CMV-Independent Lysis of Glioblastoma by Ex Vivo Expanded/Activated Vδ1+ γδ T Cells
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{"title"=>"CMV-Independent Lysis of Glioblastoma by Ex Vivo Expanded/Activated Vδ1+ γδ T Cells", "type"=>"journal", "authors"=>[{"first_name"=>"Andrea", "last_name"=>"Knight", "scopus_author_id"=>"8414779300"}, {"first_name"=>"Hilal", "last_name"=>"Arnouk", "scopus_author_id"=>"9335986800"}, {"first_name"=>"William", "last_name"=>"Britt", "scopus_author_id"=>"35449543100"}, {"first_name"=>"G. Yancey", "last_name"=>"Gillespie", "scopus_author_id"=>"7102523969"}, {"first_name"=>"Gretchen A.", "last_name"=>"Cloud", "scopus_author_id"=>"24316041100"}, {"first_name"=>"Lualhati", "last_name"=>"Harkins", "scopus_author_id"=>"56798022300"}, {"first_name"=>"Yun", "last_name"=>"Su", "scopus_author_id"=>"55554178200"}, {"first_name"=>"Mark W.", "last_name"=>"Lowdell", "scopus_author_id"=>"7003930276"}, {"first_name"=>"Lawrence S.", "last_name"=>"Lamb", "scopus_author_id"=>"7005353427"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84881326207", "doi"=>"10.1371/journal.pone.0068729", "issn"=>"19326203", "pui"=>"369535360", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"23950874", "scopus"=>"2-s2.0-84881326207"}, "id"=>"bbc3ab36-f1d9-3161-94aa-246719a4c165", "abstract"=>"Vδ2(neg) γδ T cells, of which Vδ1+ γδ T cells are by far the largest subset, are important effectors against CMV infection. Malignant gliomas often contain CMV genetic material and proteins, and evidence exists that CMV infection may be associated with initiation and/or progression of glioblastoma multiforme (GBM). We sought to determine if Vδ1+ γδ T cells were cytotoxic to GBM and the extent to which their cytotoxicity was CMV dependent. We examined the cytotoxic effect of ex vivo expanded/activated Vδ1+ γδ T cells from healthy CMV seropositive and CMV seronegative donors on unmanipulated and CMV-infected established GBM cell lines and cell lines developed from short- term culture of primary tumors. Expanded/activated Vδ1+ T cells killed CMV-negative U251, U87, and U373 GBM cell lines and two primary tumor explants regardless of the serologic status of the donor. Experimental CMV infection did not increase Vδ1+ T cell--mediated cytotoxicity and in some cases the cell lines were more resistant to lysis when infected with CMV. Flow cytometry analysis of CMV-infected cell lines revealed down-regulation of the NKG2D ligands ULBP-2, and ULBP-3 as well as MICA/B in CMV-infected cells. These studies show that ex vivo expanded/activated Vδ1+ γδ T cells readily recognize and kill established GBM cell lines and primary tumor-derived GBM cells regardless of whether CMV infection is present, however, CMV may enhance the resistance GBM cell lines to innate recognition possibly contributing to the poor immunogenicity of GBM.", "link"=>"http://www.mendeley.com/research/cmvindependent-lysis-glioblastoma-ex-vivo-expandedactivated-v%CE%B41-%CE%B3%CE%B4-t-cells", "reader_count"=>23, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Student > Master"=>4, "Student > Bachelor"=>5, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>7, "Student > Postgraduate"=>1, "Student > Master"=>4, "Student > Bachelor"=>5, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>11, "Medicine and Dentistry"=>6, "Immunology and Microbiology"=>3}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>6}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>11}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"United States"=>3, "Russia"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1146361"], "description"=>"<p>(a) Primary GBM from which cell explants 1016 and 1042 were derived expressed CMV-associated surface antigens IE-1 and pp65, although T cell invasion beyond perivascular areas as indicated by anti-CD3 labeling is minimal. (b) Both primary tumors 1016 and 1042 (top panel) were assessed for the presence of CMV mRNA by in-situ hybridization using a biotinylated 21-base oligonucleotide (5′-GTGGTGGCGCTGGGGGTGGCG-3′) specific for HCMV early gene mRNA and a biotinylated positive (specific for polyadenylic mRNA) and negative control (specific for HSV-1/2) probe. provided positive (specific for endogenous alu DNA sequence) and negative (nonspecific DNA) digoxigenin-labeled control probes. Both tumors show the presence of CMV mRNA as do the short passage cell lines that were derived from them (bottom panel) suggesting continued CMV infection.