Abstract
Autoimmune encephalitis (AIE) is a group of disorders in which autoantibodies directed at antigens located on the plasma membrane of neurons induce severe neurological symptoms. In contrast to classical paraneoplastic disorders, AIE patients respond well to immunotherapy. The detection of neuronal surface autoantibodies in patients’ serum or CSF therefore has serious consequences for the patients’ treatment and follow-up and requires the availability of sensitive and specific diagnostic tests. This mini-review provides a guideline for both diagnostic and research laboratories that work on the detection of known surface autoantibodies and/or the identification of novel surface antigens. We discuss the strengths and pitfalls of different techniques for anti-neuronal antibody detection: (1) Immunohistochemistry (IHC) and immunofluorescence on rat/primate brain sections; (2) Immunocytochemistry (ICC) of living cultured hippocampal neurons; and (3) Cell Based Assay (CBA). In addition, we discuss the use of immunoprecipitation and mass spectrometry analysis for the detection of novel neuronal surface antigens, which is a crucial step in further disease classification and the development of novel CBAs.
Highlights
Anti-neuronal autoimmune encephalitis (AIE) is a heterogeneous group of disorders characterized by autoantibodies that are directed at the extracellular domains of antigens in the synaptic or extra-synaptic plasma membrane
In this article we review the advantages and pitfalls of three different techniques for antibody detection: (1) IHC/indirect immunofluorescence (IIF) on adult rat/primate brain slices; (2) ICC on living cultured rat hippocampal neurons; and (3) Cell Based Assay (CBA) for neuronal membrane proteins
In addition we evaluate the use of immunoprecipitation and mass spectrometry analysis for the identification of novel cell-surface antigens
Summary
Anti-neuronal autoimmune encephalitis (AIE) is a heterogeneous group of disorders characterized by autoantibodies that are directed at the extracellular domains of antigens in the synaptic or extra-synaptic plasma membrane. Core methods in PNS and AIE diagnostics are immunohistochemistry (IHC) on rat brain sections and indirect immunofluorescence (IIF) on primate cerebellum sections In these assays all relevant antigens are present and accessible. For the detection of neuronal surface antigens immunocytochemistry (ICC) of primary hippocampal neurons is used These techniques are very useful as initial screening methods, they do not allow for the identification of the exact molecular target of the autoantibodies. All possible antigens are available and accessible, and different brain regions can be assessed This technique has been a core method for the detection of antibodies directed at intracellular antigens in PNS. For cell-surface antigens, the hippocampus is scored for staining of the synapses containing gray matter, termed neuropil (Figure 1A) This neuropil staining is less robust when using the classical PNS pre-treatment of rat brain tissue.
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