Vaccination Targeting a Surface Sialidase of P. acnes: Implication for New Treatment of Acne Vulgaris
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{"title"=>"Vaccination targeting a surface sialidase of P. acnes: Implication for new treatment of acne vulgaris", "type"=>"journal", "authors"=>[{"first_name"=>"Teruaki", "last_name"=>"Nakatsuji", "scopus_author_id"=>"23101032800"}, {"first_name"=>"Yu Tsueng", "last_name"=>"Liu", "scopus_author_id"=>"36071758200"}, {"first_name"=>"Cheng Po", "last_name"=>"Huang", "scopus_author_id"=>"35083564200"}, {"first_name"=>"Richard L.", "last_name"=>"Gallo", "scopus_author_id"=>"7201433211"}, {"first_name"=>"Chun Ming", "last_name"=>"Huang", "scopus_author_id"=>"55731250300"}], "year"=>2008, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-45249113958", "sgr"=>"45249113958", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0001551", "pmid"=>"18253498", "isbn"=>"1932-6203", "pui"=>"351836836"}, "id"=>"a8d5f4fb-431e-318f-9870-fd90035dc629", "abstract"=>"BackgroundAcne vulgaris afflicts more than fifty million people in the United State and the severity of this disorder is associated with the immune response to Propionibacterium acnes (P. acnes). Systemic therapies for acne target P. acnes using antibiotics, or target the follicle with retinoids such as isotretinoin. The latter systemic treatment is highly effective but also carries a risk of side effects including immune imbalance, hyperlipidemia, and teratogenicity. Despite substantial research into potential new therapies for this common disease, vaccines against acne vulgaris are not yet available.Methods and FindingsHere we create an acne vaccine targeting a cell wall-anchored sialidase of P. acnes. The importance of sialidase to disease pathogenesis is shown by treatment of a human sebocyte cell line with recombinant sialidase that increased susceptibility to P. acnes cytotoxicity and adhesion. Mice immunized with sialidase elicit a detectable antibody; the anti-sialidase serum effectively neutralized the cytotoxicity of P. acnes in vitro and P. acnes-induced interleukin-8 (IL-8) production in human sebocytes. Furthermore, the sialidase-immunized mice provided protective immunity against P. acnes in vivo as this treatment blocked an increase in ear thickness and release of pro-inflammatory macrophage inflammatory protein (MIP-2) cytokine.ConclusionsResults indicated that acne vaccines open novel therapeutic avenues for acne vulgaris and other P. acnes-associated diseases.", "link"=>"http://www.mendeley.com/research/vaccination-targeting-surface-sialidase-p-acnes-implication-new-treatment-acne-vulgaris", "reader_count"=>41, "reader_count_by_academic_status"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Librarian"=>2, "Researcher"=>14, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>3, "Other"=>5, "Student > Master"=>5, "Student > Bachelor"=>3, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>3, "Professor > Associate Professor"=>1, "Librarian"=>2, "Researcher"=>14, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>2, "Student > Postgraduate"=>3, "Other"=>5, "Student > Master"=>5, "Student > Bachelor"=>3, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>3, "Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>19, "Medicine and Dentistry"=>9, "Chemistry"=>2, "Social Sciences"=>1, "Immunology and Microbiology"=>5}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>9}, "Chemistry"=>{"Chemistry"=>2}, "Social Sciences"=>{"Social Sciences"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>19}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}, "Unspecified"=>{"Unspecified"=>3}}, "reader_count_by_country"=>{"Colombia"=>2, "United States"=>1, "United Kingdom"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/938065"], "description"=>"<p>(A) Sialic acids on the cell surface of immortalized human sebocytes (SZ95) were detected by their reaction with biotinylated MAA lectin I (10 µg/ml) and streptavidin-FITC conjugate. FITC-fluorescence intensity was measured by flow cytomtery (FACSCalibur, BD Biosciences, San Jose, CA), reflecting the quantity of sialic acid. The sebocytes were pre-treated with 10 µg/ml of purified recombinant sialidase (green, a), GFP (green, b) or an equal volume of PBS (vehicle) (purple) at pH 6 for 2 h. The decrease in FITC-fluorescence intensity in sialidase-treated sebocytes validated the enzymatic activity of the purified sialidase. (B) After pre-treatment with sialidase, sebocytes were co-cultured with <i>P. acnes</i> (5×10<sup>7</sup> CFU/5×10<sup>6</sup> cells) for 18 h. <i>P. acnes</i>-induced