</p>", "links"=>[], "tags"=>["immunology", "immunity", "immunotherapy", "Clinical immunology", "Immune cells", "t cells", "Immunologic subspecialties", "Tumor immunology", "Immune response", "Infectious diseases", "Viral diseases", "Cytomegalovirus infection", "oncology", "Cancer treatment", "Bone marrow transplantation", "Cancers and neoplasms", "Neurological tumors", "Glioblastoma multiforme", "cmv", "gbm", "derived"], "article_id"=>767067, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Andrea Knight", "Hilal Arnouk", "William Britt", "G. Yancey Gillespie", "Gretchen A. Cloud", "Lualhati Harkins", "Yun Su", "Mark W. Lowdell", "Lawrence S. Lamb"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068729.g004", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Assessment_of_CMV_activity_in_primary_GBM_and_derived_cell_lines_/767067", "title"=>"Assessment of CMV activity in primary GBM and derived cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-07 02:59:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1146363"], "description"=>"<p>(a) Lysis of unmanipulated U251MG cells (filled circles) versus CMV-infected U251MG cells (open circles) by expanded/activated Vδ1+ T cells for three representative donors in a 4 h flow cytometric cytotoxicity assay. CMV-infected cells are more resistant to lysis (p = 0.021) at E∶T ratio of 20∶1. (b) Expression of NKG2DL on three cultures of unmanipulated (CMV-) vs. CMV-infected (CMV+) U251 glioma cultures as determined by Median Fluorescence Intensity (MFI) following acquisition of a minimum of 10,000 events. (c) Expression of NKG2DL on unmanipulated (black), HCMV IE-1 expressing (red) and IE-1<sup>neg</sup> (blue) U251 glioma cells after 5 days of culture (see text).</p>", "links"=>[], "tags"=>["immunology", "immunity", "immunotherapy", "Clinical immunology", "Immune cells", "t cells", "Immunologic subspecialties", "Tumor immunology", "Immune response", "Infectious diseases", "Viral diseases", "Cytomegalovirus infection", "oncology", "Cancer treatment", "Bone marrow transplantation", "Cancers and neoplasms", "Neurological tumors", "Glioblastoma multiforme", "modulated", "nkg2dl", "glioma", "lines", "vulnerability", "lysis"], "article_id"=>767069, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Andrea Knight", "Hilal Arnouk", "William Britt", "G. Yancey Gillespie", "Gretchen A. Cloud", "Lualhati Harkins", "Yun Su", "Mark W. Lowdell", "Lawrence S. Lamb"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068729.g006", "stats"=>{"downloads"=>1, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CMV_infection_modulated_NKG2DL_expression_on_glioma_cell_lines_and_their_vulnerability_to_lysis_by_V_948_1_T_cells_/767069", "title"=>"CMV infection modulated NKG2DL expression on glioma cell lines and their vulnerability to lysis by Vδ1+ T cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-07 02:59:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1146362"], "description"=>"<p>Primary GBM cell line explants 1016 and 1042 were labeled with fluorochrome-tagged antibodies to respective isotype control anti-IgG (clear) and Vδ1+ T cell target NKG2DL MIC A/B, ULBP-1, ULBP-2 and ULBP-3 (shaded). Strong expression of ULBP-2 and ULBP-3 is noted. Primary 1042 is a heterogeneous tumor with a brightly autofluorescent subpopulation seen as a bimodal peak for both isotype and labeled plots.</p>", "links"=>[], "tags"=>["immunology", "immunity", "immunotherapy", "Clinical immunology", "Immune cells", "t cells", "Immunologic subspecialties", "Tumor immunology", "Immune response", "Infectious diseases", "Viral diseases", "Cytomegalovirus infection", "oncology", "Cancer treatment", "Bone marrow transplantation", "Cancers and neoplasms", "Neurological tumors", "Glioblastoma multiforme", "antigen", "lines", "derived"], "article_id"=>767068, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Andrea Knight", "Hilal Arnouk", "William Britt", "G. Yancey Gillespie", "Gretchen A. Cloud", "Lualhati Harkins", "Yun Su", "Mark W. Lowdell", "Lawrence S. Lamb"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068729.g005", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Assessment_of_stress_antigen_expression_cell_lines_derived_from_primary_GBM_/767068", "title"=>"Assessment of stress antigen expression cell lines derived from primary GBM.