Engineering Antigen-Specific T Cells from Genetically Modified Human Hematopoietic Stem Cells in Immunodeficient Mice
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{"title"=>"Engineering antigen-specific T cells from genetically modified human hematopoietic stem cells in immunodeficient mice", "type"=>"journal", "authors"=>[{"first_name"=>"Scott G.", "last_name"=>"Kitchen", "scopus_author_id"=>"26643624800"}, {"first_name"=>"Michael", "last_name"=>"Bennett", "scopus_author_id"=>"57198410981"}, {"first_name"=>"Zoran", "last_name"=>"Galić", "scopus_author_id"=>"6506821706"}, {"first_name"=>"Joanne", "last_name"=>"Kim", "scopus_author_id"=>"35748723800"}, {"first_name"=>"Qing", "last_name"=>"Xu", "scopus_author_id"=>"55482780600"}, {"first_name"=>"Alan", "last_name"=>"Young", "scopus_author_id"=>"35749231200"}, {"first_name"=>"Alexis", "last_name"=>"Lieberman", "scopus_author_id"=>"35748992800"}, {"first_name"=>"Aviva", "last_name"=>"Joseph", "scopus_author_id"=>"7202236551"}, {"first_name"=>"Harris", "last_name"=>"Goldstein", "scopus_author_id"=>"7203024663"}, {"first_name"=>"Hwee", "last_name"=>"Ng", "scopus_author_id"=>"7401618653"}, {"first_name"=>"Otto", "last_name"=>"Yang", "scopus_author_id"=>"7006753492"}, {"first_name"=>"Jerome A.", "last_name"=>"Zack", "scopus_author_id"=>"7101844958"}], "year"=>2009, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"359087377", "issn"=>"19326203", "isbn"=>"1932-6203", "doi"=>"10.1371/journal.pone.0008208", "scopus"=>"2-s2.0-77949515006", "pmid"=>"19997617", "sgr"=>"77949515006"}, "id"=>"e0b1b381-ba62-3a78-92ef-e36f4999b228", "abstract"=>"There is a desperate need for effective therapies to fight chronic viral infections. The immune response is normally fastidious at controlling the majority of viral infections and a therapeutic strategy aimed at reestablishing immune control represents a potentially powerful approach towards treating persistent viral infections. We examined the potential of genetically programming human hematopoietic stem cells to generate mature CD8+ cytotoxic T lymphocytes that express a molecularly cloned, \"transgenic\" human anti-HIV T cell receptor (TCR). Anti-HIV TCR transduction of human hematopoietic stem cells directed the maturation of a large population of polyfunctional, HIV-specific CD8+ cells capable of recognizing and killing viral antigen-presenting cells. Thus, through this proof-of-concept we propose that genetic engineering of human hematopoietic stem cells will allow the tailoring of effector T cell responses to fight HIV infection or other diseases that are characterized by the loss of immune control.", "link"=>"http://www.mendeley.com/research/engineering-antigenspecific-t-cells-genetically-modified-human-hematopoietic-stem-cells-immunodefici-2", "reader_count"=>14, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>2, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>2, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>3, "Other"=>1, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>9, "Medicine and Dentistry"=>1, "Physics and Astronomy"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}}, "reader_count_by_country"=>{"United States"=>1, "Chile"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/873111"], "description"=>"<p>(A). Diagram of the pCCL.PPT.hPGK.tcr1.9.IRES.eGFP lentiviral vector expressing the SL9 specific TCR utilized to genetically manipulate CD34+ cells. LTR, long terminal repeat; hPGK, human phosphoglycerate kinase promoter element; TCRα, alpha chain; 2A peptide, picornavirus 2A self cleaving peptide; TCRβ, beta chain; IRES, internal ribosomal entry site; eGFP, enhanced green fluorescent protein. (B). Transduced Jurkat T cells were analyzed by flow cytometry for expression eGFP and SL9-specific TCR by SL9 tetramer staining 2 days following transduction with the TCR containing lentiviral vector. The numbers represent the frequency of cells expressing the transgenic TCR and eGFP. (C). Purified CD8+ T cells were transduced with the lentiviral vector containing the SL9-specific TCR and assessed by ELISPOT analysis for their ability to express IFN-γ in response to peptide specific stimulation (left) or no peptide treatment (right). The numbers indicate the average number of cells expressing IFN-γ per 10,000 total cells (n = 7 total replicate cultures).