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
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
Affiliations
Milita Darguzyte
Institute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Philipp Antczak
Department II of Internal Medicine, Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Daniel Bachurski
Department 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, Germany
Patrick Hoelker
Department 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, Germany
Nima Abedpour
Department 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, Germany
Rahil Gholamipoorfard
Department 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, Germany
Hans A. Schlößer
Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Kerstin Wennhold
Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Martin Thelen
Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Maria A. Garcia-Marquez
Center for Molecular Medicine Cologne (CMMC), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Johannes Koenig
Department of Hematology, Oncology, Hemostasis and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, Germany
Andreas Schneider
Department of Hematology, Oncology, Hemostasis and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, Germany
Tobias Braun
Department of Hematology, Oncology, Hemostasis and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, Germany
Frank Klawonn
Department of Computer Science, Ostfalia University of Applied Sciences, 38302 Wolfenbuettel, Germany
Michael Damrat
Institute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Masudur Rahman
Institute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Jan-Malte Kleid
Institute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Sebastian J. Theobald
Department 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, Germany
Eugen Bauer
Institute of Transfusion Medicine, Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Constantin von Kaisenberg
Department of Obstetrics, Gynecology and Reproductive Medicine, Hannover Medical School, 30625 Hannover, Germany
Steven R. Talbot
Institute for Laboratory Animal Science, Hannover Medical School, 30625 Hannover, Germany
Leonard D. Shultz
The Jackson Laboratory, Bar Harbor, ME 04609, USA
Brian Soper
The Jackson Laboratory, Bar Harbor, ME 04609, USA
Renata Stripecke
Institute for Translational Immune-Oncology, Cancer Research Center Cologne-Essen (CCCE), Faculty of Medicine and University Hospital Cologne, University of Cologne, 50931 Cologne, Germany
Background: Humanized mice transplanted with CD34+ 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+ and CD8+ 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.