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-07 02:59:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1146359"], "description"=>"<p>(<b><i>a</i></b>) Flow cytometric analysis of 14-day expansion culture of purified Vδ1+ T cells from two healthy volunteer donors. (<b><i>b</i></b>) Cytotoxicity of purified and expanded Vδ1+ T cells from two donors against three (left to right for each donor) GBM cell lines (<i>a</i>) U251, (<i>b</i>) U87, and (<i>c</i>) U373 and two primary cell lines from GBM explants designated (d)1016 and (<i>e</i>)1042 in a four-hour cytotoxicity assay at an E∶T ratio of 20∶1. (<b><i>c</i></b>) Vδ1+ T cell cultures were expanded from additional CMV-seropositive (n = 4) and CMV-seronegative (n = 6) donors as described in the text and reacted over 4 h at an E∶T ratio of 20∶1 is shown with the U251MG glioma cell line and the 1024 primary GBM explant line. Donor CMV serologic status had no effect on Vδ1+ T cell cytotoxicity against U251 (p = 0.75) or 1042 (p = 0.29), respectively for CMV seropositive and CMV seronegative donors.</p>", "links"=>[], "tags"=>["immunology", "immunity", "immunotherapy", "Clinical immunology", "Immune cells", "t cells", "Immunologic subspecialties", "Tumor immunology", "Immune response", "Infectious diseases", "Viral diseases", "Cytomegalovirus infection", "oncology", "Cancer treatment", "Bone marrow transplantation", "Cancers and neoplasms", "Neurological tumors", "Glioblastoma multiforme", "cells", "cmv", "seropositive", "seronegative", "volunteers", "glioma"], "article_id"=>767065, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Andrea Knight", "Hilal Arnouk", "William Britt", "G. Yancey Gillespie", "Gretchen A. Cloud", "Lualhati Harkins", "Yun Su", "Mark W. Lowdell", "Lawrence S. Lamb"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068729.g003", "stats"=>{"downloads"=>2, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cytotoxicity_of_expanded_activated_V_948_1_T_cells_from_CMV_seropositive_and_seronegative_healthy_volunteers_against_human_glioma_cell_lines_/767065", "title"=>"Cytotoxicity of expanded/activated Vδ1+ T cells from CMV seropositive and seronegative healthy volunteers against human glioma cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-07 02:59:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1146357"], "description"=>"<p>The γδ T cells were expanded/activated by either <i>(a)</i> CD2/OKT-3 or <i>(b)</i> ZOL/IL-2 methods as described in the text. Peripheral blood from a representative healthy control is shown at right. After 14 days in culture, neither method was effective in expanding γδ T cells from the blood of any of the five GBM patients although γδ T cells from healthy volunteers were readily induced to proliferate (p = 0.12 for both methods).</p>", "links"=>[], "tags"=>["immunology", "immunity", "immunotherapy", "Clinical immunology", "Immune cells", "t cells", "Immunologic subspecialties", "Tumor immunology", "Immune response", "Infectious diseases", "Viral diseases", "Cytomegalovirus infection", "oncology", "Cancer treatment", "Bone marrow transplantation", "Cancers and neoplasms", "Neurological tumors", "Glioblastoma multiforme", "cytometry", "cultures", "peripheral", "pre-resection", "gbm", "patients"], "article_id"=>767063, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Andrea Knight", "Hilal Arnouk", "William Britt", "G. Yancey Gillespie", "Gretchen A. Cloud", "Lualhati Harkins", "Yun Su", "Mark W. Lowdell", "Lawrence S. Lamb"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068729.g001", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Flow_cytometry_dot_plots_947_948_T_cell_expansion_cultures_of_peripheral_blood_from_five_pre_resection_GBM_patients_and_one_healthy_volunteer_/767063", "title"=>"Flow cytometry dot plots γδ T cell expansion cultures of peripheral blood from five pre-resection GBM patients and one healthy volunteer.