</p>", "links"=>[], "tags"=>["hiv", "gag", "sl9", "epitope"], "article_id"=>543517, "categories"=>["Immunology", "Medicine", "Virology", "Computational Biology"], "users"=>["Scott G. Kitchen", "Michael Bennett", "Zoran Galić", "Joanne Kim", "Qing Xu", "Alan Young", "Alexis Lieberman", "Aviva Joseph", "Harris Goldstein", "Hwee Ng", "Otto Yang", "Jerome A. Zack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0008208.g001", "stats"=>{"downloads"=>1, "page_views"=>17, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cloning_and_functional_expression_of_an_HIV_gag_SL9_epitope_specific_TCR_/543517", "title"=>"Cloning and functional expression of an HIV gag SL9 epitope specific TCR.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 03:35:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/873225"], "description"=>"<p>(A). Schematic illustrating the isolation of CD34+ cells from fetal liver by cell sorting (1), transduction of progenitor cells with lentiviral vector containing the SL9-specific TCR (2), and implantation of these cells in the thymic tissue implanted in sub-lethally irradiated SCID-hu mice. Following 3–12 weeks to allow T cell development to occur from HSCs, TCR expression was analyzed and functional assays performed (3). At four (B) and seven (C) weeks following implantation with TCR transduced HSCs, human thymic tissue was biopsied and cells were analyzed by flow cytometry for cell size (forward scatter—denoted “Size”) and SL9 specific tetramer staining. Mock treated mice (top panels) and mice receiving TCR transduced cells (bottom panels) are indicated. The numbers on the left panels illustrate total SL9 tetramer staining cells. SL9-tetramer+ cells were gated and the frequency of cells expressing CD4 and/or CD8 are provided in the right panels.</p>", "links"=>[], "tags"=>["manipulation", "hscs"], "article_id"=>543659, "categories"=>["Immunology", "Medicine", "Virology", "Computational Biology"], "users"=>["Scott G. Kitchen", "Michael Bennett", "Zoran Galić", "Joanne Kim", "Qing Xu", "Alan Young", "Alexis Lieberman", "Aviva Joseph", "Harris Goldstein", "Hwee Ng", "Otto Yang", "Jerome A. Zack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0008208.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Genetic_manipulation_of_HSCs_and_generation_of_thymocytes_/543659", "title"=>"Genetic manipulation of HSCs and generation of thymocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 03:36:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/873603"], "description"=>"<p>(A) Thymic tissues from 2 mice receiving SL9-specific TCR transduced HSCs and 1 mock-treated mouse were biopsied 7 weeks following introduction of HSCs and placed into culture with SL9 peptide coated antigen presenting cells for 1 week to allow differentiation from antigen naïve to effector cells. Effector cells were then stimulated with SL9 peptide or medium alone (no peptide) and IFN-γ production was measured by ELISPOT. (B) Cells from SCID-hu mice receiving SL9-specific TCR transduced HSCs were obtained by biopsy following differentiation into thymocytes and activated in culture in the presence of an irradiated SL9-peptide coated HLA-A*0201+ B cell line and allogeneic PBMC feeder cells. Cells from mouse numbers Y09-13 and Y17-7 were then placed in a standard <sup>51</sup>chromium release assay utilizing SL9 peptide coated T2 cells or untreated T2 cells as a control. Graph shows the specific lytic activity of cells at an effector to target cell ratio of 10∶1.</p>", "links"=>[], "tags"=>["responses", "hiv-specific", "tcr", "transgenic"], "article_id"=>544035, "categories"=>["Immunology", "Medicine", "Virology", "Computational Biology"], "users"=>["Scott G. Kitchen", "Michael Bennett", "Zoran Galić", "Joanne Kim", "Qing Xu", "Alan Young", "Alexis Lieberman", "Aviva Joseph", "Harris Goldstein", "Hwee Ng", "Otto Yang", "Jerome A. Zack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0008208.