Goebels
Fakultäten » Medizinische Fakultät » Neurologie, Klinik für » Prof. Dr. Norbert Goebels (ausgetreten) » Goebels
Completed research project
| Title / Titel | Molecular and functional characterisation of human recombinant monoclonal antibodies derived from expanded CSF plasma cell clones of multiple sclerosis patients | ||
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| Abstract (PDF, 14 KB) | |||
| Summary / Zusammenfassung | Multiple slerosis (MS) is the most common inflammatory disease of the central nervous system (CNS). While disease course and severity are unpredictible, MS often lead to serious impairment of neurological functions in the affected individuals, usually young adults. While the cause of MS remains to be determined, a contribution of the immune system to the CNS tissue damage is generally accepted. Clonally expanded immunoglobulins in the cerebrospinal fluid (CSF) represent the most constant immunological phenomenon in multiple sclerosis. Although they are detectable as "oligoclonal bands" (OCB) in more than 95% of patients with MS, their antigen specificity and disease relevance has remained unresolved. OCB are thought to be the products of clonally expanded B and plasma cells in the CSF / CNS compartment of MS patients. Aim of this project is the molecular and functional analysis of the immunoglobulins secreted by clonally expanded B and plasma cells in the CSF of MS patients. For this purpose we have established tools to reconstruct the antigen specificity of individual CSF plasma cells. In preliminary experiments we validated this system on CSF plasma cells from a neuroborreliosis (NB) patient. NB is a CNS disease caused by a known infectious agent (Borrelia burgdorferi). NB can sometimes mimic MS and - just like in MS - in the CSF of NB patients OCB are detectable, but with a known specificity for Borrelia burgdorferi (Bb). Therefore NB is an ideal model disease to verify our experimental system. Starting from single FACS sorted CSF plasma cells we performed single cell RT-PCR to amplify the rearranged immunoglobulin heavy and light chain genes. Matching gene pairs of expanded B- / plasma cell clones were then expressed as fully functional, correctly glycosylated, recombinant human monoclonal antibodies in a eukaryotic expression system. The specificity for Borrelia burgdorferi was confirmed in ELISA assays. Using immunoprecipitation and MALDI-TOF analysis we could identify the Bb p41 antigen as the target antigen of one of the mAbs. Using this system we have now derived a panel of recombinant human monoclonal antibodies (“MS-mAbs“) from expanded CSF plasma cell clones from multiple sclerosis patients. In histological brain sections from MS patients, these antibodies display two distinct, mutually exclusive staining patterns, strongly indicating the specificity of the antibodies for CNS tissue. These monoclonal human recombinant antibodies are ideal tools to search for new potential target antigens and to study the pathogenic relevance of autoantibodies in MS and other autoimmune conditions. Aim of this project is to elucidate the antigen specificity of these human MS-mAbs by a variety of different methods such as immune precipitation / MALDI-TOF analysis and recombinant antigen expression libraries. Additionally, the pathogenic potential of the human MS-mAbs will be evaluated in experimental autoimmune encephalomyelitis (EAE) and in organotypic CNS tissue slice cultures. The identification of the actual target antigen(s) will aid our understanding of MS and increase the chance for a future "specific" immunotherapy. |
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| Publications / Publikationen | S. Kuenzle, H.-C. von Büdingen, M. Meier, M. D. Harrer, E. Urich, B. Becher and N. Goebels (2007): Pathogen specificity and autoimmunity distinct features of antigen driven immune responses in neuroborreliosis. Infection and Immunity 75:3842-3847 | ||
| Project leadership and contacts / Projektleitung und Kontakte |
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| Funding source(s) / Unterstützt durch |
SNF (Personen- und Projektförderung) |
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| Duration of Project / Projektdauer | Jul 2007 to Jun 2010 |