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Methods to investigate intrathecal adaptive immunity in neurodegeneration

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 patho...

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Autores principales: Oh, Hamilton, Leventhal, Olivia, Channappa, Divya, Henderson, Victor W., Wyss-Coray, Tony, Lehallier, Benoit, Gate, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7824942/
https://www.ncbi.nlm.nih.gov/pubmed/33482851
http://dx.doi.org/10.1186/s13024-021-00423-w
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author Oh, Hamilton
Leventhal, Olivia
Channappa, Divya
Henderson, Victor W.
Wyss-Coray, Tony
Lehallier, Benoit
Gate, David
author_facet Oh, Hamilton
Leventhal, Olivia
Channappa, Divya
Henderson, Victor W.
Wyss-Coray, Tony
Lehallier, Benoit
Gate, David
author_sort Oh, Hamilton
collection PubMed
description 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13024-021-00423-w.
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spelling pubmed-78249422021-01-25 Methods to investigate intrathecal adaptive immunity in neurodegeneration Oh, Hamilton Leventhal, Olivia Channappa, Divya Henderson, Victor W. Wyss-Coray, Tony Lehallier, Benoit Gate, David Mol Neurodegener Methodology 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13024-021-00423-w. BioMed Central 2021-01-22 /pmc/articles/PMC7824942/ /pubmed/33482851 http://dx.doi.org/10.1186/s13024-021-00423-w Text en © The Author(s) 2021 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Methodology
Oh, Hamilton
Leventhal, Olivia
Channappa, Divya
Henderson, Victor W.
Wyss-Coray, Tony
Lehallier, Benoit
Gate, David
Methods to investigate intrathecal adaptive immunity in neurodegeneration
title 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_short Methods to investigate intrathecal adaptive immunity in neurodegeneration
title_sort methods to investigate intrathecal adaptive immunity in neurodegeneration
topic Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7824942/
https://www.ncbi.nlm.nih.gov/pubmed/33482851
http://dx.doi.org/10.1186/s13024-021-00423-w
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