Methods to investigate intrathecal adaptive immunity in neurodegeneration
Abstract Background Cerebrospinal fluid (CSF) provides basic mechanical and immunological protection to the brain. Historically, analysis of CSF has focused on protein changes, yet recent studies have shed light on cellular alterations. Evidence now exists for involvement of intrathecal T cells in t...
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doaj-bbab4b636c0349cbab71e9dfa8bcee312021-01-24T12:23:12ZengBMCMolecular Neurodegeneration1750-13262021-01-0116111110.1186/s13024-021-00423-wMethods to investigate intrathecal adaptive immunity in neurodegenerationHamilton Oh0Olivia Leventhal1Divya Channappa2Victor W. Henderson3Tony Wyss-Coray4Benoit Lehallier5David Gate6Department of Neurology and Neurological Sciences, Stanford University School of MedicineDepartment of Neurology and Neurological Sciences, Stanford University School of MedicineDepartment of Neurology and Neurological Sciences, Stanford University School of MedicineDepartment of Neurology and Neurological Sciences, Stanford University School of MedicineDepartment of Neurology and Neurological Sciences, Stanford University School of MedicineDepartment of Neurology and Neurological Sciences, Stanford University School of MedicineDepartment of Neurology and Neurological Sciences, Stanford University School of MedicineAbstract Background Cerebrospinal fluid (CSF) provides basic mechanical and immunological protection to the brain. Historically, analysis of CSF has focused on protein changes, yet recent studies have shed light on cellular alterations. Evidence now exists for involvement of intrathecal T cells in the pathobiology of neurodegenerative diseases. However, a standardized method for long-term preservation of CSF immune cells is lacking. Further, the functional role of CSF T cells and their cognate antigens in neurodegenerative diseases are largely unknown. Results We present a method for long-term cryopreservation of CSF immune cells for downstream single cell RNA and T cell receptor sequencing (scRNA-TCRseq) analysis. We observe preservation of CSF immune cells, consisting primarily of memory CD4+ and CD8+ T cells. We then utilize unbiased bioinformatics approaches to quantify and visualize TCR sequence similarity within and between disease groups. By this method, we identify clusters of disease-associated, antigen-specific TCRs from clonally expanded CSF T cells of patients with neurodegenerative diseases. Conclusions Here, we provide a standardized approach for long-term storage of CSF immune cells. Additionally, we present unbiased bioinformatic approaches that will facilitate the discovery of target antigens of clonally expanded T cells in neurodegenerative diseases. These novel methods will help improve our understanding of adaptive immunity in the central nervous system.https://doi.org/10.1186/s13024-021-00423-wCerebrospinal fluid cellsCSFIntrathecal cellsNeurodegenerationAdaptive immunityT cells |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hamilton Oh Olivia Leventhal Divya Channappa Victor W. Henderson Tony Wyss-Coray Benoit Lehallier David Gate |
spellingShingle |
Hamilton Oh Olivia Leventhal Divya Channappa Victor W. Henderson Tony Wyss-Coray Benoit Lehallier David Gate Methods to investigate intrathecal adaptive immunity in neurodegeneration Molecular Neurodegeneration Cerebrospinal fluid cells CSF Intrathecal cells Neurodegeneration Adaptive immunity T cells |
author_facet |
Hamilton Oh Olivia Leventhal Divya Channappa Victor W. Henderson Tony Wyss-Coray Benoit Lehallier David Gate |
author_sort |
Hamilton Oh |
title |
Methods to investigate intrathecal adaptive immunity in neurodegeneration |
title_short |
Methods to investigate intrathecal adaptive immunity in neurodegeneration |
title_full |
Methods to investigate intrathecal adaptive immunity in neurodegeneration |
title_fullStr |
Methods to investigate intrathecal adaptive immunity in neurodegeneration |
title_full_unstemmed |
Methods to investigate intrathecal adaptive immunity in neurodegeneration |
title_sort |
methods to investigate intrathecal adaptive immunity in neurodegeneration |
publisher |
BMC |
series |
Molecular Neurodegeneration |
issn |
1750-1326 |
publishDate |
2021-01-01 |
description |
Abstract Background Cerebrospinal fluid (CSF) provides basic mechanical and immunological protection to the brain. Historically, analysis of CSF has focused on protein changes, yet recent studies have shed light on cellular alterations. Evidence now exists for involvement of intrathecal T cells in the pathobiology of neurodegenerative diseases. However, a standardized method for long-term preservation of CSF immune cells is lacking. Further, the functional role of CSF T cells and their cognate antigens in neurodegenerative diseases are largely unknown. Results We present a method for long-term cryopreservation of CSF immune cells for downstream single cell RNA and T cell receptor sequencing (scRNA-TCRseq) analysis. We observe preservation of CSF immune cells, consisting primarily of memory CD4+ and CD8+ T cells. We then utilize unbiased bioinformatics approaches to quantify and visualize TCR sequence similarity within and between disease groups. By this method, we identify clusters of disease-associated, antigen-specific TCRs from clonally expanded CSF T cells of patients with neurodegenerative diseases. Conclusions Here, we provide a standardized approach for long-term storage of CSF immune cells. Additionally, we present unbiased bioinformatic approaches that will facilitate the discovery of target antigens of clonally expanded T cells in neurodegenerative diseases. These novel methods will help improve our understanding of adaptive immunity in the central nervous system. |
topic |
Cerebrospinal fluid cells CSF Intrathecal cells Neurodegeneration Adaptive immunity T cells |
url |
https://doi.org/10.1186/s13024-021-00423-w |
work_keys_str_mv |
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