Role of Myeloid-Derived Suppressor Cells in Amelioration of Experimental Autoimmune Hepatitis Following Activation of TRPV1 Receptors by Cannabidiol
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{"title"=>"Role of Myeloid-derived suppressor cells in amelioration of experimental autoimmune hepatitis following activation of TRPV1 receptors by cannabidiol", "type"=>"journal", "authors"=>[{"first_name"=>"Venkatesh L.", "last_name"=>"Hegde", "scopus_author_id"=>"8634017500"}, {"first_name"=>"Prakash S.", "last_name"=>"Nagarkatti", "scopus_author_id"=>"7005792287"}, {"first_name"=>"Mitzi", "last_name"=>"Nagarkatti", "scopus_author_id"=>"7005891809"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-79953658580", "pui"=>"361550367", "doi"=>"10.1371/journal.pone.0018281", "isbn"=>"1932-6203", "sgr"=>"79953658580", "pmid"=>"21483776"}, "id"=>"42ec2b39-82a0-393b-a215-01fb83a1d071", "abstract"=>"BACKGROUND: Myeloid-derived suppressor cells (MDSCs) are getting increased attention as one of the main regulatory cells of the immune system. They are induced at sites of inflammation and can potently suppress T cell functions. In the current study, we demonstrate how activation of TRPV1 vanilloid receptors can trigger MDSCs, which in turn, can inhibit inflammation and hepatitis.\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: Polyclonal activation of T cells, following injection of concanavalin A (ConA), in C57BL/6 mice caused acute hepatitis, characterized by significant increase in aspartate transaminase (AST), induction of inflammatory cytokines, and infiltration of mononuclear cells in the liver, leading to severe liver injury. Administration of cannabidiol (CBD), a natural non-psychoactive cannabinoid, after ConA challenge, inhibited hepatitis in a dose-dependent manner, along with all of the associated inflammation markers. Phenotypic analysis of liver infiltrating cells showed that CBD-mediated suppression of hepatitis was associated with increased induction of arginase-expressing CD11b(+)Gr-1(+) MDSCs. Purified CBD-induced MDSCs could effectively suppress T cell proliferation in vitro in arginase-dependent manner. Furthermore, adoptive transfer of purified MDSCs into naïve mice conferred significant protection from ConA-induced hepatitis. CBD failed to induce MDSCs and suppress hepatitis in the livers of vanilloid receptor-deficient mice (TRPV1(-/-)) thereby suggesting that CBD primarily acted via this receptor to induce MDSCs and suppress hepatitis. While MDSCs induced by CBD in liver consisted of granulocytic and monocytic subsets at a ratio of ∼2∶1, the monocytic MDSCs were more immunosuppressive compared to granulocytic MDSCs. The ability of CBD to induce MDSCs and suppress hepatitis was also demonstrable in Staphylococcal enterotoxin B-induced liver injury.\\n\\nCONCLUSIONS/SIGNIFICANCE: This study demonstrates for the first time that MDSCs play a critical role in attenuating acute inflammation in the liver, and that agents such as CBD, which trigger MDSCs through activation of TRPV1 vanilloid receptors may constitute a novel therapeutic modality to treat inflammatory diseases.", "link"=>"http://www.mendeley.com/research/role-myeloidderived-suppressor-cells-amelioration-experimental-autoimmune-hepatitis-following-activa", "reader_count"=>38, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>7, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>4, "Student > Bachelor"=>4, "Professor"=>4}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Librarian"=>1, "Researcher"=>7, "Student > Ph. D. Student"=>10, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>4, "Student > Bachelor"=>4, "Professor"=>4}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>13, "Medicine and Dentistry"=>6, "Neuroscience"=>1, "Psychology"=>3, "Immunology and Microbiology"=>6}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>6}, "Neuroscience"=>{"Neuroscience"=>1}, "Psychology"=>{"Psychology"=>3}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>6}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>13}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}, "Unspecified"=>{"Unspecified"=>4}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "United States"=>1, "United Kingdom"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/786077"], "description"=>"<p>WT mice (n = 6) were sensitized by injecting\n D-galactosamine (GalN), 20 mg/mouse in 100 µL PBS\n (<i>i.p.