Heard R N, Teutsch S M, Bennetts B H, Lee S D, Deane E M, Stewart G J
Neuroimmunology Unit, Department of Immunology, Westmead Hospital, and Department of Medicine, University of Sydney, Sydney, NSW, Australia.
J Neurol Neurosurg Psychiatry. 1999 Nov;67(5):585-90. doi: 10.1136/jnnp.67.5.585.
There have been many studies reporting restricted patterns of T cell receptor usage in established multiple sclerosis and these have led to clinical trials of immunomodulation directed at deleting clonal T cell populations. The present study aims to test the hypothesis that highly restricted T cell populations are also present in the CSF in the earliest clinical stages of acute demyelinating disease of the CNS.
T cell receptor Vbeta (TCRBV) gene expression was studied in CSF and blood in nine patients with acute optic neuritis within 7 days of onset of symptoms, six patients with an acute relapse of multiple sclerosis, and 13 control subjects. RNA was extracted and cDNA synthesised from unstimulated CSF and blood lymphocytes, and TCRBV gene segments were amplified from the cDNA by polymerase chain reaction (PCR) using 21 family specific primers. PCR products were separated by polyacrylamide gel electrophoresis and detected via a labelled oligonucleotide probe. A semiquantitative analysis of band intensity was performed by laser densitometry.
TCRBV mRNA was detected in the CSF of eight of nine patients with optic neuritis, six of six patients with multiple sclerosis, and five of 13 controls, and was closely correlated with the presence of oligoclonal IgG. Usage of a single TCRBV family was demonstrated in two of nine patients with optic neuritis and two of six patients with multiple sclerosis. The number of TCRBV families expressed in the other patients ranged between 5 and 15 (optic neuritis) and 4 and 17 (multiple sclerosis).
There is a relative lack of restriction of TCRBV usage by CSF lymphocytes in the very earliest stages (<7 days) of acute optic neuritis. This may imply either that multiple sclerosis is not a monoclonal disease even at onset, or that the autoimmune response has widened before the disease becomes clinically evident. This may have important consequences for the design of immune therapies in multiple sclerosis. Further studies are required to determine whether the CSF T cell repertoire at presentation has prognostic importance. Longitudinal studies are required to follow the CSF T cell repertoire from the time of presentation and to determine whether it may have prognostic significance.
已有多项研究报道在确诊的多发性硬化症中存在T细胞受体使用受限模式，这些研究已促成针对消除克隆性T细胞群体的免疫调节临床试验。本研究旨在检验以下假设：在中枢神经系统急性脱髓鞘疾病的最早临床阶段，脑脊液中也存在高度受限的T细胞群体。
对9例症状出现7天内的急性视神经炎患者、6例多发性硬化症急性复发患者和13例对照者的脑脊液和血液中的T细胞受体Vβ（TCRBV）基因表达进行了研究。从未经刺激的脑脊液和血液淋巴细胞中提取RNA并合成cDNA，使用21种家族特异性引物通过聚合酶链反应（PCR）从cDNA中扩增TCRBV基因片段。PCR产物通过聚丙烯酰胺凝胶电泳分离，并通过标记的寡核苷酸探针进行检测。通过激光密度测定法对条带强度进行半定量分析。
在9例视神经炎患者中的8例、6例多发性硬化症患者中的6例和13例对照者中的5例的脑脊液中检测到TCRBV mRNA，且与寡克隆IgG的存在密切相关。9例视神经炎患者中的2例和6例多发性硬化症患者中的2例表现出单一TCRBV家族的使用。其他患者中表达的TCRBV家族数量在5至15个（视神经炎）和4至17个（多发性硬化症）之间。
在急性视神经炎的最早阶段（<7天），脑脊液淋巴细胞对TCRBV的使用相对缺乏限制。这可能意味着即使在发病时多发性硬化症也不是单克隆疾病，或者自身免疫反应在疾病临床显现之前已经扩大。这可能对多发性硬化症免疫治疗的设计产生重要影响。需要进一步研究以确定就诊时脑脊液中的T细胞库是否具有预后重要性。需要进行纵向研究以跟踪从就诊时起的脑脊液T细胞库，并确定其是否可能具有预后意义。