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Functional_responses_of_HIV_specific_TCR_transgenic_T_cells_/544035", "title"=>"Functional responses of HIV-specific TCR transgenic T cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 03:38:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/873490"], "description"=>"<p>Thy/Liv implants from mice receiving transduced CD34+ cells were biopsied following development of mature TCR expressing cells and isolated thymocytes cultured in the presence of irradiated feeder cells, an irradiated HLA-A*0201+ B cell line and without (top row) or with (bottom row) the SL9 peptide for one week. Cells were removed and analyzed utilizing polychromatic flow cytometry for the indicated markers. Due to down regulation of TCR following peptide-specific stimulation, lentiviral vector expressing cells were identified by expression of eGFP and were gated and analyzed for CD4 versus CD8 expression (left panels). CD8 T cells were then examined using the indicated gate for expression of differentiation markers (middle and right panels). Percentages indicate cells staining in each quadrant and the numbers in parenthesis indicate the percentage of cells within the CD8+ gate.</p>", "links"=>[], "tags"=>["differentiation", "sl-9", "peptide-specific", "stimulation", "transgenic", "tcr", "expressing"], "article_id"=>543906, "categories"=>["Immunology", "Medicine", "Virology", "Computational Biology"], "users"=>["Scott G. Kitchen", "Michael Bennett", "Zoran Galić", "Joanne Kim", "Qing Xu", "Alan Young", "Alexis Lieberman", "Aviva Joseph", "Harris Goldstein", "Hwee Ng", "Otto Yang", "Jerome A. Zack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0008208.g004", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Phenotypic_CD8_T_cell_differentiation_following_SL_9_peptide_specific_stimulation_of_transgenic_TCR_expressing_thymocytes_/543906", "title"=>"Phenotypic CD8+ T cell differentiation following SL-9 peptide-specific stimulation of transgenic TCR expressing thymocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 03:37:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/873345"], "description"=>"<p>(A) Mock treated mice (upper row) and mice receiving HSCs transduced with the HIV SL9-specific TCR (lower row) were analyzed 7 weeks following transplantation for CD3 and SL9-specific TCR expression by tetramer staining of cells from the thymus (left panels) or spleen (right panels). The frequency of CD3+ and SL9-tetramer+ cells is provided and the values inside the parentheses correspond to the percentage of tetramer positive cells in the human T cell (CD3+) populations. (B) Fetal liver derived CD34+ HSCs transduced with the SL9-TCR containing lentiviral vector were implanted into mice containing either HLA-A*0201+ thymic tissue (top panels) or into mice containing HLA-A*0201- thymic tissue (bottom panels) and the frequency of SL-9 tetramer+ cells assessed 6 weeks following implantation. Size (forward scatter) versus tetramer staining is presented in the left panels and the values inside the parentheses correspond to the percentage of tetramer positive cells. Tetramer expressing cells in the indicated gate were assessed for CD4 and CD8 expression (right panels) and the frequencies of cells expressing each marker are provided.</p>", "links"=>[], "tags"=>["sl9-specific", "tcr", "transduced", "hsc"], "article_id"=>543778, "categories"=>["Immunology", "Medicine", "Virology", "Computational Biology"], "users"=>["Scott G. Kitchen", "Michael Bennett", "Zoran Galić", "Joanne Kim", "Qing Xu", "Alan Young", "Alexis Lieberman", "Aviva Joseph", "Harris Goldstein", "Hwee Ng", "Otto Yang", "Jerome A. Zack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0008208.g003", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_SL9_specific_TCR_transduced_HSC_development_into_T_cells_/543778", "title"=>"Characterization of SL9-specific TCR transduced HSC development into T cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 03:37:14"}

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