Peripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single-cell transcriptomics
Abstract Background Myasthenia gravis (MG) is the most prevalent autoimmune disorder affecting the neuromuscular junction. A rapid deterioration in respiratory muscle can lead to a myasthenic crisis (MC), which represents a life-threatening condition with high mortality in MG. Multiple CD4+ T subset...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-08-01
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| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12967-023-04421-y |
| _version_ | 1797451705104728064 |
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| author | Huahua Zhong Xiao Huan Rui Zhao Manqiqige Su Chong Yan Jie Song Jianying Xi Chongbo Zhao Feifei Luo Sushan Luo |
| author_facet | Huahua Zhong Xiao Huan Rui Zhao Manqiqige Su Chong Yan Jie Song Jianying Xi Chongbo Zhao Feifei Luo Sushan Luo |
| author_sort | Huahua Zhong |
| collection | DOAJ |
| description | Abstract Background Myasthenia gravis (MG) is the most prevalent autoimmune disorder affecting the neuromuscular junction. A rapid deterioration in respiratory muscle can lead to a myasthenic crisis (MC), which represents a life-threatening condition with high mortality in MG. Multiple CD4+ T subsets and hypercytokinemia have been identified in the peripheral pro-inflammatory milieu during the crisis. However, the pathogenesis is complicated due to the many types of cells involved, leaving the underlying mechanism largely unexplored. Methods We conducted single-cell transcriptomic and immune repertoire sequencing on 33,577 peripheral blood mononuclear cells (PBMCs) from two acetylcholine receptor antibody-positive (AChR +) MG patients during MC and again three months post-MC. We followed the Scanpy workflow for quality control, dimension reduction, and clustering of the single-cell data. Subsequently, we annotated high-resolution cell types utilizing transfer-learning models derived from publicly available single-cell immune datasets. RNA velocity calculations from unspliced and spliced mRNAs were applied to infer cellular state progression. We analyzed cell communication and MG-relevant cytokines and chemokines to identify potential inflammation initiators. Results We identified a unique subset of monocytes, termed monocytes 3 (FCGR3B+ monocytes), which exhibited significant differential expression of pro-inflammatory signaling pathways during and after the crisis. In line with the activated innate immune state indicated by MC, a high neutrophil–lymphocyte ratio (NLR) was confirmed in an additional 22 AChR + MC patients in subsequent hemogram analysis and was associated with MG-relevant clinical scores. Furthermore, oligoclonal expansions were identified in age-associated B cells exhibiting high autoimmune activity, and in CD4+ and CD8+ T cells demonstrating persistent T exhaustion. Conclusions In summary, our integrated analysis of single-cell transcriptomics and TCR/BCR sequencing has underscored the role of innate immune activation which is associated with hypercytokinemia in MC. The identification of a specific monocyte cluster that dominates the peripheral immune profile may provide some hints into the etiology and pathology of MC. However, future functional studies are required to explore causality. |
| first_indexed | 2024-03-09T14:58:21Z |
| format | Article |
| id | doaj.art-a4239e1daecf4466ae4bb09be2645a32 |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-03-09T14:58:21Z |
| publishDate | 2023-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-a4239e1daecf4466ae4bb09be2645a322023-11-26T14:04:49ZengBMCJournal of Translational Medicine1479-58762023-08-0121111410.1186/s12967-023-04421-yPeripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single-cell transcriptomicsHuahua Zhong0Xiao Huan1Rui Zhao2Manqiqige Su3Chong Yan4Jie Song5Jianying Xi6Chongbo Zhao7Feifei Luo8Sushan Luo9Huashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityHuashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityHuashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityHuashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityHuashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityHuashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityHuashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityHuashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityDepartment of Digestive Diseases, Huashan Hospital, Fudan UniversityHuashan Rare Disease Center and Department of Neurology, Huashan Hospital, Shanghai Medical College, National Center for Neurological Disorders, Fudan UniversityAbstract Background Myasthenia gravis (MG) is the most prevalent autoimmune disorder affecting the neuromuscular junction. A rapid deterioration in respiratory muscle can lead to a myasthenic crisis (MC), which represents a life-threatening condition with high mortality in MG. Multiple CD4+ T subsets and hypercytokinemia have been identified in the peripheral pro-inflammatory milieu during the crisis. However, the pathogenesis is complicated due to the many types of cells involved, leaving the underlying mechanism largely unexplored. Methods We conducted single-cell transcriptomic and immune repertoire sequencing on 33,577 peripheral blood mononuclear cells (PBMCs) from two acetylcholine receptor antibody-positive (AChR +) MG patients during MC and again three months post-MC. We followed the Scanpy workflow for quality control, dimension reduction, and clustering of the single-cell data. Subsequently, we annotated high-resolution cell types utilizing transfer-learning models derived from publicly available single-cell immune datasets. RNA velocity calculations from unspliced and spliced mRNAs were applied to infer cellular state progression. We analyzed cell communication and MG-relevant cytokines and chemokines to identify potential inflammation initiators. Results We identified a unique subset of monocytes, termed monocytes 3 (FCGR3B+ monocytes), which exhibited significant differential expression of pro-inflammatory signaling pathways during and after the crisis. In line with the activated innate immune state indicated by MC, a high neutrophil–lymphocyte ratio (NLR) was confirmed in an additional 22 AChR + MC patients in subsequent hemogram analysis and was associated with MG-relevant clinical scores. Furthermore, oligoclonal expansions were identified in age-associated B cells exhibiting high autoimmune activity, and in CD4+ and CD8+ T cells demonstrating persistent T exhaustion. Conclusions In summary, our integrated analysis of single-cell transcriptomics and TCR/BCR sequencing has underscored the role of innate immune activation which is associated with hypercytokinemia in MC. The identification of a specific monocyte cluster that dominates the peripheral immune profile may provide some hints into the etiology and pathology of MC. However, future functional studies are required to explore causality.https://doi.org/10.1186/s12967-023-04421-yMyasthenia gravisMyasthenic crisisSingle-cell sequencingInnate immunityMonocyte |
| spellingShingle | Huahua Zhong Xiao Huan Rui Zhao Manqiqige Su Chong Yan Jie Song Jianying Xi Chongbo Zhao Feifei Luo Sushan Luo Peripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single-cell transcriptomics Journal of Translational Medicine Myasthenia gravis Myasthenic crisis Single-cell sequencing Innate immunity Monocyte |
| title | Peripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single-cell transcriptomics |
| title_full | Peripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single-cell transcriptomics |
| title_fullStr | Peripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single-cell transcriptomics |
| title_full_unstemmed | Peripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single-cell transcriptomics |
| title_short | Peripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single-cell transcriptomics |
| title_sort | peripheral immune landscape for hypercytokinemia in myasthenic crisis utilizing single cell transcriptomics |
| topic | Myasthenia gravis Myasthenic crisis Single-cell sequencing Innate immunity Monocyte |
| url | https://doi.org/10.1186/s12967-023-04421-y |
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