cell death in PBS (vehicle)-, sialidase- or GFP-pretreated sebocytes was assessed by trypan blue. Cell death is presented as the % of dead cells compared to all cultured cells. (C) After washing out unbound <i>P. acnes</i>, the number of <i>P. acnes</i> adhered to sebocytes was calculated by spreading Triton-X (0.01%) lysed sebocytes on agar plates to quantify CFU/cell. (D) The CFU/cell in vehicle-pretreated sebocytes was defined as 100%. Adherence of <i>P. acnes</i> (arrows) to vehicle (a)-, sialidase (b)- or GFP (c)-treated sebocytes was visualized by staining with the Accustain Gram stain kit. Pre-treatment with sialidase significantly increased the adhesion of <i>P. acnes</i> to sebocytes. Shown are representative results of three independent experiments. Error bars are mean±SE (<i>P</i><0.01*, <i>P</i><0.001**, <i>P</i><0.0005*** by Student's t-test). Bar: 10 µm.</p>", "links"=>[], "tags"=>["sialic", "acids", "sebocytes", "sialidase", "susceptibility"], "article_id"=>608510, "categories"=>["Virology"], "users"=>["Teruaki Nakatsuji", "Yu-Tsueng Liu", "Cheng-Po Huang", "Richard L. Gallo", "Chun-Ming Huang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0001551.g002", "stats"=>{"downloads"=>2, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Removal_of_Sialic_Acids_from_Human_Sebocytes_by_Sialidase_Increases_Their_Susceptibility_to_P_acnes_/608510", "title"=>"Removal of Sialic Acids from Human Sebocytes by Sialidase Increases Their Susceptibility to <i>P. acnes.</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-02-06 02:21:50"}
  • {"files"=>["https://ndownloader.figshare.com/files/938448"], "description"=>"<p>(A) For <i>in vitro</i> neutralization assays, <i>P. acnes</i> was pre-treated with 2.5% (v/v) anti-sialidase or GFP sera at 37°C for 2 h. Serum-pretreated <i>P. acnes</i> (2×10<sup>6</sup> CFU) was then co-cultured with sebocytes (2×10<sup>5</sup> cells) for 18 h. Sebocyte death induced by serum-pretreated <i>P. acnes</i> was detected by <i>p</i>NPP. The cytotoxicity of <i>P. acnes</i> was calculated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001551#s2\" target=\"_blank\">Methods</a> and presented as mean±SE (n = 4, <i>P</i><0.0005** by Student's <i>t</i>-test). (B) Serum-pretreated <i>P. acnes</i> (1.5×10<sup>8</sup> CFU) was co-cultured with sebocytes (3×10<sup>6</sup> cells) for 8 h. Measurement of IL-8 in the culture medium was carried out by ELISA assays using a Quantikine human IL-8 set (R&D System). The data is presented as mean±SE (n = 4, <i>P</i><0.01* by Student's <i>t</i>-test).</p>", "links"=>[], "tags"=>["anti-sialidase", "serum", "decreases", "il-8"], "article_id"=>608891, "categories"=>["Virology"], "users"=>["Teruaki Nakatsuji", "Yu-Tsueng Liu", "Cheng-Po Huang", "Richard L. Gallo", "Chun-Ming Huang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0001551.g006", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Neutralization_with_Anti_sialidase_Serum_Decreases_P_acnes_induced_Cell_Death_and_IL_8_Production_in_Human_Sebocytes_/608891", "title"=>"Neutralization with Anti-sialidase Serum Decreases <i>P. acnes</i>-induced Cell Death and IL-8 Production in Human Sebocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-02-06 02:28:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/937948"], "description"=>"<p>(A) A vector encoding a cell wall anchored sialidase (accession # gi|50843035) was constructed by inserting a PCR amplified full sialidase gene into the pEcoli-Nterm 6xHN vector (Clontech Laboratories, Inc., Mountain View, CA) at the SaII and HindIII restriction sites. Specific primers including the sense (5′- ATGACTTTGACCACGAAACTGAGCG-3′) and anti-sense primers (5′-TCAGGCAGGGCTCCGGCCCCAGATGC-3′) were designed to clone sialidase from <i>P. acnes</i> (ATCC 6919). The vector, which contains T7/LacO promoter, is derived from the pET system developed by William Studier and colleagues to achieve exceptionally high levels of protein expression in <i>E. coli</i>. Sialidase (arrow) was expressed in <i>E. coli</i> in the absence (lane 1) or presence (lane 2) of (1 mM) IPTG. After IPTG induction, sialidase was successfully expressed in <i>E. coli</i> and shown at about 53.1 kDa on a 10 % SDS-PAGE (arrowhead). (B) Purified sialidase (arrow) was obtained via In-Fusion Ready TALON Express Bacterial Expression and Purification kit (Clontech Laboratories, Inc., Mountain View, CA). (C) The expression and purity of sialidase were confirmed by Nano-LTQ MS/MS mass spectrometry (Thermo Electron Corp. Waltham, MA). A sequenced internal peptide (VVELSDTLMLNSR) of sialidase was presented.