Dokument: Single B cell antibody technology and high-throughput immune repertoire sequencing as complementary tools to better understand central nervous system (CNS) inflammation and treatment effects
| Titel: | Single B cell antibody technology and high-throughput immune repertoire sequencing as complementary tools to better understand central nervous system (CNS) inflammation and treatment effects | |||||||
| URL für Lesezeichen: | https://docserv.uni-duesseldorf.de/servlets/DocumentServlet?id=57717 | |||||||
| URN (NBN): | urn:nbn:de:hbz:061-20221018-093321-1 | |||||||
| Kollektion: | Dissertationen | |||||||
| Sprache: | Englisch | |||||||
| Dokumententyp: | Wissenschaftliche Abschlussarbeiten » Dissertation | |||||||
| Medientyp: | Text | |||||||
| Autor: | Barman, Sumanta [Autor] | |||||||
| Dateien: |
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| Beitragende: | Prof. Dr. Goebels, Norbert [Gutachter] Prof. Dr. Lang, Philipp A. [Gutachter] | |||||||
| Dewey Dezimal-Klassifikation: | 500 Naturwissenschaften und Mathematik » 570 Biowissenschaften; Biologie | |||||||
| Beschreibung: | Common denominator of this cumulative dissertation is the interaction between the immune
system and the central nervous system (CNS), with a focus on chronic CNS inflammation. Although the CNS is partially protected from major influences of the periphery by a semi selective blood brain barrier (BBB), activated T and B lymphocytes are able to enter the CNS compartment in health and disease. In chronic CNS inflammation, triggered by infectious agents (e.g. herpes simplex virus (HSV) - encephalitis, Neuroborreliosis) or autoimmune/paraneoplastic immunity (e.g. multiple sclerosis (MS), autoimmune encephalitis (AIE) syndromes) often “oligoclonal immunoglobulins” (also known as “oligoclonal bands”, OCB), produced by CNS resident plasma cells, are detectable by diagnostic isoelectric focusing electrophoresis. While their target specificity has not been conclusively identified in multiple sclerosis, characteristic CSF +/- serum antibody signatures of presumably pathogenic relevance have been described in autoimmune encephalitis. This thesis addressed aspects ranging from pathogenesis to treatment of these diseases. Single B cell antibody technology is a dynamic strategy to develop recombinant monoclonal antibody based on the direct amplification of immunoglobulin heavy chain (IgH) and matching light-chain (IgL) variable region encoding genes from single human B cells. In order to recover the typical intrathecal antibody signature in recombinant form, a human monoclonal antibody SSM5 was cloned and expressed from clonally expanded intrathecal plasma cells from a patient with anti N-Methyl-D-aspartic acid (NMDA) receptor encephalitis. We reproduced NMDAR epitope specificity and key pathogenic features of the human disease in vitro and in vivo. Moreover, we showed that, internalization of the recombinant NMDAR-autoantibody complex into endosomes and lysosomes increases the pH-rhodamine fluorescence. Furthermore, we revealed that reduction of NMDAR-mediated responses in oligodendrocytes is mediated by patient derived recombinant human monoclonal SSM5 antibody. High-throughput immune repertoire sequencing is a powerful approach for characterizing adaptive immune responses. We did immune repertoire analysis to characterize longitudinal evolutions in the clonal compositions of patients’ peripheral lymphocyte repertoires before and during alemtuzumab (LEMTRADA®) treatment, and to better understand the beneficial properties and potential side effects of this drug. Immune repertoire sequencing data showed that alemtuzumab has distinctive effects on T cell and B cell repertoires. Moreover, post-alemtuzumab T and B cell repertoires are more diverse than pre-treatment repertoires predominantly in non-secondary autoimmunity patients, in contrast, secondary autoimmunity patients had reduced T cell repertoire diversity, particularly in post-secondary autoimmunity repertoires. Importantly, secondary autoimmunity patients showed significantly increased persistence and expansion of T cell clones already after the first alemtuzumab infusion and a delayed expansion of memory B cell clones after the second infusion, which provides a novel mechanistic explanation for development of secondary autoimmunity combining current B or T cell-related hypotheses. Additionally, we applied a recombinant humanized monoclonal antibody specific for myelin oligodendrocyte glycoprotein (MOG) to induce, together with complement, demyelination in murine organotypic cerebellar slice cultures (OSC) as an ex vivo model of the immune‐CNS interface. Using transgenic mice expressing GFP in myelin allowed us to investigate the effects of potentially protective (in this case the commonly used immunomodulatory intravenous immunoglobulins (IVIG)) and/or remyelination promoting in OSC by live imaging. | |||||||
| Lizenz: |  Urheberrechtsschutz | |||||||
| Fachbereich / Einrichtung: | Mathematisch- Naturwissenschaftliche Fakultät | |||||||
| Dokument erstellt am: | 18.10.2022 | |||||||
| Dateien geändert am: | 18.10.2022 | |||||||
| Promotionsantrag am: | 23.03.2021 | |||||||
| Datum der Promotion: | 28.09.2021 |
