Tracking of Vβ8.2-Positive Encephalitogenic T Cells by Complementarity-Determining Region 3 Spectratyping and Subsequent Southern Blot Hybridization in Lewis Rats after Neuroantigen Sensitization1
Tracking of Vβ8.2-Positive Encephalitogenic T Cells by Complementarity-Determining Region 3 Spectratyping and Subsequent Southern Blot Hybridization in Lewis Rats after Neuroantigen Sensitization1
复制标题
神经抗原致敏后，通过互补决定区 3 光谱分型和随后的 Southern 印迹杂交追踪 Vβ8.2 阳性致脑炎 T 细胞1
作者：
H. Sakuma;K. Kohyama;Y. Jee;Y. Matsumoto
器官特异性自身免疫性疾病中的致病性 T 细胞使用有限数量的 TCR α 链和 β 链。在用髓鞘碱性蛋白免疫诱导Lewis大鼠实验性自身免疫性脑脊髓炎（EAE）中，致脑炎T细胞主要使用Vβ8.2 TCR，并且在整个临床EAE过程中在脊髓中发现了含有EAE特异性互补决定区3（CDR3）序列的Vβ8.2谱型的克隆扩增，DSSYEQYFGPG。在本研究中，我们通过 CDR3 谱型分析和随后与致脑炎 CDR3 序列特异性的探针进行 DNA 杂交，对 Vβ8.2 谱型扩展进行了时间和空间分析，以阐明神经抗原致敏后诱导阶段中致脑炎 T 细胞的动力学。结果表明，早在免疫后第3天，Vβ8.2谱型扩展和/或Southern印迹中的阳性信号首先在区域淋巴结中检测到，并在第6天传播到淋巴器官。由于用PBS灌注免疫大鼠，消除了免疫后第3天的阳性信号，因此早期阶段的大多数Vβ8.2阳性致脑炎T细胞将驻留在淋巴管或血管内。此外，免疫接种后 1、3 和 6 天去除足垫中的引流淋巴结并不能改善临床 EAE。这些发现强烈表明致脑炎 T 细胞在致敏后非常迅速地扩散到全身。在克隆水平上分析致病性 T 细胞为设计有效的免疫疗法提供了有用的信息。
Pathogenic T cells in organ-specific autoimmune diseases use a limited number of TCR α- and β-chains. In experimental autoimmune encephalomyelitis (EAE) induced in Lewis rats by immunization with myelin basic protein, encephalitogenic T cells mainly use Vβ8.2 TCR and clonal expansion of the Vβ8.2 spectratype containing the EAE-specific complementarity-determining region 3 (CDR3) sequence, DSSYEQYFGPG, is found in the spinal cord throughout the course of clinical EAE. In the present study we performed temporal and spatial analyses of Vβ8.2 spectratype expansion by CDR3 spectratyping and subsequent DNA hybridization with a probe specific for the encephalitogenic CDR3 sequence to elucidate the kinetics of encephalitogenic T cells during the induction phase after neuroantigen sensitization. It was demonstrated that Vβ8.2 spectratype expansion and/or the positive signal in Southern blot were first detected in the regional lymph nodes as early as day 3 postimmunization and was disseminated over the lymphoid organs by day 6. Because perfusion of immunized rats with PBS erased the positive signals on day 3 postimmunization, the majority of Vβ8.2-positive encephalitogenic T cells at the very early stage would reside within the lymphatic or blood vessels. Furthermore, removal of the draining lymph node 1, 3, and 6 days after immunization in the foot pad did not ameliorate clinical EAE. These findings strongly suggest that encephalitogenic T cells disseminate throughout the whole body very rapidly after sensitization. Analysis of pathogenic T cells at the clonal level provides useful information for designing effective immunotherapy.
易感 MHC 等位基因（而非背景基因）选择自身免疫 T 细胞反应性。
DOI：
10.1172/jci18337
发表时间：
2003
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
Stratmann,Thomas;Martin-Orozco,Natalia;Mallet-Designe,Valerie;Poirot,Laurent;McGavern,Dorian;Losyev,Grigoriy;Dobbs,CathleenM;Oldstone,MichaelBA;Yoshida,Kenji;Kikutani,Hitoshi;Mathis,Diane;Benoist,Christophe;Haskins,Kathryn;Tey
通讯作者：
Tey