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 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.
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Posted 04 Jan, 2021
On 22 Jan, 2021
On 30 Dec, 2020
Received 26 Dec, 2020
Received 26 Dec, 2020
Received 21 Dec, 2020
On 20 Dec, 2020
On 17 Dec, 2020
Invitations sent on 17 Dec, 2020
On 17 Dec, 2020
On 16 Dec, 2020
On 16 Dec, 2020
On 16 Dec, 2020
On 19 Oct, 2020
Received 05 Oct, 2020
Received 05 Oct, 2020
On 25 Sep, 2020
On 24 Sep, 2020
Received 02 Sep, 2020
On 24 Aug, 2020
Invitations sent on 15 Aug, 2020
On 06 Aug, 2020
On 05 Aug, 2020
On 05 Aug, 2020
On 29 Jul, 2020
Methods to investigate intrathecal adaptive immunity in neurodegeneration
Posted 04 Jan, 2021
On 22 Jan, 2021
On 30 Dec, 2020
Received 26 Dec, 2020
Received 26 Dec, 2020
Received 21 Dec, 2020
On 20 Dec, 2020
On 17 Dec, 2020
Invitations sent on 17 Dec, 2020
On 17 Dec, 2020
On 16 Dec, 2020
On 16 Dec, 2020
On 16 Dec, 2020
On 19 Oct, 2020
Received 05 Oct, 2020
Received 05 Oct, 2020
On 25 Sep, 2020
On 24 Sep, 2020
Received 02 Sep, 2020
On 24 Aug, 2020
Invitations sent on 15 Aug, 2020
On 06 Aug, 2020
On 05 Aug, 2020
On 05 Aug, 2020
On 29 Jul, 2020
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