Long-Term Human Immune Reconstitution, T-Cell Development, and Immune Reactivity in Mice Lacking the Murine Major Histocompatibility Complex: Validation with Cellular and Gene Expression Profiles
Background: Humanized mice transplanted with CD34<sup>+</sup> hematopoietic cells (HPCs) are broadly used to study human immune responses and infections in vivo and for testing therapies pre-clinically. However, until now, it was not clear whether interactions between the mouse major his...
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| author | Milita Darguzyte Philipp Antczak Daniel Bachurski Patrick Hoelker Nima Abedpour Rahil Gholamipoorfard Hans A. Schlößer Kerstin Wennhold Martin Thelen Maria A. Garcia-Marquez Johannes Koenig Andreas Schneider Tobias Braun Frank Klawonn Michael Damrat Masudur Rahman Jan-Malte Kleid Sebastian J. Theobald Eugen Bauer Constantin von Kaisenberg Steven R. Talbot Leonard D. Shultz Brian Soper Renata Stripecke |
| author_facet | Milita Darguzyte Philipp Antczak Daniel Bachurski Patrick Hoelker Nima Abedpour Rahil Gholamipoorfard Hans A. Schlößer Kerstin Wennhold Martin Thelen Maria A. Garcia-Marquez Johannes Koenig Andreas Schneider Tobias Braun Frank Klawonn Michael Damrat Masudur Rahman Jan-Malte Kleid Sebastian J. Theobald Eugen Bauer Constantin von Kaisenberg Steven R. Talbot Leonard D. Shultz Brian Soper Renata Stripecke |
| author_sort | Milita Darguzyte |
| collection | DOAJ |
| description | Background: Humanized mice transplanted with CD34<sup>+</sup> hematopoietic cells (HPCs) are broadly used to study human immune responses and infections in vivo and for testing therapies pre-clinically. However, until now, it was not clear whether interactions between the mouse major histocompatibility complexes (MHCs) and/or the human leukocyte antigens (HLAs) were necessary for human T-cell development and immune reactivity. Methods: We evaluated the long-term (20-week) human hematopoiesis and human T-cell development in NOD Scid Gamma (NSG) mice lacking the expression of MHC class I and II (NSG-DKO). Triplicate experiments were performed with HPCs obtained from three donors, and humanization was confirmed in the reference strain NOD Rag Gamma (NRG). Further, we tested whether humanized NSG-DKO mice would respond to a lentiviral vector (LV) systemic delivery of HLA-A*02:01, HLA-DRB1*04:01, human GM-CSF/IFN-α, and the human cytomegalovirus gB antigen. Results: Human immune reconstitution was detectable in peripheral blood from 8 to 20 weeks after the transplantation of NSG-DKO. Human single positive CD4<sup>+</sup> and CD8<sup>+</sup> T-cells were detectable in lymphatic tissues (thymus, bone marrow, and spleen). LV delivery harnessed the detection of lymphocyte subsets in bone marrow (αβ and γδ T-cells and NK cells) and the expression of HLA-DR. Furthermore, RNA sequencing showed that LV delivery increased the expression of different human reactome pathways, such as defense responses to other organisms and viruses. Conclusions: Human T-cell development and reactivity are independent of the expression of murine MHCs in humanized mice. Therefore, humanized NSG-DKO is a promising new model for studying human immune responses, as it abrogates the xenograft mouse MHC interference. |
| format | Article |
| id | doaj-art-633b76dd515d43d3b3abd9316c52d10a |
| institution | OA Journals |
| issn | 2073-4409 |
| language | English |
| publishDate | 2024-10-01 |
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| spelling | doaj-art-633b76dd515d43d3b3abd9316c52d10a2025-08-20T02:11:09ZengMDPI AGCells2073-44092024-10-011320168610.3390/cells13201686Long-Term Human Immune Reconstitution, T-Cell Development, and Immune Reactivity in Mice Lacking the Murine Major Histocompatibility Complex: Validation with Cellular and Gene Expression ProfilesMilita Darguzyte0Philipp Antczak1Daniel Bachurski2Patrick Hoelker3Nima Abedpour4Rahil Gholamipoorfard5Hans A. Schlößer6Kerstin Wennhold7Martin Thelen8Maria A. Garcia-Marquez9Johannes Koenig10Andreas Schneider11Tobias Braun12Frank Klawonn13Michael Damrat14Masudur Rahman15Jan-Malte Kleid16Sebastian J. Theobald17Eugen Bauer18Constantin von Kaisenberg19Steven R. Talbot20Leonard D. Shultz21Brian Soper22Renata Stripecke23Institute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyDepartment II of Internal Medicine, Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyDepartment I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf, Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyDepartment I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf, Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyDepartment I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf, Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyDepartment I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf, Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyCenter for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyCenter for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyCenter for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyCenter for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyDepartment of Hematology, Oncology, Hemostasis