximal vs. distal and by a dagger (†) for femoral vs. distal. Expression was compared for femoral and distal carotid specimens using an unpaired Student's t-test.</p>", "links"=>[], "tags"=>["levels", "genes", "implicated", "carotid", "plaque"], "article_id"=>424315, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g002", "stats"=>{"downloads"=>3, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_levels_for_genes_implicated_in_carotid_plaque_vulnerability_/424315", "title"=>"Expression levels for genes implicated in carotid plaque vulnerability.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:11:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/754238"], "description"=>"<p>A proximal/distal ratio was calculated from the qRT-PCR data generated for each paired sample in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021803#pone-0021803-g002\" target=\"_blank\">Figure 2</a>. Results are presented as mean ± SEM (n = 8–16). Genes with a ratio >1 are more highly expressed in proximal tissue; ratios <1 indicate a decrease in gene expression in the proximal vs distal regions. Genes are grouped by color according to function. Statistical analysis and number of samples are presented in the accompanying table. Only genes that were significantly different (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0021803#pone-0021803-g002\" target=\"_blank\">Figure 2</a>) are presented.</p>", "links"=>[], "tags"=>["proximal", "distal", "plaque"], "article_id"=>424605, "categories"=>["Immunology"], "users"=>["Michelle R. Lennartz", "Ankur Aggarwal", "Tanya M. Michaud", "Paul J. Feustel", "David M. Jones", "M. Julia Brosnan", "Rebecca S. Keller", "Daniel J. Loegering", "Paul B. Kreienberg"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0021803.g004", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Relative_gene_expression_in_proximal_vs_distal_plaque_regions_/424605", "title"=>"Relative gene expression in proximal vs distal plaque regions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-21 01:16:45"}

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