</p>", "links"=>[], "tags"=>["purification", "anchored"], "article_id"=>608393, "categories"=>["Virology"], "users"=>["Teruaki Nakatsuji", "Yu-Tsueng Liu", "Cheng-Po Huang", "Richard L. Gallo", "Chun-Ming Huang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0001551.g001", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_and_Purification_of_A_Cell_Wall_Anchored_Sialidase_/608393", "title"=>"Expression and Purification of A Cell Wall Anchored Sialidase.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-02-06 02:19:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/938378"], "description"=>"<p>(A) A tissue chamber (a) (internal and external diameters, 1.5 and 3 mm, respectively, length, 1 cm; internal volume, 80 µl) was implanted subcutaneously in the abdomen of sialidase- or GFP-immunized mice. The tissue chamber consisted of closed ploytetrafluoroethylene Teflon cylinders with 12 regularly spaced 0.1 mm holes. Bar: 1 cm. H&E staining of the cross-section of an implanted tissue chamber showed that the chamber was completely encapsulated by fibrotic tissue 7-days after implantation (b). Bar: 1.0 mm. <i>P. acnes</i> (10<sup>7</sup> CFU, 20 µl) was injected and trapped in this encapsulated tissue chamber. (B) Tissue chamber fluid containing MIP-2 was drawn by pecutaneous aspiration before (open bar) and three days after (solid bar) <i>P. acnes</i> injection. Measurement of MIP-2 was carried out by sandwich ELISA that used the Quantikine M mouse MIP-2 set (R&D System, Minneapolis, MN). Vaccination with sialidase markedly suppressed the <i>P. acnes</i>-induced increase in MIP-2. Error bars represented mean±SE of five separate experiments (*<i>P</i><0.005 by Student's <i>t</i>-test).</p>", "links"=>[], "tags"=>["sialidase-based", "acne", "suppresses", "induction", "pro-inflammatory", "cytokine", "mip-2"], "article_id"=>608821, "categories"=>["Virology"], "users"=>["Teruaki Nakatsuji", "Yu-Tsueng Liu", "Cheng-Po Huang", "Richard L. Gallo", "Chun-Ming Huang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0001551.g005", "stats"=>{"downloads"=>6, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_A_Sialidase_based_Acne_Vaccine_Suppresses_Induction_of_the_Pro_inflammatory_Cytokine_MIP_2_by_P_acnes_/608821", "title"=>"A Sialidase-based Acne Vaccine Suppresses Induction of the Pro-inflammatory Cytokine MIP-2 by <i>P. acnes.</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-02-06 02:27:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/462149", "https://ndownloader.figshare.com/files/462175", "https://ndownloader.figshare.com/files/462188"], "description"=>"<div><h3>Background</h3><p>Acne vulgaris afflicts more than fifty million people in the United State and the severity of this disorder is associated with the immune response to <em>Propionibacterium acnes (P. acnes)</em>. Systemic therapies for acne target <em>P. acnes</em> using antibiotics, or target the follicle with retinoids such as isotretinoin. The latter systemic treatment is highly effective but also carries a risk of side effects including immune imbalance, hyperlipidemia, and teratogenicity. Despite substantial research into potential new therapies for this common disease, vaccines against acne vulgaris are not yet available.</p><h3>Methods and Findings</h3><p>Here we create an acne vaccine targeting a cell wall-anchored sialidase of <em>P. acnes.</em> The importance of sialidase to disease pathogenesis is shown by treatment of a human sebocyte cell line with recombinant sialidase that increased susceptibility to <em>P. acnes</em> cytotoxicity and adhesion. Mice immunized with sialidase elicit a detectable antibody; the anti-sialidase serum effectively neutralized the cytotoxicity of <em>P. acnes in vitro</em> and <em>P. acnes</em>-induced interleukin-8 (IL-8) production in human sebocytes. Furthermore, the sialidase-immunized mice provided protective immunity against <em>P. acnes in vivo</em> as this treatment blocked an increase in ear thickness and release of pro-inflammatory macrophage inflammatory protein (MIP-2) cytokine.</p><h3>Conclusions</h3><p>Results indicated that acne vaccines open novel therapeutic avenues for acne vulgaris and other <em>P. acnes</em>-associated diseases.