Gene expression profiling in multiple sclerosis brain

Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system and the leading cause of non-traumatic neurological disability in young adults in the United States and Europe. The clinical disease course is variable and starts with reversible episodes of neurological disability in the third or fourth decade of life. Microarray-based comparative gene profiling provides a snapshot of genes underlying a particular condition. Several large scale microarray studies have been conducted using brain tissue from MS patients. In this review, we summarize existing data from different gene expression profiling studies and how they relate to understand the pathogenesis of MS.     

Microarray analysis identifies interferon beta-regulated genes in multiple sclerosis

The molecular mechanisms for the interferon beta (IFNbeta) treatment of multiple sclerosis (MS) remain to be characterized. Using cDNA microarray technology, we have compared the gene expression profile of T and non-T cells derived from relapsing-remitting MS before and after treatment with IFNbeta-1b. IFNbeta treatment significantly altered expression of 21 genes out of 1263 at 3 and 6 months after treatment. These genes included nine with IFN-responsive promoter elements. Whereas there was no change in Th1 or Th2 marker genes, some of the changes were unexpected but coincided with the beneficial effect of IFNbeta in MS.     

精神分裂症患者外周血白细胞基因差异表达的基因芯片研究

目的 研究健康人及精神分裂症患者外周血白细胞基因表达谱，分析精神分裂症患者基因表达水平的变化。方法 取6例精神分裂症患者和6名健康人的外周血白细胞，提取总。RNA，在反转录cDNA时，分别用Cy3、cy5两种激发波长荧光标记，获得两组样本cDNA探针。将两组探针混合后与含8464条人类基因表达谱芯片杂交，扫描、分析杂交结果。结果 当Cy5与Cy3比值大于2或小于0．5时，在6对样本7次试验中有13条基因在精神分裂症患者中均下调表达。当Cy5与Cy3比值大于1．5或小于0．7时，在6对样本7次试验共同出现差异的基因有31条，其中2条基因在精神分裂症患者中上调表达，29条下调表达。结论 精神分裂症患者的一些基因表达水平与正常人存在差异，这些基因主要与免疫、神经细胞发育、神经传递、髓鞘形成有关。     

多发性硬化免疫相关基因表达谱的基因芯片研究

目的：免疫相关基因芯片寻找多发性硬化表达差异的免疫相关基因，方法：将484条免疫相关基因的double(双点）cDNA产物按微矩阵排列点样于玻片上，例1患者及其对照为第1组，例2患者及其对照为第2组，例3患者及其对照为第3组。取各组外周血，分离淋巴细胞，提取总RNA，用RT－PCR逆转录成cDNA。标记cDNA探针，分别用cy-3-dUTP,cy5-dUTP标记正常人和多发性硬化患者cDNA。将基因芯片和杂交探针变性后杂交。洗干。用Scan Array4000扫描芯片，GenePixPro3.0软件分析cy3,cy5两种荧光信号的强度和比值。结果：1组中筛选出差异表达的基因22项，2组中出现差异表达的基因27项，3线中出现差异表达的基因72项，具有同一GeneID号的一对Double基因，其中1组6对，2组9对，3组30例。结论：试验结果显示正常人与MS患者免疫相关基因的表达有显著差异，不同发病阶段免疫相关基因表达有差异；以细胞免疫相关基因变化为主，体液免疫相关基因亦有异常表达。     

多发性硬化患者外周血单个核细胞转录因子Sp3基因的缺陷表达

目的：研究我国多发性硬化（MS）病人外周血单个核细胞（PBMC）转录因子Sp3基因表达情况及其与临床相关性。方法：采用反转录－聚合酶链反应（RT－PCR）技术，经2对引物扩增，检测了33例MS、30例非免疫 性其他神经系统疾病对照、30例神经系统其他自身免疫性疾病对照和30名健康对照组者PBMC的Sp3基因表达。结果：Sp3基因表达缺如，MS患者组（45％，15／33）明显高于其他3组（依次为16％，5／30，6％，2／30；10％，3／10，P均（＜0．01），可自Sp3表达阴性MS患者的DNA中扩增出相应Sp3片段。Sp3表达阴性MS患者改良的伤残状态量表（EDSS）评分显著高于表达阳性者。脑脊液的IgG24h合成率和血清可溶性白细胞介素－2受全稍高，4例MS患者治疗前病情重和治疗后病情轻时相比，未见其Sp3基因表达有明显变化。结果：汉 MS病人PBMC中有Sp3表达缺陷，Sp3阴性患者残疾程度和免疫功能紊乱程度明显高于阳性者，作 为转录调节因子Sp3的表达缺乏可能与免疫控制的异常启动有关。     