</i>). After 15 min, SEB was injected (40\n µg/mouse, in 100 µL PBS, <i>i.p.</i>). For\n treatment groups, 10 or 50 mg/kg bd.wt. of CBD was administered\n <i>i.p.</i> A) Blood was collected at 8 h post SEB\n injection and serum AST levels were measured. Student's\n <i>t</i>-test (<sup>#</sup>p<0.01 compared to Veh/GalN\n control; *p<0.05, **p<0.01 compared to GalN/SEB). B.\n <b>Histology:</b> Liver samples after 48 h were fixed and\n embedded in paraffin. Five µm sections were stained by H&E and\n analyzed by light microscopy. Representative photomicrographs are shown.\n a) Veh/GalN, b) CBD50, c) GalN/SEB and d) GalN/SEB+CBD50 (CBD50\n  = 50 mg/kg). Liver infiltrating cells from the\n treatment groups as indicated were isolated at 24 h and stained for\n CD11b and Gr-1 and analyzed by flow cytometry\n (CBD25 = 25 mg/kg). Representative dot plots are\n shown with percentages of CD11b<sup>+</sup>Gr-1<sup>+</sup>\n MDSCs (C). D) Absolute number of MDSCs in liver for each group\n (n = 4). Student's <i>t</i>-test\n (**p<0.01 compared to GalN/SEB).</p>", "links"=>[], "tags"=>["attenuates", "seb-induced", "acute", "inducing", "mdscs", "in"], "article_id"=>456449, "categories"=>["Immunology"], "users"=>["Venkatesh L. Hegde", "Prakash S. Nagarkatti", "Mitzi Nagarkatti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0018281.g008", "stats"=>{"downloads"=>2, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CBD_attenuates_SEB_induced_acute_liver_injury_by_inducing_MDSCs_in____liver_/456449", "title"=>"CBD attenuates SEB-induced acute liver injury by inducing MDSCs in\n liver.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 19:25:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/785475"], "description"=>"<p>WT mice (n = 6) were injected with PBS (control), or\n ConA (12.5 mg/kg bd.wt., <i>i.v.</i>) to induce hepatitis.\n ConA injected mice were administered <i>i.p.</i> with vehicle\n or CBD at 5, 20 or 50 mg/kg bd.wt. CBD group received CBD alone at 50\n mg/kg. A) Blood was collected at 6, 12, 24 and 48 h and serum aspartate\n transaminase (AST) was determined by spectrophotometric assay. Data\n represent mean ± SEM from 6 individual mice. The changes in AST\n levels were analyzed by Student's <i>t</i>-test (#\n p<0.01 compared to PBS; *p<0.05, **p<0.01 compared\n to ConA+veh). <b>Histological analysis:</b> B. Representative\n H&E stained liver sections obtained 24 h post ConA injection in WT\n mice (original magnification ×100). a) Vehicle and b) CBD alone,\n show normal tissue morphology; c) ConA+veh shows large necrotic\n lesion induced by ConA (arrows); d) ConA+CBD (50 mg/kg bd.wt.)\n shows less necrosis (arrows) of liver parenchyma. C) Quantification of\n necrotic lesions as a percentage of liver parenchyma. Data are mean\n ± SEM from 4 mice per treatment. At least five fields were\n analyzed per section from each liver sample. PBS and CBD controls did\n not show any necrotic lesions. Student's <i>t</i>-test,\n **p<0.01 compared to ConA.</p>", "links"=>[], "tags"=>["attenuates", "cona-induced", "hepatitis"], "article_id"=>455848, "categories"=>["Immunology"], "users"=>["Venkatesh L. Hegde", "Prakash S. Nagarkatti", "Mitzi Nagarkatti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0018281.g001", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cannabidiol_CBD_attenuates_ConA_induced_hepatitis_in_mice_/455848", "title"=>"Cannabidiol (CBD) attenuates ConA-induced hepatitis in mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 19:22:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/785569"], "description"=>"<p>A) CBD acts by suppressing pro-inflammatory cytokines in ConA-induced\n hepatitis. WT mice (n = 6) were injected with PBS\n or ConA. ConA injected mice were administered with vehicle or CBD (25\n mg/kg bd.wt.). CBD group received CBD alone at 25 mg/kg bd.wt. Blood was\n collected after 12 h and serum cytokines were determined by Bioplex\n assay. Error bars represent mean ± SEM\n (n = 6). The data were analyzed by Student's\n <i>t</i>-test (<sup>##</sup>p<0.01 compared to PBS;\n *p<0.05, **p<0.01 compared to ConA+Veh). B)\n Semi-quantitative RT-PCR for SOCS-1 and SOCS-3 in livers 2 h after\n various treatments <i>in vivo</i>. 