</p></div>", "links"=>[], "tags"=>["vaccination", "targeting", "sialidase", "acne", "vulgaris"], "article_id"=>151075, "categories"=>["Cancer"], "users"=>["Teruaki Nakatsuji", "Yu-Tsueng Liu", "Cheng-Po Huang", "Richard L. Gallo", "Chun-Ming Huang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0001551.s001", "https://dx.doi.org/10.1371/journal.pone.0001551.s002", "https://dx.doi.org/10.1371/journal.pone.0001551.s003"], "stats"=>{"downloads"=>3, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Vaccination_Targeting_a_Surface_Sialidase_of_P_acnes_Implication_for_New_Treatment_of_Acne_Vulgaris/151075", "title"=>"Vaccination Targeting a Surface Sialidase of <em>P. acnes:</em> Implication for New Treatment of Acne Vulgaris", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2008-02-06 00:17:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/938286"], "description"=>"<p>(A) The cytotoxicity of <i>P. acnes</i> was calculated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001551#s2\" target=\"_blank\">Methods</a> and presented as mean±SE (<i>P</i><0.0005** by Student's <i>t</i>-test). For assaying <i>in vivo</i> immune protection, ICR mice were immunized with recombinant sialidase or GFP using Freund (In)complete adjuvants (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0001551#pone-0001551-g003\" target=\"_blank\">Figure 3</a>). After confirming antibody production by western blot, live <i>P. acnes</i> (10<sup>7</sup> CFU, 25 µl) were injected subcutaneously into the ears of sialidase- and GFP-immunized mice, with PBS (25 µl) as a control. Ear thickness was periodically measured for 71 days after injection and changes reported as % of ear thickness in PBS-injected ears. <i>P. acnes</i>-induced ear swelling was significantly suppressed in sialidase-immunized mice in comparison with GFP-immunized mice (<i>P</i><0.05*), except for day 0, 4, 5 and 71. (B) Erythema was assessed in sialidase- (a) or GFP- (b) immunized mice 24 h after live <i>P. acnes</i> (left ears) or PBS (right ears) injection.</p>", "links"=>[], "tags"=>["conferred", "sialidase-based", "acne"], "article_id"=>608734, "categories"=>["Virology"], "users"=>["Teruaki Nakatsuji", "Yu-Tsueng Liu", "Cheng-Po Huang", "Richard L. Gallo", "Chun-Ming Huang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0001551.g004", "stats"=>{"downloads"=>0, "page_views"=>32, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Immune_Protection_Conferred_by_A_Sialidase_based_Acne_Vaccine_/608734", "title"=>"Immune Protection Conferred by A Sialidase-based Acne Vaccine.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-02-06 02:25:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/938166"], "description"=>"<p>(A) ICR mice were vaccinated with heat-killed <i>P. acnes</i> for nine weeks (three boosts at a three-week interval). Serum (1: 500 dilution), harvested one week after the third boost, was reacted to recombinant sialidase (1 µg; lane 1), GFP (1 µg, lane 2), and <i>P. acnes</i> lysates (7 µg, lane 3) that had been run on a 10% SDS-PAGE. The 6x HN tag of recombinant sialidase was removed by enterokinase before loading into a SDS-PAGE. Although heat-killed <i>P. acnes</i>-vaccinated mice produced anti-sera against several <i>P. acnes</i> proteins (>50 kDa) (lane 3), sialidase specific antibodies were not detected (lane 3). (B) ICR mice were then vaccinated with a recombinant sialidase (lane 1) or GFP (lane 2) using Freund/(in)complete adjuvants. For the first vaccination, mice were subcutaneously inoculated with 50 µg of sialidase or GFP, which were emulsified with a complete Freund adjuvant. Two weeks later, the second vaccination was delivered via intramuscular injection with the same amount of antigen mixed well with incomplete Freund adjuvant. Anti-sialidase antibodies were detected by western blot one week after the second vaccination. 1 µg of recombinant sialidase or GFP were separated via 10 % SDS-PAGE, transferred to a PVDF membrane and reacted with mouse sera (10,000-fold dilution). Sialidase antibodies were provoked when mice were immunized with recombinant sialidase, but not with GFP. Data is representative of three separate experiments with similar results.</p>", "links"=>[], "tags"=>["immunized", "recombinant", "sialidase", "freund", "adjuvants"], "article_id"=>608605, "categories"=>["Virology"], "users"=>["Teruaki Nakatsuji", "Yu-Tsueng Liu", "Cheng-Po Huang", "Richard L. Gallo", "Chun-Ming Huang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0001551.g003", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Mice_Immunized_with_Recombinant_Sialidase_and_Freund_in_complete_Adjuvants_Produce_Sialidase_Specific_Antibodies_/608605", "title"=>"Mice Immunized with Recombinant Sialidase and Freund (in)complete Adjuvants Produce Sialidase Specific Antibodies.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2008-02-06 02:23:25"}

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Relative Metric

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