Deficient Fas expression by CD4+ CCR5+ T cells in multiple sclerosis

OBJECTIVE: To evaluate whether T cells expressing CCR5 and CXCR3 from multiple sclerosis (MS) patients are more resistant to apoptosis. METHODS: Expression of CD69, TNF-R1, Fas, FasL, bcl-2, and bax was investigated in 41 MS patients and 12 healthy controls by flow cytometry in CD4+ and CD8+ T cells expressing CCR5 and CXCR3. RESULTS: In MS patients, the percentage of CD69 was increased and Fas expression decreased in CD4+ CCR5+ T cells. INTERPRETATION: The lower Fas expression in activated CD4+ CCR5+ T cells might contribute to disease pathogenesis by prolonging cell survival and favoring their migration into the CNS.     

多发性硬化413例患者的临床表现特点

目的 总结多发性硬化(MS)患者的临床特点。方法 用临床病例分析统计方法，对413例MS患者的发病规律和临床特点进行归纳、分析。结果 MS好发于青壮年，以急性和亚急性起病为主；首发症状以视力障碍(128例，31．O％)最常见；肢体无力(325例)、感觉障碍(246例)、视力障碍(243例)是MS患者最常见的症状，Lhermitte征较多见；发作性症状多见，以痛性痉挛发作和癫疴发作常见；可合并周围神经系统损害；临床定位以脊髓(256例)和视神经(244例)受累最多见。结论本组MS临床特点不同于西方人。     

Serum CD95 of relapsing remitting multiple sclerosis patients protects from CD95-mediated apoptosis

Failure of CD95-mediated apoptosis as a potential negative regulatory mechanism of T cell expansion may be involved in T cell-mediated autoimmune diseases such as multiple sclerosis (MS). Recently, soluble CD95 has been shown to be elevated in MS patients with active disease. Here, we report that the sera of MS patients inhibit CD95 ligand-induced apoptosis of susceptible target cells in a concentration-dependent manner and dependent on the amount of serum CD95 levels. Thus, MS sera contain biologically active inhibitors of T cell apoptosis that may allow for prolonged abnormal immune responses.     

Microarray analysis on archival multiple sclerosis tissue: Pathogenic authenticity outweighs technical obstacles

Probably all neuropathologists know this dilemma: on the one hand, they have extremely precious archival material in their possession, which has been collected over many years from many different laboratories. Typically, this material is extremely well characterized, and often, it contains especially significant tissue specimens from unique cases. On the other hand, they face severe scepticism when they plan to use this archival material for large‐scale gene expression studies by microarray analysis, since previous handling in the absence of RNA protection, prolonged storage at room temperature, and fixation with formaldehyde may dramatically reduce the amount of retrievable RNA. Fortunately, this dilemma can be solved. We give here examples from our own, multiple sclerosis‐centered laboratory and explain why archival tissue might be more authentic for the disease process and might yield more information about the molecular and cellular substrates driving CNS inflammation in MS patients than more recently acquired tissues.     

多发性硬化人类白细胞抗原易感基因研究进展

多发性硬化为一种免疫相关性中枢神经系统脱髓鞘疾病,临床表现呈异质性,好发于青壮年女性,病残率高,给患者家庭及社会造成沉重负担.遗传因素和环境因素共同参与其发病机制,而HLA基因复合体为重要的遗传学决定因素,占易感风险的60%,对其研究是目前多发性硬化遗传学研究的热点和难点.     

