A Flow Cytometric Assay to Detect Functional Ganglionic Acetylcholine Receptor Antibodies by Immunomodulation in Autoimmune Autonomic Ganglionopathy

Autoimmune Autonomic Ganglionopathy (AAG) is an uncommon immune-mediated neurological disease that results in failure of autonomic function and is associated with autoantibodies directed against the ganglionic acetylcholine receptor (gnACHR). The antibodies are routinely detected by immunoprecipitat...

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Bibliographic Details
Published in:Frontiers in immunology 2021-06, Vol.12, p.705292-705292
Main Authors: Urriola, Nicolás, Spies, Judith M, Blazek, Katrina, Lang, Bethan, Adelstein, Stephen
Format: Article
Language:English
Subjects:
Area Under Curve
Autoantibodies - blood
Autoantibodies - immunology
autoimmune autonomic ganglionopathy
Autoimmune Diseases of the Nervous System - blood
Autoimmune Diseases of the Nervous System - immunology
Autonomic Nervous System Diseases - blood
Autonomic Nervous System Diseases - immunology
Cell Line, Tumor
diagnostic test
Flow Cytometry - methods
Ganglia, Autonomic - immunology
Humans
immunoassay
Immunology
Immunomodulation
neuroimmunology
Plasma
Receptors, Cholinergic - immunology
ROC Curve
Serum
Single-Blind Method
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Summary:Autoimmune Autonomic Ganglionopathy (AAG) is an uncommon immune-mediated neurological disease that results in failure of autonomic function and is associated with autoantibodies directed against the ganglionic acetylcholine receptor (gnACHR). The antibodies are routinely detected by immunoprecipitation assays, such as radioimmunoassays (RIA), although these assays do not detect all patients with AAG and may yield false positive results. Autoantibodies against the gnACHR exert pathology by receptor modulation. Flow cytometric analysis is able to determine if this has occurred, in contrast to the assays in current use that rely on immunoprecipitation. Here, we describe the first high-throughput, non-radioactive flow cytometric assay to determine autoantibody mediated gnACHR immunomodulation. Previously identified gnACHR antibody seronegative and seropositive sera samples (RIA confirmed) were blinded and obtained from the Oxford Neuroimmunology group along with samples collected locally from patients with or without AAG. All samples were assessed for the ability to cause gnACHR immunomodulation utilizing the prototypical gnACHR expressing cell line, IMR-32. Decision limits were calculated from healthy controls, and Receiver Operating Characteristic (ROC) curves were constructed after unblinding all samples. One hundred and ninety serum samples were analyzed; all 182 expected negative samples (from healthy controls, autonomic disorders not thought to be AAG, other neurological disorders without autonomic dysfunction and patients with Systemic Lupus Erythematosus) were negative for immunomodulation (
ISSN:1664-3224
1664-3224
DOI:10.3389/fimmu.2021.705292