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A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis

Cell-based assays (CBAs) and radioimmunoprecipitation assay (RIPA) are the most sensitive methods for identifying anti-acetylcholine receptor (AChR) antibody in myasthenia gravis (MG). But CBAs are limited in clinical practice by transient transfection. We established a stable cell line (KL525) expr...

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Autores principales: Cai, Yu, Han, Lu, Zhu, Desheng, Peng, Jing, Li, Jianping, Ding, Jie, Luo, Jiaying, Hong, Ronghua, Wang, Kan, Wan, Wenbin, Xie, Chong, Zhou, Xiajun, Zhang, Ying, Hao, Yong, Guan, Yangtai
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8297518/
https://www.ncbi.nlm.nih.gov/pubmed/34305897
http://dx.doi.org/10.3389/fimmu.2021.666046
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author Cai, Yu
Han, Lu
Zhu, Desheng
Peng, Jing
Li, Jianping
Ding, Jie
Luo, Jiaying
Hong, Ronghua
Wang, Kan
Wan, Wenbin
Xie, Chong
Zhou, Xiajun
Zhang, Ying
Hao, Yong
Guan, Yangtai
author_facet Cai, Yu
Han, Lu
Zhu, Desheng
Peng, Jing
Li, Jianping
Ding, Jie
Luo, Jiaying
Hong, Ronghua
Wang, Kan
Wan, Wenbin
Xie, Chong
Zhou, Xiajun
Zhang, Ying
Hao, Yong
Guan, Yangtai
author_sort Cai, Yu
collection PubMed
description Cell-based assays (CBAs) and radioimmunoprecipitation assay (RIPA) are the most sensitive methods for identifying anti-acetylcholine receptor (AChR) antibody in myasthenia gravis (MG). But CBAs are limited in clinical practice by transient transfection. We established a stable cell line (KL525) expressing clustered AChR by infecting HEK 293T cells with dual lentiviral vectors expressing the genes encoding the human AChR α1, β1, δ, ϵ and the clustering protein rapsyn. We verified the stable expression of human clustered AChR by immunofluorescence, immunoblotting, and real-time PCR. Fluorescence-activated cell sorting (FACS) was used to detect anti-AChR antibodies in 103 MG patients and 58 healthy individuals. The positive results of MG patients reported by the KL525 was 80.6% (83/103), 29.1% higher than the 51.4% (53/103) of RIPA. 58 healthy individuals tested by both the KL525 CBA and RIPA were all negative. In summary, the stable expression of clustered AChR in our cell line makes it highly sensitive and advantageous for broad clinical application in CBAs.
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spelling pubmed-82975182021-07-23 A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis Cai, Yu Han, Lu Zhu, Desheng Peng, Jing Li, Jianping Ding, Jie Luo, Jiaying Hong, Ronghua Wang, Kan Wan, Wenbin Xie, Chong Zhou, Xiajun Zhang, Ying Hao, Yong Guan, Yangtai Front Immunol Immunology Cell-based assays (CBAs) and radioimmunoprecipitation assay (RIPA) are the most sensitive methods for identifying anti-acetylcholine receptor (AChR) antibody in myasthenia gravis (MG). But CBAs are limited in clinical practice by transient transfection. We established a stable cell line (KL525) expressing clustered AChR by infecting HEK 293T cells with dual lentiviral vectors expressing the genes encoding the human AChR α1, β1, δ, ϵ and the clustering protein rapsyn. We verified the stable expression of human clustered AChR by immunofluorescence, immunoblotting, and real-time PCR. Fluorescence-activated cell sorting (FACS) was used to detect anti-AChR antibodies in 103 MG patients and 58 healthy individuals. The positive results of MG patients reported by the KL525 was 80.6% (83/103), 29.1% higher than the 51.4% (53/103) of RIPA. 58 healthy individuals tested by both the KL525 CBA and RIPA were all negative. In summary, the stable expression of clustered AChR in our cell line makes it highly sensitive and advantageous for broad clinical application in CBAs. Frontiers Media S.A. 2021-07-08 /pmc/articles/PMC8297518/ /pubmed/34305897 http://dx.doi.org/10.3389/fimmu.2021.666046 Text en Copyright © 2021 Cai, Han, Zhu, Peng, Li, Ding, Luo, Hong, Wang, Wan, Xie, Zhou, Zhang, Hao and Guan https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Cai, Yu
Han, Lu
Zhu, Desheng
Peng, Jing
Li, Jianping
Ding, Jie
Luo, Jiaying
Hong, Ronghua
Wang, Kan
Wan, Wenbin
Xie, Chong
Zhou, Xiajun
Zhang, Ying
Hao, Yong
Guan, Yangtai
A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis
title A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis
title_full A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis
title_fullStr A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis
title_full_unstemmed A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis
title_short A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis
title_sort stable cell line expressing clustered achr: a novel cell-based assay for anti-achr antibody detection in myasthenia gravis
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8297518/
https://www.ncbi.nlm.nih.gov/pubmed/34305897
http://dx.doi.org/10.3389/fimmu.2021.666046
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