Single nucleotide polymorphisms and functional analysis of MxA promoter region in multiple sclerosis

OBJECTIVE: Interferons (IFNs)-inducible myxovirus resistance protein A (MxA) has recently been used as an indirect marker of neutralizing antibody against IFN in patients with multiple sclerosis (MS). On the other hand, MxA inhibits the replication of viruses by means of modifying cellular function, including apoptotic pathway. Our objective is to investigate the genetic and pathological role of MxA in patients with MS. METHODS: We examined SNPs of MxA promoter region in 67 patients with MS. Moreover, to elucidate the functional roles of SNPs, we conducted Luciferase assay with pGL3-basic vector including patient-derived or artificially mutated MxA promoter region. RESULTS: A significantly higher frequency of the haplotype with -88T and -123A, which correlates with over-expression of MxA, was observed in MS. Moreover, we elucidated novel findings showing that nt -88 played a leading part with type I IFNs and that nt -123 played the same role independently without type I IFNs, respectively. CONCLUSION: SNPs on MxA promoter region may play an important role in the pathophysiology of MS and provide a novel strategy for the therapeutic resolutions of MS.     

多发性硬化20例临床分析

目的 :探讨多发性硬化的临床症状及MRI对其诊断的价值和意义。方法 :综合分析 2 0例多发性硬化患者一般资料、MRI、脑脊液检查及治疗转归。结果 :多发性硬化好发于 10～ 4 0岁 ,女性多见 ,起病形式及临床症状多样 ,病程呈复发与缓解 ,MRI对多发性硬化病灶的发现率可达 10 0 %。糖皮质激素治疗有效 ,但随着病程的延长 ,复发次数的增多 ,激素治疗欠佳。结论 :根据临床特点、影像学检查、脑脊液及神经电生理检查可提高多发性硬化的确诊率     

Trauma and multiple sclerosis

Summary An obligatory event in the pathogenesis of the multiple sclerosis plaque appears to be an increase in the permeability of the blood-brain barrier. Neuropathological observations of the brain of persons suffering from concussion after relatively minor head injury, as well as of animals subjected to experimental brain injury, have shown that alterations of the blood-brain barrier constitute a common result of such trauma. It is postulated that the alterations of the blood-brain barrier secondary to trauma of the brain or spinal cord of patients with already established multiple sclerosis may result in an exacerbation or recurrence of a previously symptomatic plaque, in the appearance of symptoms from a silent lesion, or in the formation of a new plaque in such an area of selected vulnerability. In other persons injury to the nervous system may cause the development of multiple sclerosis plaques in the previously damaged areas when the disease has its onset after the trauma. There is no evidence to support the idea that trauma ever causes multiple sclerosis.     

Choreoathetosis in multiple sclerosis

Extrapyramidal symptoms are extremely rare in multiple sclerosis. We examined a patient with a ten year history of multiple sclerosis, who developed choreoathetoid movements of both upper extremities. To our knowledge this is the second case report of this association.     

Western and Asian features of multiple sclerosis in Mexican Mestizos

OBJECTIVES: The objective of this study was to compare the clinical expression of MS in Mexican Mestizos with that of patients of European or Asian descent; as well as to compare the annual frequency of new cases with that observed in the previous decades. PATIENTS AND METHODS: All patients with diagnosis of definite MS seen at the National Institute of Neurology and Neurosurgery of Mexico from January 1993 to December 2003 were studied (n=312). Sociodemographic and clinical characteristics were compared with reports of patients from either Western or Asian origin; the long-term disability score was analyzed according to gender, age of onset of MS and the initial symptom. RESULTS: The clinical expression of MS in Mexican Mestizos shares some characteristics with both, Asian and Western forms of MS indicating that the genetic composition of Mexican Mestizos participates in the clinical expression of the disease. Also, at the prevalence date, the mean age of patients and the duration of the disease were lower in our patients than in MS patients from endemic countries suggesting a true increasing incidence in recent times, rather than only improved case ascertainment. CONCLUSIONS: Clinical expression of MS in Mexican Mestizos shows the coexistence of some features common in European and in Asian cases.     