and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, GermanyDepartment of Hematology, Oncology, Hemostasis and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, GermanyDepartment of Hematology, Oncology, Hemostasis and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, GermanyDepartment of Computer Science, Ostfalia University of Applied Sciences, 38302 Wolfenbuettel, GermanyInstitute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyInstitute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyInstitute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyDepartment I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Düsseldorf, Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyInstitute of Transfusion Medicine, Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyDepartment of Obstetrics, Gynecology and Reproductive Medicine, Hannover Medical School, 30625 Hannover, GermanyInstitute for Laboratory Animal Science, Hannover Medical School, 30625 Hannover, GermanyThe Jackson Laboratory, Bar Harbor, ME 04609, USAThe Jackson Laboratory, Bar Harbor, ME 04609, USAInstitute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, GermanyBackground: Humanized mice transplanted with CD34<sup>+</sup> hematopoietic cells (HPCs) are broadly used to study human immune responses and infections in vivo and for testing therapies pre-clinically. However, until now, it was not clear whether interactions between the mouse major histocompatibility complexes (MHCs) and/or the human leukocyte antigens (HLAs) were necessary for human T-cell development and immune reactivity. Methods: We evaluated the long-term (20-week) human hematopoiesis and human T-cell development in NOD Scid Gamma (NSG) mice lacking the expression of MHC class I and II (NSG-DKO). Triplicate experiments were performed with HPCs obtained from three donors, and humanization was confirmed in the reference strain NOD Rag Gamma (NRG). Further, we tested whether humanized NSG-DKO mice would respond to a lentiviral vector (LV) systemic delivery of HLA-A*02:01, HLA-DRB1*04:01, human GM-CSF/IFN-α, and the human cytomegalovirus gB antigen. Results: Human immune reconstitution was detectable in peripheral blood from 8 to 20 weeks after the transplantation of NSG-DKO. Human single positive CD4<sup>+</sup> and CD8<sup>+</sup> T-cells were detectable in lymphatic tissues (thymus, bone marrow, and spleen). LV delivery harnessed the detection of lymphocyte subsets in bone marrow (αβ and γδ T-cells and NK cells) and the expression of HLA-DR. Furthermore, RNA sequencing showed that LV delivery increased the expression of different human reactome pathways, such as defense responses to other organisms and viruses. Conclusions: Human T-cell development and reactivity are independent of the expression of murine MHCs in humanized mice. Therefore, humanized NSG-DKO is a promising new model for studying human immune responses, as it abrogates the xenograft mouse MHC interference.https://www.mdpi.com/2073-4409/13/20/1686stem cell transplantationNSGhumanized miceMHCHLAT-cell |
| spellingShingle | Milita Darguzyte Philipp Antczak Daniel Bachurski Patrick Hoelker Nima Abedpour Rahil Gholamipoorfard Hans A. Schlößer Kerstin Wennhold Martin Thelen Maria A. Garcia-Marquez Johannes Koenig Andreas Schneider Tobias Braun Frank Klawonn Michael Damrat Masudur Rahman Jan-Malte Kleid Sebastian J. Theobald Eugen Bauer Constantin von Kaisenberg Steven R. Talbot Leonard D. Shultz Brian Soper Renata Stripecke Long-Term Human Immune Reconstitution, T-Cell Development, and Immune Reactivity in Mice Lacking the Murine Major Histocompatibility Complex: Validation with Cellular and Gene Expression Profiles Cells stem cell transplantation NSG humanized mice MHC HLA T-cell |
| title | Long-Term Human Immune Reconstitution, T-Cell Development, and Immune Reactivity in Mice Lacking the Murine Major Histocompatibility Complex: Validation with Cellular and Gene Expression Profiles |
| title_full | Long-Term Human Immune Reconstitution, T-Cell Development, and Immune Reactivity in Mice Lacking the Murine Major Histocompatibility Complex: Validation with Cellular and Gene Expression Profiles |
| title_fullStr | Long-Term Human Immune Reconstitution, T-Cell Development, and Immune Reactivity in Mice Lacking the Murine Major Histocompatibility Complex: Validation with Cellular and Gene Expression Profiles |
| title_full_unstemmed | Long-Term Human Immune Reconstitution, T-Cell Development, and Immune Reactivity in Mice Lacking the Murine Major Histocompatibility Complex: Validation with Cellular and Gene Expression Profiles |
| title_short | Long-Term Human Immune Reconstitution, T-Cell Development, and Immune Reactivity in Mice Lacking the Murine Major Histocompatibility Complex: Validation with Cellular and Gene Expression Profiles |
| title_sort | long term human immune reconstitution t cell development and immune reactivity in mice lacking the murine major histocompatibility complex validation with cellular and gene expression profiles |
| topic | stem cell transplantation NSG humanized mice MHC HLA T-cell |
| url | https://www.mdpi.com/2073-4409/13/20/1686 |
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