Analysis oftarget anfigens in oerebrospinal oligoclonal bands in multiple sclerosis patients

多发性硬化症患者脑脊髓寡克隆带靶抗原分析

• 批准号: 13670626
• 负责人: FUJIHARA Kazuo
• 金额: $ 2.43万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670626/
• 关键词: demyelinating disease; neuroimmunology; neurologic intractable diseasa; multiple sclerosis; oligoclonal band

Cerebrospinal fluid (CSF) oligoclonal lgG band (OS) is an important laboratory finding in multiple sclerosis (MS). Since its clinical implication and target antigens remain to be elucidated, we conducted the following analyses. We applied the sensitive isoelectric focusing (IEF) method to detect OB in MS and found that the frequencies of OB were 67.9% in conventional MS (CMS) and 10% in optic-spinal MS (OSMS). Interestingly, the brain MRI findings in OB-negative CMS patients were often atypical. We studied the CSF lgG epitopes with the phage display method. Common aminoacid sequences were detected in each case, and they were frequently homogenous to herpes viral proteins. However, those sequences in each patient were unique. We analyzed the 3 CSF samples of an OB-positive patient and consistently detected the identical sequences homologous to EB viral proteins. Among them, the sequence homologous to EB viral ZEBRA protein reacted more to CSF than serum in VVestern blot, suggesting the relevance to OB. The GST fusion proteins were synthesized, and the CSF lgG reacted to the fusion protein were precipitated in the GST pull down assay. After the absorption, the CSF was subjected to IEF, but the banding pattern of OB was unchanged. Therefore, each band of OB probably consists of multiple antibodies. The viral IL-1O was not detected in any CSF sample obtained during relapse. Some patients have linear trigeminal root lesions, and these unique lesions are similar to those caused by herpes simplex viral infection in animals. The CSF antibody titers to herpes virus were elevated in one OB-positive patient with such lesion during relapse. Our study demonstrated the pathogenetic significance of OB in MS. The peptide sequences shown to react to CSF lgG in MS in the present study may be an important information to further analyze the target antigens of OB.

脑脊液 (CSF) 寡克隆 IgG 带 (OS) 是多发性硬化症 (MS) 的重要实验室发现。由于其临床意义和靶抗原仍有待阐明，我们进行了以下分析。我们应用灵敏等电聚焦（IEF）方法检测MS中的OB，发现传统MS（CMS）中OB的频率为67.9％，视神经MS（OSMS）中OB的频率为10％。有趣的是，OB 阴性 CMS 患者的脑部 MRI 结果往往不典型。我们用噬菌体展示方法研究了脑脊液IgG表位。在每种情况下都检测到了共同的氨基酸序列，并且它们通常与疱疹病毒蛋白同源。然而，每个患者的这些序列都是独特的。我们分析了 OB 阳性患者的 3 个脑脊液样本，一致检测到与 EB 病毒蛋白同源的相同序列。其中，与EB病毒ZEBRA蛋白同源的序列在VVestern印迹中对CSF的反应比对血清的反应更强，提示与OB的相关性。合成GST融合蛋白，并在GST pull down测定中沉淀与融合蛋白反应的CSF IgG。吸收后，对CSF进行IEF，但OB的条带图案没有变化。因此，OB的每个条带可能由多种抗体组成。在复发期间获得的任何脑脊液样本中均未检测到病毒 IL-1O。部分患者存在线状三叉神经根病变，这些独特的病变与动物单纯疱疹病毒感染引起的病变相似。一名 OB 阳性患者在复发期间出现此类病变，脑脊液针对疱疹病毒的抗体滴度升高。我们的研究证明了 OB 在 MS 中的发病意义。本研究显示与MS中的CSF IgG发生反应的肽序列可能是进一步分析OB靶抗原的重要信息。

项目成果

Feng J: "Immunological and virological effects of interferon-α therapy in HTLV-I associated myelopathy/tropical spastic paraparesis"Ann Neurol. 52(suppl1). S72-S72 (2002)

Feng J：“干扰素-α 治疗对 HTLV-I 相关脊髓病/热带痉挛性截瘫的免疫学和病毒学影响”Ann Neurol 52（suppl1）（2002）。

Fujimori J: "The epitope analysis of CSF oligoclonal IgG in Japanese MS patients by phage display method"Neurology. 56(Suppl 3). A225 (2001)

Fujimori J：“通过噬菌体展示法对日本多发性硬化症患者的脑脊液寡克隆 IgG 表位进行分析”神经病学。

Misu T: "Pure neuromyelitis optica : a clinical, MRI, and immunological study in 10 Japanese patients"Neurology. 56(Suppl 3). A196 (2001)

Misu T：“纯视神经脊髓炎：对 10 名日本患者进行的临床、MRI 和免疫学研究”神经病学。

Hayashi T: "Inflammatory demyelinating disease mimicking malignant glioma"J Nucl Med. (in press). (2002)

Hayashi T：“类似于恶性神经胶质瘤的炎症性脱髓鞘疾病”J Nucl Med。

宮澤イザベル: "Neuromyelitis optica (Devic病)と視神経脊髄型MS"脳と神経. 53. 901-910 (2001)

Isabel Miyazawa：“视神经脊髓炎（Devic 病）和视神经脊髓 MS”《大脑与神经》，53. 901-910 (2001)。