Hippocampus (left panel) and cerebellum (right panel) staining patterns. predominantly affected GABABR (39%), LGI1 (17%) and AMPAR (11%) antibodies. Results were reproduced in a separate cohort (B) of 54?AE patients with LGI1, GABABR or AMPAR antibodies in CSF which were missed in 30% by commercial IIFA. Patients with discordant GABABR antibody results (positive in-house but negative commercial IIFA) were less likely to develop full-blown clinical syndrome; no significant clinical differences were noted for the other antibodies. Overall, NSAb testing by commercial IIFA led to false negative results in a substantial number of patients, mainly those affected by anti-LG1, GABABR or AMPAR encephalitis. If these Naphthoquine phosphate disorders are suspected and commercial IIFA is negative, more comprehensive antibody studies are recommended. strong class=”kwd-title” Keywords: neuronal antibodies, brain immunohistochemistry, diagnostic test, autoimmune encephalitis (AE), immunofluorescent assay Introduction Detection of antibodies to neuronal surface proteins and synaptic receptors is important to establish a definitive diagnosis Naphthoquine phosphate of autoimmune encephalitis (1). Well-characterized clinical syndromes associate with specific antibodies, and previously unrecognized neurological diseases are currently defined by the corresponding neuronal surface antibody, such as anti-NMDAR encephalitis or anti-LGI1 encephalitis, which are the most frequent antibody-mediated encephalitis (2). Commercial diagnostic kits using transfected cells that express the most common neuronal surface antigens are widely available, and allow rapid antibody testing in clinical laboratories (3). Most clinical laboratories worldwide use the same commercial indirect immunofluorescence assay (IIFA) whereas rat brain immunohistochemistry is only performed in a few specialized centers (4). However, there are no studies comparing the performance of commercial IIFA with the combination of rodent brain immunohistochemistry and IIFA as used in the initial description of most neuronal surface antibodies. There is preliminary data suggesting that the sensitivity of commercial IIFA, particularly for LGI antibodies in cerebrospinal fluid (CSF), may be low, and Naphthoquine phosphate false positive results may occur particularly when serum is used at high concentration (4, 5). Moreover, commercial diagnostic kits contain a limited number of antigens (up to 6 specificities) and some less frequent or recently described antigens are not included. For these reasons, the use of commercial IIFA as the only method to diagnose autoimmune encephalitis probably misses the detection of otherwise well-characterized and relevant antibodies. This can have important implications such as overlooking the presence of tumors typically associated with some of the non-detected antibodies, or not giving immunotherapy to patients with unrecognized autoimmune encephalitis. Here, we assessed the diagnostic value and limitations of a commercial kit for the detection of neuronal surface antibodies in the serum and CSF of patients with autoimmune encephalitis. Patients and Methods Patients and Samples We prospectively examined 6213 serum and CSF samples from patients referred to our diagnostic lab for detection of antibodies against neuronal surface antigens from October-2016 to October-2020 (Cohort A). Samples were screened with rat brain immunohistochemistry and results were examined by two independent observers. Samples showing positive immunostaining suggestive of a neuropil antibody were first studied with commercial IIFA (6). Samples that were positive on brain immunohistochemistry (with a pattern of staining suggesting a neuronal surface antibody) but negative on commercial IIFA were later studied with in-house IIFA. Clinical data was reviewed in all cases with available information. To further assess the performance of the commercial IIFA, we retrospectively studied 54 consecutive CSF samples from patients with encephalitis and LGI1 (n=12), AMPAR (n=19) or GABABR (n=23) antibodies confirmed by brain immunohistochemistry and in-house IIFA Rabbit Polyclonal to FZD4 (Cohort B). Rat Brain Immunohistochemistry Tissue immunohistochemistry was performed as previously described (7). Briefly, adult Wistar rats were euthanized in a CO2 chamber and the brain was.
Hippocampus (left panel) and cerebellum (right panel) staining patterns