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  1. AU="Bogert, N V"
  2. AU="Goswami, Sreetosh"
  3. AU="Saraiva, A A F"
  4. AU=Wickenden Julie A
  5. AU="Darbes, Lynae A"
  6. AU="Ostermann, Philipp"
  7. AU="Qiu, Y Z"
  8. AU="Luo, Yue-Jia"
  9. AU="Tamandl, Dietmar" AU="Tamandl, Dietmar"
  10. AU="Holzer, Timothy R"
  11. AU="Zhou, C L"
  12. AU="Lomax, Tony"

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  1. Artikel ; Online: A novel approach to genetic engineering of T-cell subsets by hematopoietic stem cell infection with a bicistronic lentivirus.

    Bogert, N V / Furkel, J / Din, S / Braren, I / Eckstein, V / Müller, J A / Uhlmann, L / Katus, H A / Konstandin, M H

    Scientific reports

    2020  Band 10, Heft 1, Seite(n) 13740

    Abstract: Lentiviral modification of hematopoietic stem cells (HSCs) paved the way for in vivo experimentation and therapeutic approaches in patients with genetic disease. A disadvantage of this method is the use of a ubiquitous promoter leads not only to genetic ... ...

    Abstract Lentiviral modification of hematopoietic stem cells (HSCs) paved the way for in vivo experimentation and therapeutic approaches in patients with genetic disease. A disadvantage of this method is the use of a ubiquitous promoter leads not only to genetic modification of the leukocyte subset of interest e.g. T-cells, but also all other subsequent leukocyte progeny of the parent HSCs. To overcome this limitation we tested a bicistronic lentivirus, enabling subset specific modifications. Designed novel lentiviral constructs harbor a global promoter (mPGK) regulating mCherry for HSCs selection and a T-cell specific promoter upstream of eGFP. Two T-cell specific promoters were assessed: the distal Lck-(dLck) and the CD3δ-promoter. Transduced HSCs were FACS sorted by mCherry expression and transferred into sublethally irradiated C57/BL6 mice. Successful transplantation and T-cell specific expression of eGFP was monitored by peripheral blood assessment. Furthermore, recruitment response of lentiviral engineered leukocytes to the site of inflammation was tested in a peritonitis model without functional impairment. Our constructed lentivirus enables fast generation of subset specific leukocyte transgenesis as shown in T-cells in vivo and opens new opportunities to modify other HSCs derived subsets in the future.
    Mesh-Begriff(e) Animals ; CD3 Complex/genetics ; Cell Line, Tumor ; Gene Transfer Techniques ; Genetic Engineering/methods ; Genetic Vectors/genetics ; Green Fluorescent Proteins/genetics ; Hematopoietic Stem Cell Transplantation/methods ; Hematopoietic Stem Cells/virology ; Inflammation/genetics ; Inflammation/virology ; Lentivirus/genetics ; Lentivirus Infections/virology ; Leukocytes/physiology ; Leukocytes/virology ; Luminescent Proteins/genetics ; Mice ; Mice, Inbred C57BL ; Promoter Regions, Genetic/genetics ; T-Lymphocyte Subsets/physiology ; T-Lymphocyte Subsets/virology
    Chemische Substanzen CD3 Complex ; CD3delta antigen ; Luminescent Proteins ; Green Fluorescent Proteins (147336-22-9)
    Sprache Englisch
    Erscheinungsdatum 2020-08-13
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-020-70793-6
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  2. Artikel ; Online: Dose/time-dependent modulation of the endothelial function through induction agents: non-depleting versus depleting agents.

    Werner, I / Bogert, N V / Stock, U A / Moritz, A / Beiras-Fernandez, A

    Transplantation proceedings

    2014  Band 46, Heft 9, Seite(n) 2953–2956

    Abstract: Background: Polyclonal anti-thymocyte globulins (ATGs) and anti-CD25 antibodies are agents used for induction of immunosuppression in solid-organ transplantation. We aimed to investigate the effect of different regimens of these immunosuppressive ... ...

    Abstract Background: Polyclonal anti-thymocyte globulins (ATGs) and anti-CD25 antibodies are agents used for induction of immunosuppression in solid-organ transplantation. We aimed to investigate the effect of different regimens of these immunosuppressive induction agents on transendothelial migration of peripheral blood mononuclear cells (PBMC) and evaluated the endothelial apoptosis after treatment.
    Methods: Human microvascular endothelial cells were either activated with tumor necrosis factor-α/interferon-γ or not and further treated with 25 or 125 μg/mL ATG (Thymoglobulin, Sanofi-Aventis, Germany) for 2 hours or 24 hours, or with 5 μg/mL Basiliximab (Simulect, Novartis, Germany) for 2 hours or 24 hours. PBMC were either activated with phytohaemagglutinin (PHA) or not and further treated with 25 or 125 μg/mL ATG or with 5 μg/mL Basiliximab for 2 h and then used for transendothelial migration assays. Apoptosis of endothelial cells was detected by means of Annexin-V staining after 2-hour incubation with either 25 or 125 μg/mL ATG or 5 μg/mL Basiliximab.
    Results: Prophylactic 24-hour administration of ATG to naive endothelial cells without PBMC treatment reduced transendothelial migration. Prophylactic 24-hour administration of ATG and Basiliximab to naive endothelial cells after PBMC treatment with the same agents reduced the transendothelial migration after 24 hours. In both cases, no effect could be observed after 2-hour treatment. Basiliximab but not ATG showed a reduction of transmigration after 2-hour treatment of PBMCs without naive EC treatment. Apoptosis of endothelial cells after treatment increased in both cases, being in case of ATG dose-dependent, increasing from 1.2% after either 25 μg/mL ATG to 8.7% after 125 μg/mL ATG.
    Conclusions: Immunosuppressive induction agents modulate the endothelial activity in a dose- and time-dependent manner. Our results suggest that administration of induction agents over longer time periods could provide a potential benefit regarding endothelial immunomodulation. Increased doses may, however, show a deleterious effect on endothelial survival.
    Mesh-Begriff(e) Antibodies, Monoclonal/pharmacology ; Antibodies, Monoclonal/therapeutic use ; Antilymphocyte Serum/pharmacology ; Antilymphocyte Serum/therapeutic use ; Apoptosis/drug effects ; Dose-Response Relationship, Drug ; Endothelial Cells/drug effects ; Graft Rejection/immunology ; Graft Rejection/prevention & control ; Humans ; Immunosuppressive Agents/pharmacology ; Immunosuppressive Agents/therapeutic use ; Leukocytes, Mononuclear/drug effects ; Recombinant Fusion Proteins/pharmacology ; Recombinant Fusion Proteins/therapeutic use ; Time Factors ; Transendothelial and Transepithelial Migration/drug effects
    Chemische Substanzen Antibodies, Monoclonal ; Antilymphocyte Serum ; Immunosuppressive Agents ; Recombinant Fusion Proteins ; basiliximab (9927MT646M) ; thymoglobulin (D7RD81HE4W)
    Sprache Englisch
    Erscheinungsdatum 2014-11
    Erscheinungsland United States
    Dokumenttyp Comparative Study ; Evaluation Studies ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 82046-5
    ISSN 1873-2623 ; 0041-1345
    ISSN (online) 1873-2623
    ISSN 0041-1345
    DOI 10.1016/j.transproceed.2014.06.055
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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