Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized Mice

Antiviral neutralizing antibodies (nAbs) are commonly derived from B cells developed in immunized or infected animals and humans. Fully human antibodies are preferred for clinical use as they are potentially less immunogenic. However, the function of B cells varies depending on their homing pattern...

Full description

Bibliographic Details
Main Authors: Sebastian J. Theobald, Elena Fiestas, Andreas Schneider, Benjamin Ostermann, Simon Danisch, Constantin von Kaisenberg, Jan Rybniker, Wolfgang Hammerschmidt, Reinhard Zeidler, Renata Stripecke
Format: Article
Language:English
Published: MDPI AG 2023-12-01
Series:Cells
Subjects:
Online Access:https://www.mdpi.com/2073-4409/13/1/20
_version_ 1797359042751889408
author Sebastian J. Theobald
Elena Fiestas
Andreas Schneider
Benjamin Ostermann
Simon Danisch
Constantin von Kaisenberg
Jan Rybniker
Wolfgang Hammerschmidt
Reinhard Zeidler
Renata Stripecke
author_facet Sebastian J. Theobald
Elena Fiestas
Andreas Schneider
Benjamin Ostermann
Simon Danisch
Constantin von Kaisenberg
Jan Rybniker
Wolfgang Hammerschmidt
Reinhard Zeidler
Renata Stripecke
author_sort Sebastian J. Theobald
collection DOAJ
description Antiviral neutralizing antibodies (nAbs) are commonly derived from B cells developed in immunized or infected animals and humans. Fully human antibodies are preferred for clinical use as they are potentially less immunogenic. However, the function of B cells varies depending on their homing pattern and an additional hurdle for antibody discovery in humans is the source of human tissues with an immunological microenvironment. Here, we show an efficient method to pharm human antibodies using immortalized B cells recovered from Nod.Rag.Gamma (NRG) mice reconstituting the human immune system (HIS). Humanized HIS mice were immunized either with autologous engineered dendritic cells expressing the human cytomegalovirus gB envelope protein (HCMV-gB) or with Epstein–Barr virus-like particles (EB-VLP). Human B cells recovered from spleen of HIS mice were efficiently immortalized with EBV in vitro. We show that these immortalized B cells secreted human IgGs with neutralization capacities against prototypic HCMV-gB and EBV-gp350. Taken together, we show that HIS mice can be successfully used for the generation and pharming fully human IgGs. This technology can be further explored to generate antibodies against emerging infections for diagnostic or therapeutic purposes.
first_indexed 2024-03-08T15:10:56Z
format Article
id doaj.art-fbf2eb71ec7849e58367a95268336eee
institution Directory Open Access Journal
issn 2073-4409
language English
last_indexed 2024-03-08T15:10:56Z
publishDate 2023-12-01
publisher MDPI AG
record_format Article
series Cells
spelling doaj.art-fbf2eb71ec7849e58367a95268336eee2024-01-10T14:53:13ZengMDPI AGCells2073-44092023-12-011312010.3390/cells13010020Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized MiceSebastian J. Theobald0Elena Fiestas1Andreas Schneider2Benjamin Ostermann3Simon Danisch4Constantin von Kaisenberg5Jan Rybniker6Wolfgang Hammerschmidt7Reinhard Zeidler8Renata Stripecke9Department I of Internal Medicine, University Hospital Cologne, Faculty of Medicine, University of Cologne, 50937 Cologne, GermanyResearch Unit Gene Vectors, Helmholtz Center Munich, German Research Center for Environmental Health, 81377 Munich, GermanyClinic of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, GermanyClinic of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, GermanyClinic of Hematology, Hemostasis, Oncology and Stem Cell Transplantation, Hannover Medical School, 30625 Hannover, GermanyDepartment of Obstetrics, Gynecology and Reproductive Medicine, Hannover Medical School, 30625 Hannover, GermanyDepartment I of Internal Medicine, University Hospital Cologne, Faculty of Medicine, University of Cologne, 50937 Cologne, GermanyResearch Unit Gene Vectors, Helmholtz Center Munich, German Research Center for Environmental Health, 81377 Munich, GermanyGerman Center for Infection Research (DZIF), Partner Site Munich, 81377 Munich, GermanyDepartment I of Internal Medicine, University Hospital Cologne, Faculty of Medicine, University of Cologne, 50937 Cologne, GermanyAntiviral neutralizing antibodies (nAbs) are commonly derived from B cells developed in immunized or infected animals and humans. Fully human antibodies are preferred for clinical use as they are potentially less immunogenic. However, the function of B cells varies depending on their homing pattern and an additional hurdle for antibody discovery in humans is the source of human tissues with an immunological microenvironment. Here, we show an efficient method to pharm human antibodies using immortalized B cells recovered from Nod.Rag.Gamma (NRG) mice reconstituting the human immune system (HIS). Humanized HIS mice were immunized either with autologous engineered dendritic cells expressing the human cytomegalovirus gB envelope protein (HCMV-gB) or with Epstein–Barr virus-like particles (EB-VLP). Human B cells recovered from spleen of HIS mice were efficiently immortalized with EBV in vitro. We show that these immortalized B cells secreted human IgGs with neutralization capacities against prototypic HCMV-gB and EBV-gp350. Taken together, we show that HIS mice can be successfully used for the generation and pharming fully human IgGs. This technology can be further explored to generate antibodies against emerging infections for diagnostic or therapeutic purposes.https://www.mdpi.com/2073-4409/13/1/20HCMVEBVherpesvirushumanized miceantibodiesimmortalization
spellingShingle Sebastian J. Theobald
Elena Fiestas
Andreas Schneider
Benjamin Ostermann
Simon Danisch
Constantin von Kaisenberg
Jan Rybniker
Wolfgang Hammerschmidt
Reinhard Zeidler
Renata Stripecke
Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized Mice
Cells
HCMV
EBV
herpesvirus
humanized mice
antibodies
immortalization
title Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized Mice
title_full Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized Mice
title_fullStr Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized Mice
title_full_unstemmed Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized Mice
title_short Fully Human Herpesvirus-Specific Neutralizing IgG Antibodies Generated by EBV Immortalization of Splenocytes-Derived from Immunized Humanized Mice
title_sort fully human herpesvirus specific neutralizing igg antibodies generated by ebv immortalization of splenocytes derived from immunized humanized mice
topic HCMV
EBV
herpesvirus
humanized mice
antibodies
immortalization
url https://www.mdpi.com/2073-4409/13/1/20
work_keys_str_mv AT sebastianjtheobald fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT elenafiestas fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT andreasschneider fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT benjaminostermann fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT simondanisch fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT constantinvonkaisenberg fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT janrybniker fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT wolfganghammerschmidt fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT reinhardzeidler fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice
AT renatastripecke fullyhumanherpesvirusspecificneutralizingiggantibodiesgeneratedbyebvimmortalizationofsplenocytesderivedfromimmunizedhumanizedmice