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-07 02:59:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1146358"], "description"=>"<p>Note an initial very low percentage of γδ T cells that could not be expanded in two week culture using the CD2/OKT3 method (see text). Flow cytometric dot plots were gated on CD3+ lymphocytes. Similar results were obtained from three additional patients.</p>", "links"=>[], "tags"=>["immunology", "immunity", "immunotherapy", "Clinical immunology", "Immune cells", "t cells", "Immunologic subspecialties", "Tumor immunology", "Immune response", "Infectious diseases", "Viral diseases", "Cytomegalovirus infection", "oncology", "Cancer treatment", "Bone marrow transplantation", "Cancers and neoplasms", "Neurological tumors", "Glioblastoma multiforme", "infiltrating", "lymphocytes"], "article_id"=>767064, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Andrea Knight", "Hilal Arnouk", "William Britt", "G. Yancey Gillespie", "Gretchen A. Cloud", "Lualhati Harkins", "Yun Su", "Mark W. Lowdell", "Lawrence S. Lamb"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068729.g002", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Culture_of_tumor_infiltrating_lymphocytes_TIL_from_a_representative_human_GBM_/767064", "title"=>"Culture of tumor infiltrating lymphocytes (TIL) from a representative human GBM.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-07 02:59:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/1146365"], "description"=>"<div><p>Vδ2<sup>neg</sup> γδ T cells, of which Vδ1+ γδ T cells are by far the largest subset, are important effectors against CMV infection. Malignant gliomas often contain CMV genetic material and proteins, and evidence exists that CMV infection may be associated with initiation and/or progression of glioblastoma multiforme (GBM). We sought to determine if Vδ1+ γδ T cells were cytotoxic to GBM and the extent to which their cytotoxicity was CMV dependent. We examined the cytotoxic effect of <i>ex vivo</i> expanded/activated Vδ1+ γδ T cells from healthy CMV seropositive and CMV seronegative donors on unmanipulated and CMV-infected established GBM cell lines and cell lines developed from short- term culture of primary tumors. Expanded/activated Vδ1+ T cells killed CMV-negative U251, U87, and U373 GBM cell lines and two primary tumor explants regardless of the serologic status of the donor. Experimental CMV infection did not increase Vδ1+ T cell - mediated cytotoxicity and in some cases the cell lines were more resistant to lysis when infected with CMV. Flow cytometry analysis of CMV-infected cell lines revealed down-regulation of the NKG2D ligands ULBP-2, and ULBP-3 as well as MICA/B in CMV-infected cells. These studies show that <i>ex vivo</i> expanded/activated Vδ1+ γδ T cells readily recognize and kill established GBM cell lines and primary tumor-derived GBM cells regardless of whether CMV infection is present, however, CMV may enhance the resistance GBM cell lines to innate recognition possibly contributing to the poor immunogenicity of GBM.</p></div>", "links"=>[], "tags"=>["immunology", "immunity", "immunotherapy", "Clinical immunology", "Immune cells", "t cells", "Immunologic subspecialties", "Tumor immunology", "Immune response", "Infectious diseases", "Viral diseases", "Cytomegalovirus infection", "oncology", "Cancer treatment", "Bone marrow transplantation", "Cancers and neoplasms", "Neurological tumors", "Glioblastoma multiforme", "cmv-independent", "lysis", "glioblastoma", "cells"], "article_id"=>767071, "categories"=>["Medicine", "Biological Sciences"], "users"=>["Andrea Knight", "Hilal Arnouk", "William Britt", "G. Yancey Gillespie", "Gretchen A. Cloud", "Lualhati Harkins", "Yun Su", "Mark W. Lowdell", "Lawrence S. Lamb"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0068729", "stats"=>{"downloads"=>5, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/CMV_Independent_Lysis_of_Glioblastoma_by_Ex_Vivo_Expanded_Activated_V_1_T_Cells/767071", "title"=>"CMV-Independent Lysis of Glioblastoma by <i>Ex Vivo</i> Expanded/Activated Vδ1+ γδ T Cells", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-08-07 02:59:16"}

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Relative Metric

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