See the published version of this article at Molecular Neurodegeneration.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Methodology
David Gate
Stanford University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-0481-9657
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Molecular 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 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.
Keywords
cerebrospinal fluid cells, CSF, intrathecal cells, neurodegeneration, adaptive immunity, T cells, T cell receptor (TCR), antigen
Figure 1
Figure 2
Figure 3
This is a list of supplementary files associated with this preprint. Click to download.
Loading...
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Version 2
Posted 04 Jan, 2021
Published
On 22 Jan, 2021
No community comments so far
Editorial decision: Accept
On 30 Dec, 2020
Review #2 received
Received 26 Dec, 2020
Review #1 received
Received 26 Dec, 2020
Review #3 received
Received 21 Dec, 2020
Reviewer #3 agreed
On 20 Dec, 2020
Reviewer #2 agreed
On 17 Dec, 2020
Reviewers invited
Invitations sent on 17 Dec, 2020
Reviewer #1 agreed
On 17 Dec, 2020
Editor assigned
On 16 Dec, 2020
Submission checks complete
On 16 Dec, 2020
Editor invited
On 16 Dec, 2020
No comments provided
Editorial decision: Major revision
On 19 Oct, 2020
Review #3 received
Received 05 Oct, 2020
Review #2 received
Received 05 Oct, 2020
Reviewer #3 agreed
On 25 Sep, 2020
Reviewer #2 agreed
On 24 Sep, 2020
Review #1 received
Received 02 Sep, 2020
Reviewer #1 agreed
On 24 Aug, 2020
Reviewers invited
Invitations sent on 15 Aug, 2020
Editor assigned
On 06 Aug, 2020
Submission checks complete
On 05 Aug, 2020
Editor invited
On 05 Aug, 2020
First submitted
On 29 Jul, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Neurology
Learn more about our company.
This preprint is under consideration at Molecular Neurodegeneration. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Methodology
David Gate
Stanford University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-0481-9657
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Version 2
Posted 04 Jan, 2021
Published
On 22 Jan, 2021
No community comments so far
Editorial decision: Accept
On 30 Dec, 2020
Review #2 received
Received 26 Dec, 2020
Review #1 received
Received 26 Dec, 2020
Review #3 received
Received 21 Dec, 2020
Reviewer #3 agreed
On 20 Dec, 2020
Reviewer #2 agreed
On 17 Dec, 2020
Reviewers invited
Invitations sent on 17 Dec, 2020
Reviewer #1 agreed
On 17 Dec, 2020
Editor assigned
On 16 Dec, 2020
Submission checks complete
On 16 Dec, 2020
Editor invited
On 16 Dec, 2020
No comments provided
Editorial decision: Major revision
On 19 Oct, 2020
Review #3 received
Received 05 Oct, 2020
Review #2 received
Received 05 Oct, 2020
Reviewer #3 agreed
On 25 Sep, 2020
Reviewer #2 agreed
On 24 Sep, 2020
Review #1 received
Received 02 Sep, 2020
Reviewer #1 agreed
On 24 Aug, 2020
Reviewers invited
Invitations sent on 15 Aug, 2020
Editor assigned
On 06 Aug, 2020
Submission checks complete
On 05 Aug, 2020
Editor invited
On 05 Aug, 2020
First submitted
On 29 Jul, 2020
Subject Areas
Neurology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Molecular 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 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.
Figure 1
Figure 2
Figure 3
Loading...