Methods to investigate intrathecal adaptive immunity in neurodegeneration.
Methods to investigate intrathecal adaptive immunity in neurodegeneration.
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DOI：
10.1186/s13024-021-00423-w
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发表时间：
2021-01-22
影响因子：
15.1
通讯作者：
Gate D
中科院分区：
文献类型：
--
作者：
Oh H;Leventhal O;Channappa D;Henderson VW;Wyss-Coray T;Lehallier B;Gate D
脑脊液（CSF）为大脑提供基本的机械和免疫保护。从历史上看，CSF的分析主要集中在蛋白质变化上，但最近的研究揭示了细胞的变化。现在有证据表明鞘内T细胞参与神经退行性疾病的病理生物学。然而，缺乏用于长期保存CSF免疫细胞的标准化方法。此外，CSF T细胞及其同源抗原在神经退行性疾病中的功能作用在很大程度上是未知的。我们提出了一种用于长期冷冻保存CSF免疫细胞以用于下游单细胞RNA和T细胞受体测序（scRNA-TCRseq）分析的方法。我们观察到CSF免疫细胞的保存，主要由记忆性CD 4+和CD 8 + T细胞组成。然后，我们利用无偏的生物信息学方法来量化和可视化疾病组内和疾病组之间的TCR序列相似性。通过这种方法，我们从神经退行性疾病患者的克隆扩增的CSF T细胞中鉴定出疾病相关的抗原特异性TCR簇。在这里，我们提供了一种长期储存CSF免疫细胞的标准化方法。此外，我们提出了公正的生物信息学方法，这将有助于发现克隆扩增的T细胞在神经退行性疾病中的靶抗原。这些新方法将有助于提高我们对中枢神经系统适应性免疫的理解。在线版本包含补充材料，可通过10.1186/s13024-021-00423-w获得。
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. 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. 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. The online version contains supplementary material available at 10.1186/s13024-021-00423-w.
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影响因子：
7.3
作者：
Høglund RA;Lossius A;Johansen JN;Homan J;Benth JŠ;Robins H;Bogen B;Bremel RD;Holmøy T
通讯作者：
Holmøy T
DOI：
10.4049/jimmunol.1302884
发表时间：
2014-03-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Han S;Lin YC;Wu T;Salgado AD;Mexhitaj I;Wuest SC;Romm E;Ohayon J;Goldbach-Mansky R;Vanderver A;Marques A;Toro C;Williamson P;Cortese I;Bielekova B
通讯作者：
Bielekova B
影响因子：
8.6
作者：
Johansen, Jorunn N.;Vartdal, Frode;Holmoy, Trygve
通讯作者：
Holmoy, Trygve
影响因子：
64.8
作者：
Sulzer D;Alcalay RN;Garretti F;Cote L;Kanter E;Agin-Liebes J;Liong C;McMurtrey C;Hildebrand WH;Mao X;Dawson VL;Dawson TM;Oseroff C;Pham J;Sidney J;Dillon MB;Carpenter C;Weiskopf D;Phillips E;Mallal S;Peters B;Frazier A;Lindestam Arlehamn CS;Sette A
通讯作者：
Sette A
影响因子：
5.3
作者：
Salou, Marion;Garcia, Alexandra;Michel, Laure;Gainche-Salmon, Anne;Loussouarn, Delphine;Nicol, Bryan;Guillot, Flora;Hulin, Philippe;Nedellec, Steven;Baron, Daniel;Ramstein, Gerard;Soulillou, Jean-Paul;Brouard, Sophie;Nicot, Arnaud B.;Degauque, Nicolas;Laplaud, David A.
通讯作者：
Laplaud, David A.