18S was used as a loading\n control. C. The densities of bands were quantified using gel imaging\n system (BioRad) and expressed as percentage expression relative to 18S.\n Student's <i>t</i>-test, *p<0.05 compared to\n ConA.</p>", "links"=>[], "tags"=>["inflammatory", "socs1"], "article_id"=>455948, "categories"=>["Immunology"], "users"=>["Venkatesh L. Hegde", "Prakash S. Nagarkatti", "Mitzi Nagarkatti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0018281.g002", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_inflammatory_cytokines_SOCS1_and_SOCS3_/455948", "title"=>"Analysis of inflammatory cytokines, SOCS1 and SOCS3.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 19:22:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/785832"], "description"=>"<p>CD11b<sup>+</sup>Gr-1<sup>+</sup> cells from liver infiltrating\n cells of ConA+CBD injected mice were purified, irradiated and\n co-cultured at different ratios with naïve syngenic purified T\n cells in the presence of mitogen ConA (4 µg/mL) for 48 h. T cell\n proliferation was determined by [3H]thymidine incorporation\n during the last 8 h of culture. In some wells with T cell: MDSC ratio of\n 10∶1, arginase-1 inhibitor (nor-NOHA) was added at the start of\n the culture at a concentration of 100 µM. Data are mean ±\n SEM of triplicate determinations and representative of two independent\n experiments. (B) WT mice (n = 4) were injected with\n vehicle or ConA. Five million purified\n CD11b<sup>+</sup>Gr-1<sup>+</sup> cells isolated from\n livers of ConA+CBD group, were adoptively transferred 12 h before\n injecting ConA. Blood was collected 12 h post-ConA injection and\n analyzed for AST. Data represent mean ± SEM from 4 mice per\n treatment. Student's <i>t-</i>test (*p<0.05,\n **p<0.01).</p>", "links"=>[], "tags"=>["mdscs", "immunosuppressive"], "article_id"=>456204, "categories"=>["Immunology"], "users"=>["Venkatesh L. Hegde", "Prakash S. Nagarkatti", "Mitzi Nagarkatti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0018281.g005", "stats"=>{"downloads"=>2, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CBD_induced_MDSCs_in_liver_are_immunosuppressive_in_____vitro_and_in_vivo_/456204", "title"=>"CBD-induced MDSCs in liver are immunosuppressive <i>in\n vitro</i> and <i>in vivo.</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 19:24:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/785899"], "description"=>"<p>WT or TRPV1 vanilloid receptor knockout (VR1-KO) mice were injected with\n vehicle, CBD, ConA+veh or ConA+CBD. A) After 16 h, blood was\n collected and serum AST levels were measured. B) Liver infiltrating\n leukocytes were stained and analyzed for\n CD11b<sup>+</sup>Gr-1<sup>+</sup> MDSCs by FACS.\n Representative dot plots with percentage of MDSCs shown (gated). C)\n Absolute number of MDSCs in the liver of WT and VR1-KO mice. Data are\n mean ± SEM with 4 mice per treatment. **p<0.01,\n ***p<0.001, n.s. (not significant) based on\n Student's <i>t</i>-test.</p>", "links"=>[], "tags"=>["trpv1"], "article_id"=>456273, "categories"=>["Immunology"], "users"=>["Venkatesh L. Hegde", "Prakash S. Nagarkatti", "Mitzi Nagarkatti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0018281.g006", "stats"=>{"downloads"=>1, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Role_of_TRPV1_vanilloid_receptors_/456273", "title"=>"Role of TRPV1 (vanilloid) receptors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 19:24:48"}
