1. Repertoire characterization and validation of gB-specific human IgGs directly cloned from humanized mice vaccinated with dendritic cells and protected against HCMV
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Torsten Witte, Martin Messerle, Andreas Schneider, Johannes Koenig, Sebastian J. Theobald, Michael Mach, Matthias Ballmaier, Simon Danisch, Henning Olbrich, Renata Stripecke, Agnes Bonifacius, Lutz Gieselmann, Constanca Figueiredo, Michael Meyer-Hermann, Meryem S. Ercanoglu, Constantin von Kaisenberg, Frank Klawonn, Britta Eiz-Vesper, Florian Klein, Marija Backovic, Christoph Kreer, Sahamoddin Khailaie, Valery Volk, Hannover Medical School [Hannover] (MHH), German Center for Infection Research - partner site Hannover-Braunschweig (DZIF), University of Cologne, University Hospital of Cologne [Cologne], Helmholtz Centre for Infection Research (HZI), Friedrich-Alexander Universität Erlangen-Nürnberg (FAU), Virologie Structurale - Structural Virology, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), German Center for Infection Research - Partner Site Bonn-Cologne (DZIF), Ostfalia University of Applied Sciences, Technische Universität Braunschweig = Technical University of Braunschweig [Braunschweig], R.S./S.T./V.V./H.O. Hannover: This work was financed by grants of the German Center for Infections Research (DZIF-TTU07.803 and DZIF-TTU07.805 to R.S.), by a research collaboration grant of 'The Jackson Laboratory' and by the German Research Council (DFG/SFB738 Project A6 to R.S. and MM, DFG/REBIRTH Unit 6.4 to R.S.). S.T. received a RegSci Ph.D. fellowship, H.O. received a DZIF-Strucmed fellowship and V.V. received a DAAD/ZIB Ph.D. fellowship. F.K./C.K./ Univ. Cologne: This work was funded by grants from the German Center for Infection Research (DZIF, F.K.), the German Research Foundation (CRC 1279, F.K., CRC 1310, C.K. and F.K., Heisenberg-Program KL2389/2-1, F.K.) and the European Research Council (ERC-StG639961, F.K.). M.M.-H./ S.K./ Braunschweig: S.K. was supported by the German Federal Ministry of Education and Research (BMBF) for the eMED project SYSIMIT and by the Helmholtz-Gemeinschaft, Zukunftsthema 'Immunology and Inflammation' (ZT-0027)., European Project: 639961,H2020,ERC-2014-STG,HIV1ABTHERAPY(2016), BRICS, Braunschweiger Zentrum für Systembiologie, Rebenring 56,38106 Braunschweig, Germany, HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany., and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
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B Cells ,Pathology and Laboratory Medicine ,Antibodies, Viral ,Biochemistry ,MESH: Antibodies, Monoclonal ,MESH: Antibodies, Neutralizing ,Mice ,Animal Cells ,Cytotoxic T cell ,MESH: Animals ,Biology (General) ,Enzyme-Linked Immunoassays ,Immune Response ,MESH: Immunoglobulin G ,0303 health sciences ,030302 biochemistry & molecular biology ,Antibodies, Monoclonal ,3. Good health ,Medical Microbiology ,Viral Pathogens ,MESH: Immunization, Passive ,Antibody ,Blood cells ,QH301-705.5 ,T cell ,Immunology ,Cytotoxic T cells ,Microbiology ,03 medical and health sciences ,Antigen ,Genetics ,Humans ,Antibody-Producing Cells ,Immunoassays ,Molecular Biology ,Microbial Pathogens ,MESH: Humans ,Organisms ,Immunization, Passive ,Proteins ,Correction ,MESH: Cytomegalovirus Infections ,Dendritic Cells ,biochemical phenomena, metabolism, and nutrition ,medicine.disease ,Virology ,Antibodies, Neutralizing ,Animal Studies ,Parasitology ,Immunologic diseases. Allergy ,MESH: Disease Models, Animal ,MESH: Antibodies, Viral ,Human cytomegalovirus ,Physiology ,[SDV]Life Sciences [q-bio] ,Cytomegalovirus ,White Blood Cells ,Cytomegalovirus Vaccines ,Medizinische Fakultät ,Immune Physiology ,Cellular types ,Medicine and Health Sciences ,Antigens, Viral ,Immune System Proteins ,biology ,MESH: Dendritic Cells ,T Cells ,Immune cells ,Animal Models ,medicine.anatomical_structure ,Experimental Organism Systems ,Viruses ,Cytomegalovirus Infections ,Human Cytomegalovirus ,Pathogens ,MESH: Antigens, Viral ,Research Article ,MESH: Cytomegalovirus ,Cell biology ,Herpesviruses ,Somatic hypermutation ,Mouse Models ,Research and Analysis Methods ,Antibodies ,Immune system ,Model Organisms ,MESH: Cytomegalovirus Vaccines ,medicine ,Animals ,ddc:610 ,MESH: Mice ,030304 developmental biology ,Biology and life sciences ,RC581-607 ,Disease Models, Animal ,Immunization ,Immunoglobulin G ,biology.protein ,Immunologic Techniques ,DNA viruses - Abstract
Human cytomegalovirus (HCMV) causes serious complications to immune compromised hosts. Dendritic cells (iDCgB) expressing granulocyte-macrophage colony-stimulating factor, interferon-alpha and HCMV-gB were developed to promote de novo antiviral adaptive responses. Mice reconstituted with a human immune system (HIS) were immunized with iDCgB and challenged with HCMV, resulting into 93% protection. Immunization stimulated the expansion of functional effector memory CD8+ and CD4+ T cells recognizing gB. Machine learning analyses confirmed bone marrow T/CD4+, liver B/IgA+ and spleen B/IgG+ cells as predictive biomarkers of immunization (≈87% accuracy). CD8+ and CD4+ T cell responses against gB were validated. Splenic gB-binding IgM-/IgG+ B cells were sorted and analyzed at a single cell level. iDCgB immunizations elicited human-like IgG responses with a broad usage of various IgG heavy chain V gene segments harboring variable levels of somatic hypermutation. From this search, two gB-binding human monoclonal IgGs were generated that neutralized HCMV infection in vitro. Passive immunization with these antibodies provided proof-of-concept evidence of protection against HCMV infection. This HIS/HCMV in vivo model system supported the validation of novel active and passive immune therapies for future clinical translation., Author summary Human cytomegalovirus (HCMV) is a ubiquitous pathogen. As long as the immune system is functional, T and B cells can control HCMV. Yet, for patients who have debilitated immune functions, HCMV infections and reactivations cause major complications. Vaccines or antibodies to prevent or treat HCMV are not yet approved. Novel animal models for testing new immunization approaches are emerging and are important tools to identify biomedical products with a reasonable chance to work in patients. Here, we used a model based on mice transplanted with human immune cells and infected with a traceable HCMV. We tested a cell vaccine (iDCgB) carrying gB, a potent HCMV antigen. The model showed that iDCgB halted the HCMV infection in more than 90% of the mice. We found that antibodies were key players mediating protection. Using state-of-the-art methods, we were able to use the sequences of the human antibodies generated in the mice to construct and produce monoclonal antibodies in the laboratory. Proof-of-concept experiments indicated that administration of these monoclonal antibodies into mice protected them against HCMV infection. In summary, this humanized mouse model was useful to test a vaccine and to generate and test novel antibodies that can be further developed for human use.
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- 2019