多发性硬化患者外周血单个核细胞转录因子Sp3基因表达缺失及其与免疫的关系

目的 研究多发性硬化(Ms)患者外周血单个核细胞(PBMC)转录因子Sp3基因的表达情况及其与机体免疫功能的关系.方法 贵州地区发病的汉族MS患者31例及健康对照30名,采用逆转录PCR(RT-PCR)检测其PBMC的Sp3基因表达情况;双抗体夹心ELISA法检测血清可溶性白细胞介素-2受体(sIL-2R)水平.结果 MS组与健康对照组PBMC cDNA扩增Sp3基因阴性率差异有统计学意义[分别为41.9%(12/31)和6.7%(2/30),x2=7.133,P=0.008].MS组血清sIL-2R含量较健康对照组明显升高[(2788.5±1079.8)、(1270.6±489.4)μg/L,t=6.170,P=0.001],其中Sp3基因阴性[(3364.0±1252.3)μg/L]、阳性[(2450.0±827.0)μg/L]表达患者血清sIL-2R含量较健康对照组均明显升高(F=32.059,P<0.05),Spa基因阴性表达患者比阳性表达患者sIL-2R含量明显升高(q=4.213,P<0.05).结论 贵州地区发病的汉族MS患者PBMC中存在Sp3基因表达缺失,提示汉族MS患者Sp3基因表达无明显的地域差异.sIL-2R参与了MS的发病过程,Sp3基因表达缺失后机体发生了更严重的免疫功能障碍.     

Investigation of an inducible nitric oxide synthase gene (NOS2A) polymorphism in a multiple sclerosis population

Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system (CNS) affecting most commonly the Caucasian population. Nitric oxide (NO) is a biological signaling and effector molecule and is especially important during inflammation. Inducible nitric oxide synthase (iNOS) is one of the three enzymes responsible for generating NO. It has been reported that there is an excessive production of NO in MS concordant with an increased expression of iNOS in MS lesions. This study investigated the role of a bi-allelic tetranucleotide polymorphism located in the promoter region of the human iNOS (NOS2A) gene in MS susceptibility. A group of MS patients (n = 101) were genotyped and compared to an age- and sex-matched group of healthy controls (n = 101). The MS group was subdivided into three subtypes, namely relapsing-remitting MS (RR-MS), secondary-progressive MS (SP-MS) and primary-progressive MS (PP-MS). Results of a chi-squared analysis and a Fisher's exact test revealed that allele and genotype distributions between cases and controls were not significantly different for the total population (chi(2) = 3.4, P(genotype) = 0.15; chi(2) = 3.4, P(allele) = 0.082) and for each subtype of MS (P > 0.05). This suggests that there is no direct association of this iNOS gene variant with MS susceptibility.     

Linkage analysis of candidate myelin genes in familial multiple sclerosis

Multiple sclerosis (MS) is an autoimmune demyelinating disease of the central nervous system. A complex genetic etiology is thought to underlie susceptibility to this disease. The present study was designed to analyze whether differences in genes that encode myelin proteins influence susceptibility to MS. We performed linkage analysis of MS to markers in chromosomal regions that include the genes encoding myelin basic protein (MBP), proteolipid protein (PLP), myelin-associated glycoprotein (MAG), oligodendrocyte myelin glycoprotein (OMGP), and myelin oligodendrocyte glycoprotein (MOG) in a well-characterized population of 65 multiplex MS families consisting of 399 total individuals, 169 affected with MS and 102 affected sibpairs. Physical mapping data permitted placement of MAG and PLP genes on the Genethon genetic map; all other genes were mapped on the Genethon genetic map by linkage analysis. For each gene, at least one marker within the gene and/or two tightly linked flanking markers were analyzed. Marker data analysis employed a combination of genetic trait model-dependent (parametric) and model-independent linkage methods. Results indicate that MAG, MBP, OMGP, and PLP genes do not have a significant genetic effect on susceptibility to MS in this population. As MOG resides within the MHC, a potential role of the MOG gene could not be excluded. Received: January 12, 1999 / Published online: August 17, 1999