  • {"files"=>["https://ndownloader.figshare.com/files/785657"], "description"=>"<p>WT mice were injected with PBS (Veh) or ConA. ConA injected mice were\n simultaneously administered (<i>i.p.</i>) with vehicle or CBD\n (25 mg/kg bd.wt.). CBD group received CBD alone at 25 mg/kg bd.wt. Liver\n infiltrating cells isolated after 12 h were stained for CD11b and Gr-1,\n and analyzed by FACS. Representative flow profiles for each sample\n showing frequency of CD11b<sup>+</sup>Gr-1<sup>+</sup> cells\n (gated) is shown in panel A. Absolute numbers of\n CD11b<sup>+</sup>Gr-1<sup>+</sup> cells were calculated\n from percentages and total cell numbers per liver (B). Cells were also\n stained and analyzed for Foxp3<sup>+</sup> Tregs (D, E). Data\n represent mean ± SEM from 4 mice per treatment. Data were\n analyzed by Student's <i>t</i>-test (*p<0.05,\n **p<0.01).</p>", "links"=>[], "tags"=>["characterization", "infiltrating", "cbd", "hepatitis", "increases", "predominantly", "numbers", "in"], "article_id"=>456021, "categories"=>["Immunology"], "users"=>["Venkatesh L. Hegde", "Prakash S. Nagarkatti", "Mitzi Nagarkatti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0018281.g003", "stats"=>{"downloads"=>4, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phenotypic_characterization_of_liver_infiltrating_cells_CBD____treatment_of_hepatitis_increases_predominantly_____CD11b_Gr_1_cell_numbers_in____liver_/456021", "title"=>"Phenotypic characterization of liver infiltrating cells: CBD\n treatment of hepatitis increases predominantly\n CD11b<sup>+</sup>Gr-1<sup>+</sup> cell numbers in\n liver.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 19:23:23"}
  • {"files"=>["https://ndownloader.figshare.com/files/785737"], "description"=>"<p>Liver infiltrating cells isolated from indicated treatment groups\n (n = 4) were triple-stained for CD11b, Gr-1 and\n intracellular arginase 1. Respective histograms (filled) for arginase\n expression on CD11b<sup>+</sup>Gr-1<sup>+</sup> gated cells\n are shown for each group with respective MFI (A). Open histograms\n represent staining control. B) Arginase functional activity was\n determined by spectrophotometric assay using lysates of liver\n infiltrating cells from each group. Data are mean ± SEM\n (n = 4). Student's <i>t</i>-test,\n **p<0.01. C) Immunohistochemistry for arginase expression in\n liver sections and D) Morphology of infiltrating liver cells by Wright\n Giemsa staining (a. Veh, b. CBD, c. ConA+veh, d.\n ConA+CBD).</p>", "links"=>[], "tags"=>["mdscs"], "article_id"=>456118, "categories"=>["Immunology"], "users"=>["Venkatesh L. Hegde", "Prakash S. Nagarkatti", "Mitzi Nagarkatti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0018281.g004", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_MDSCs_in_liver_/456118", "title"=>"Characterization of MDSCs in liver.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 19:23:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/785980"], "description"=>"<p>Liver infiltrating cells were isolated from ConA+CBD injected mice\n at 12 h. Cells were triple-stained for CD11b, Ly6-G and Ly-6C, and\n analyzed by FACS. A) Cells were gated for CD11b<sup>+</sup>\n expression (histogram) and further analyzed for Ly6-G and Ly6-C antigens\n (dot plot). Representative dot plot shows percentages of\n CD11b<sup>+</sup>Ly6-G<sup>+</sup>Ly6-C<sup>+</sup>\n granulocytic MDSCs (Gr-MDSC) and\n CD11b<sup>+</sup>Ly6-G<sup>–</sup>Ly6-C<sup>+</sup>\n monocytic MDSCs (Mo-MDSC). B & C) MDSC subsets were purified by FACS\n sorting and used in T cell proliferation assay <i>in vitro</i>\n at different ratios with syngenic purified T cells stimulated with ConA.\n T cell proliferation was assessed by [3H]thymidine\n incorporation at 48 h. T cells cultured without MDSCs and with ConA\n served as the positive control. Data are mean ± SEM for\n triplicate determinations and representative of two separate\n experiments. **p<0.01, ***p<0.001, based on\n Student's t test.</p>", "links"=>[], "tags"=>["mdsc", "subsets", "induced", "cbd", "their", "suppressive"], "article_id"=>456349, "categories"=>["Immunology"], "users"=>["Venkatesh L. Hegde", "Prakash S. Nagarkatti", "Mitzi Nagarkatti"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0018281.g007", "stats"=>{"downloads"=>2, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Identification_of_MDSC_subsets_induced_by_CBD_in_liver_and_their____suppressive_activity_/456349", "title"=>"Identification of MDSC subsets induced by CBD in liver and their\n suppressive activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 19:25:12"}

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