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@PHDTHESIS{Kreye:144861,
      author       = {Kreye, Jakob},
      title        = {{R}epertoireanalyse monoklonaler {A}ntikörper des
                      {L}iquors von {P}atienten mit
                      anti-{NMDA}-{R}ezeptor-{E}nzephalitis},
      school       = {Charité Universitätsmedizin Berlin},
      type         = {Habilitationsschrift},
      publisher    = {Charité - Universitätsmedizin Berlin},
      reportid     = {DZNE-2020-00286},
      pages        = {28},
      year         = {2017},
      note         = {Habilitationsschrift, Charité Universitätsmedizin Berlin,
                      2017},
      abstract     = {The anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis
                      is the most common form of autoimmune mediated brain
                      inflammation. Typically patients initially develop psychotic
                      changes such as substantive thought disorders and
                      hallucinations, followed by neurological symptoms like
                      impaired consciousness, dyskinesia, seizures, autonomic
                      dysregulations and central hypoventilation. The detection of
                      antibodies against the NR1 subunit of the NMDAR in the
                      cerebrospinal fluid (CSF) is the most important diagnostic
                      criterion. Previous works with extracted whole
                      immunoglobulins from patients‘ CSF and serum have shown
                      synaptic dysfunction in vitro and in vivo. Thereby an
                      influence of further possible autoantibodies could not be
                      eliminated and the proof of pathogenicity of NR1-specific
                      antibodies was pending. Therefore we generated monoclonal
                      recombinant human antibodies from CSF samples from eight
                      patients with NMDAR encephalitis. We isolated single memory
                      B cells and plasma cells via flow cytometry. The variable
                      gene regions coding the heavy and light immunoglobulin
                      chains were amplified by polymerase chain reaction. These
                      were cloned into vectors, containing the constant gene
                      sequence of human antibodies. We co-transfected human
                      embryonic kidney (HEK) cells with the respective pair of
                      heavy and light immunoglobulin chains and harvested
                      monoclonal antibodies from the culture medium. Six percent
                      of the generated monoclonal recombinant human antibodies
                      showed reactivity to NR1-transfected HEK cells, likewise the
                      typical NR1-specific staining pattern on hippocampal neurons
                      and mouse brain sections. All NR1-specific antibodies were
                      from the IgG isotype and have been isolated from plasma
                      cells as well as memory B cells. Last-mentioned ones might
                      be clinically relevant as possible origin of future
                      relapses. Several of the NR1-specific antibodies showed
                      clonal expansion with 100 percent identical clones and few
                      somatic hypermutations, indicating a recent peripheral
                      immune reaction. Furthermore we found three completely
                      unmutated NR1-specific antibodies („naturally occurring
                      antibodies“), suggesting incomplete immune tolerance
                      against NR1 in these patients. Incubating hippocampal
                      neurons with monoclonal NR1-specific antibodies caused a
                      reduction of the NMDAR density and NMDAR-specific currents.
                      After intravenous injections into mice, antibody enrichment
                      in the hippocampus and cerebellum was detected. More than 95
                      percent of the non- NR1-binding antibodies showed specific
                      reactivity on mouse brain sections, amongst others to
                      neuronal surfaces, glia cells and endothelium. With these
                      data we proved that monoclonal NR1-specific antibodies alone
                      are neurotoxic. Therewith, any detection of these
                      autoantibodies needs to be considered as a risk factor for
                      neuro-psychiatric symptoms. Future work will clarify whether
                      the presence of different NR1-specific antibodies is of
                      prognostic relevance and whether further autoantibodies
                      contribute to the pathophysiology.},
      keywords     = {anti-NMDA-receptor-Encephalitis (Other) / monoclonal
                      antibodies (Other) / autoantibodies (Other) / cerebrospinal
                      fluid (Other) / electrophysiology (Other) / 600 Technik,
                      Medizin, angewandte Wissenschaften::610 Medizin und
                      Gesundheit (Other)},
      cin          = {AG Prüß},
      cid          = {I:(DE-2719)1810003},
      pnm          = {342 - Disease Mechanisms and Model Systems (POF3-342)},
      pid          = {G:(DE-HGF)POF3-342},
      typ          = {PUB:(DE-HGF)13},
      urn          = {urn:nbn:de:kobv:188-fudissthesis000000105850-6},
      doi          = {10.17169/REFUBIUM-10947},
      url          = {https://pub.dzne.de/record/144861},
}