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Detection of autoantibody against extracellular epitopes of N-methyl-D-aspartate receptor by cell-based assay.
Neurosci Res. 2011 Nov; 71(3):294-302.NR

Abstract

The concept of anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis, a severe, potentially lethal, treatment-responsive disorder, mediated by autoantibodies against NMDAR was proposed. Because paraneoplastic anti-NMDAR encephalitis has a better prognosis after tumor resection and immunotherapy, rapid quantitative systems for detecting functional autoantibodies against extracellular epitopes of NMDAR are necessary. To detect autoantibodies recognizing extracellular epitopes of NMDAR, we stably expressed mutant NMDAR that decreases Ca(2+) permeability on a heterologous cell surface without any antagonist. Serum and CSF samples from patients were analysed using the cells expressing mutant NMDAR subunits by immunocytochemistry and on-cell Western analysis using live cells stably expressing mutant NMDAR. Furthermore, we were able to express mutant GluRζ1(NR1, GluN1) subunit of NMDAR alone on the cell surface and obtained direct evidence of the presence of autoantibodies recognizing extracellular epitopes of GluRζ1 and the induction of internalization by autoantibodies in serum and CSF from patients. The specificity of on-cell Western analysis was improved at 37°C. The combination of this rapid quantitative assay using our on-cell western analysis, detailed analysis of extracellular epitopes of NMDAR, and internalization assay of NMDAR will be valuable for the diagnosis, evaluation of clinical treatments, and follow-up of anti-NMDAR encephalitis.

Authors+Show Affiliations

Department of Molecular Neuroscience, Graduate School of Innovative Life Science, University of Toyama, Toyama 930-0194, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21839784

Citation

Takano, Shiho, et al. "Detection of Autoantibody Against Extracellular Epitopes of N-methyl-D-aspartate Receptor By Cell-based Assay." Neuroscience Research, vol. 71, no. 3, 2011, pp. 294-302.
Takano S, Takahashi Y, Kishi H, et al. Detection of autoantibody against extracellular epitopes of N-methyl-D-aspartate receptor by cell-based assay. Neurosci Res. 2011;71(3):294-302.
Takano, S., Takahashi, Y., Kishi, H., Taguchi, Y., Takashima, S., Tanaka, K., Muraguchi, A., & Mori, H. (2011). Detection of autoantibody against extracellular epitopes of N-methyl-D-aspartate receptor by cell-based assay. Neuroscience Research, 71(3), 294-302. https://doi.org/10.1016/j.neures.2011.07.1834
Takano S, et al. Detection of Autoantibody Against Extracellular Epitopes of N-methyl-D-aspartate Receptor By Cell-based Assay. Neurosci Res. 2011;71(3):294-302. PubMed PMID: 21839784.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Detection of autoantibody against extracellular epitopes of N-methyl-D-aspartate receptor by cell-based assay. AU - Takano,Shiho, AU - Takahashi,Yukitoshi, AU - Kishi,Hiroyuki, AU - Taguchi,Yoshiharu, AU - Takashima,Shutaro, AU - Tanaka,Kortaro, AU - Muraguchi,Atsushi, AU - Mori,Hisashi, Y1 - 2011/08/05/ PY - 2011/05/18/received PY - 2011/07/12/revised PY - 2011/07/24/accepted PY - 2011/8/16/entrez PY - 2011/8/16/pubmed PY - 2012/6/26/medline SP - 294 EP - 302 JF - Neuroscience research JO - Neurosci Res VL - 71 IS - 3 N2 - The concept of anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis, a severe, potentially lethal, treatment-responsive disorder, mediated by autoantibodies against NMDAR was proposed. Because paraneoplastic anti-NMDAR encephalitis has a better prognosis after tumor resection and immunotherapy, rapid quantitative systems for detecting functional autoantibodies against extracellular epitopes of NMDAR are necessary. To detect autoantibodies recognizing extracellular epitopes of NMDAR, we stably expressed mutant NMDAR that decreases Ca(2+) permeability on a heterologous cell surface without any antagonist. Serum and CSF samples from patients were analysed using the cells expressing mutant NMDAR subunits by immunocytochemistry and on-cell Western analysis using live cells stably expressing mutant NMDAR. Furthermore, we were able to express mutant GluRζ1(NR1, GluN1) subunit of NMDAR alone on the cell surface and obtained direct evidence of the presence of autoantibodies recognizing extracellular epitopes of GluRζ1 and the induction of internalization by autoantibodies in serum and CSF from patients. The specificity of on-cell Western analysis was improved at 37°C. The combination of this rapid quantitative assay using our on-cell western analysis, detailed analysis of extracellular epitopes of NMDAR, and internalization assay of NMDAR will be valuable for the diagnosis, evaluation of clinical treatments, and follow-up of anti-NMDAR encephalitis. SN - 1872-8111 UR - https://www.unboundmedicine.com/medline/citation/21839784/Detection_of_autoantibody_against_extracellular_epitopes_of_N_methyl_D_aspartate_receptor_by_cell_based_assay_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0168-0102(11)02035-9 DB - PRIME DP - Unbound Medicine ER -