Identification of a specific autoantigen in opticospinal form of multiple sclerosis based on the HLA-DPB1 binding motif

基于 HLA-DPB1 结合基序鉴定多发性硬化症视脊髓形式的特异性自身抗原

基本信息

批准号：
12557060
负责人：
KIRA Jun-ichi
金额：
$ 8.64万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

Multiple sclerosis (MS) is currently hypothesized to be mediated by autoreactive T cells against myelin antigens. In Japanese, MS frequently and selectively affects the optic nerve and spinal cord. We have previously reported that the opticospinal form of MS (OS-MS) has been shown to be distinct from the conventional form of MS (C-MS) clinically, immunologically and immunogenetically. Recently, oligodendrocytes have been demonstrated to have plural origins, showing marked differences in distribution depending on their origin. We assumed that the specific autoantigens which exist in the optic nerve and spinal cord may be associated with development of OS-MS lesions.To identify the autoantigens in OS-MS, we made use of an immunoscreening of a human spinal cord cDNA expression library with serum samples obtained from patients with OS-MS. cDNA expression library was prepared from human spinal cord and 5 X 10^5 to 10 X 10^5 phage plaques were immunoscreened with sera obtained from eight OS- … More MS patients. Eleven positive immunoreactive clones were identified by their reactivity to the sera, and their sequences were analyzed. Comparisons of the sequences showed that these clones represented cDNAs from 5 distinct genes (HSP105, Rabaptin-5, NSD1, KIAA1640, KIAA0640). We produced GST-Rabaptin-5 fusion protein and HSP105 protein and evaluated the titer of IgG anti-Rabaptin-5 and anti-HSP105 autoantibody in sera and CSF, using ELISA. We found IgG anti-Rabaptin-5 reactivities were not significantly elevated, however IgG anti-HSP105 reactivities in MS patients was significantly higher than those in normal controls (p = 0.0193). Anti-HSP105 reactivities were found in 3 % (2/66) of normal controls, while 14.5 % (10/69) of patients with MS were positive for anti-HSP105 antibodies. The difference in prevalence of autoantibodies was not statistically significant between OS-MS and C-MS. The HSP105 α-reactive T cell lines derived from CD4^+CD45RO^+ T cells were established from 3 of 9 MS patients but not normal controls. Our findings indicate HSP105 to be a novel candidate autoantigen in MS. In future, we are going to analyze the other candidate autoantigens (NSD1, KIAA1640, KIAA0610). Less

当前假设多发性硬化症（MS）是由自身反应性T细胞针对髓磷脂抗原介导的。在日语中，MS经常和有选择地影响视神经和脊髓。我们先前已经报道说，MS（OS-MS）的视脊髓形式已被证明与临床，免疫学和免疫生成的MS（C-MS）的常规形式不同。最近，已经证明少突胶质细胞具有复数起源，根据其起源而显示出明显的分布差异。我们假设在视神经和脊髓中存在的特定自身抗原可能与OS-MS病变的发展有关。为了鉴定OS-MS中的自身抗原，我们利用了人脊髓cDNA表达文库的免疫统计，并使用患有OS-MS患者获得的血清样品进行了血清样品。从人脊髓制备cDNA表达文库，并用从八名OS-…更多MS患者获得的血清免疫斑块5 x 10^5至10 x 10^5。通过对血清的反应性鉴定了11个阳性免疫反应性克隆，并分析了它们的序列。序列的比较表明，这些克隆代表来自5个不同基因的cDNA（HSP105，Rabaptin-5，NSD1，KIAA1640，KIAA0640）。我们使用ELISA在血清和CSF中生产了GST-Rabaptin-5融合蛋白和HSP105蛋白，并评估了IgG抗Rabaptin-5和抗HSP105自身抗体的滴度。我们发现IgG抗Rabaptin-5反激活率没有显着升高，但是MS患者的IgG抗HSP105反激活率显着高于正常对照组中的IgG抗HSP105反应率（P = 0.0193）。在3％（2/66）的正常对照中发现了抗HSP105反应率，而14.5％（10/69）的MS患者对抗HSP105抗体呈阳性。在OS-MS和C-MS之间，自身抗体的患病率差异在统计学上没有统计学意义。从9个MS患者中的3例中建立了源自CD4^+ CD45RO^+ T细胞的Hsp105α反应T细胞系，但不是正常对照。我们的发现表明HSP105是MS中的新型候选自动抗原。将来，我们将分析其他候选自动抗原（NSD1，KIAA1640，KIAA0610）。较少的