JAMA Neurol 2015;72:96C99. present at disease onset, with an increase in NMDAR antibody titer with the onset of psychiatric symptoms and cognitive deficits. Conclusion: In demyelinating disorders, NMDAR antibodies are uncommon, even in those with symptoms seen in NMDAR encephalitis. NMDA receptor (NMDAR) encephalitis is characterized by the presence of IgG autoantibodies against NMDAR in serum and CSF Mouse monoclonal to KSHV K8 alpha and affects mainly pediatric and young adult female patients. The first symptoms are frequently psychiatric, followed by seizures, dyskinesias, memory deficits, and speech problems, often leading to autonomic instabilities.1 Autoantibodies against NMDAR are also found in cases of CNS NE 10790 demyelinating diseases such as acute disseminated encephalomyelitis (ADEM),2 optic neuritis (ON),2 neuromyelitis optica spectrum disorders (NMOSDs),3 or multiple sclerosis (MS).4 Recent data point to a possible association of NMDAR antibodies with demyelination in some patients, particularly when IgG antibodies against glial surface antigens aquaporin-4 (AQP4) or myelin oligodendrocyte glycoprotein (MOG) are present.5 This is a unique study to systematically investigate the seroprevalence of NMDAR antibodies in patients with CNS demyelinating diseases. We retrospectively screened a cohort of patients with demyelinating diseases for IgG antibodies against NMDAR, AQP4, and MOG. METHODS Patients and controls. Serum samples for this retrospective case-control study were collected in the Clinical Department of Neurology Innsbruck between 2005 and 2014 and stored at ?80C NE 10790 until use. All patients fulfilled diagnostic criteria as described below. Serum samples from 366 individuals were included: 51 patients with ADEM, 41 patients with NMOSD, 34 patients with clinically isolated syndrome (CIS), and 89 patients with MS. Nine patients with NMDAR encephalitis, 94 patients with other neurologic diseases, and 48 NE 10790 healthy individuals were included as controls. Most of the patients have previously been included in a study on NMDAR antibody assay validation6 and in other studies on MOG and/or AQP4 antibodies.7,8 NMOSDs were diagnosed as described by Wingerchuk et al. in 2007, ADEM was diagnosed according to the criteria of the International Pediatric MS Study Group, and MS and CIS were diagnosed according to the 2005 revisions to the McDonald criteria, as previously described.6,C8 Diagnosis of NMDAR encephalitis was based on clinical assessment (e.g., new onset of neuropsychiatric symptoms) and demonstration of antibodies in serum or CSF, as recommended recently.9 The relative frequency of symptoms suggestive of NMDAR encephalitis was selectively increased in our cohort of patients with demyelinating diseases (figure). Open in a separate window Figure Overview of patients with demyelinating diseases, presence of clinical symptoms frequently associated with NMDAR encephalitis, and antibody statusWe included 215 patients with demyelinating CNS disorders. Of these, 81 presented with seizures and/or behavioral symptoms whereas 134 did not. As expected, all 9 patients with NMDA receptor (NMDAR) encephalitis presented with seizures and/or behavioral symptoms. Ab = antibody; ADEM = acute disseminated encephalomyelitis; AQP4 = aquaporin-4; CIS = clinically isolated syndrome; HC = healthy controls; MOG = myelin oligodendrocyte glycoprotein; MS = multiple sclerosis; NMDAR-E = NMDAR encephalitis; NMOSD = neuromyelitis optica spectrum disorder; OND = other neurologic diseases. Standard protocol approvals, registrations, and patient consents. The present study was approved by the ethical committee of Innsbruck Medical University (study numbers AM3041A and AM4059). All patients or their legal representatives gave written informed consent to the study protocol. Live cell-based assays for antibody detection. For the detection of autoantibodies against NMDAR, AQP4, and MOG we used live HEK293A cells transfected with the respective complementary DNA, as described previously.6,7 The following cutoff values were used: 1:20 for NMDAR antibodies, 1:160 for MOG antibodies, and 1:20 for AQP4 antibodies. To prove the specificity of antibody binding, serum samples were preincubated with HEK293A cells expressing NMDAR or nontransfected HEK293A. One CSF sample was tested for NMDAR antibodies as described above, except CSF was applied undiluted but supplemented with MK-801 (Sigma-Aldrich, St. Louis, MO) and further diluted (1:2, 1:4, etc.) in washing buffer containing MK-801. NMDAR antibody testing with fixed cell-based.