多发性硬化患者病情演变与血浆和脑脊液基质金属蛋白酶-9水平的相关性

目的观察急性期多发性硬化(MS)患者血浆和脑脊液基质金属蛋白酶-9(MMP-9)水平的动态变化规律及其与临床神经功能障碍的相关性,探讨其对疾病活动性的判定价值。方法收集急性期MS患者、缓解期MS患者、健康对照各30例,神经系统非炎性疾病(NIND)17例,采用酶联免疫吸附试验法(ELISA)检测血浆和脑脊液MMP-9水平,并进行EDSS评分。结果 (1)与急性发作期初期相比,急性期MS患者第1、2周血浆MMP-9水平明显升高(P=0.019;P=0.014),第4周其脑脊液水平明显降低(P=0.014)。(2)急性期MS组血浆MMP-9水平明显高于缓解期组和健康对照组(P0.01)。(3)与NIND组比较,急性期MS组脑脊液MMP-9水平明显升高(P0.01)。(4)在MS急性期血浆与脑脊液MMP-9水平呈正相关(r=0.571,P=0.016),两者与患者同期EDSS评分之间无相关性(P0.05)。结论 (1)MS患者血浆MMP-9水平可成为监测疾病活动的指标。(2)脑脊液MMP-9水平对鉴别MS患者与NIND有一定帮助。(3)在一定程度上急性期MS组血浆MMP-9水平可以代表脑脊液中的水平。     

多发性硬化患者外周血和脑脊液淋巴细胞亚群的观察

用碱性磷酸酶抗酸酶法检查了４６例多发性硬化活动期患者外周血和脑脊液的淋巴细胞亚群。结果显示：活动期ＭＳ者外周血ＣＤ＾＋４，ＣＤ＾＋９细胞较对照组减少，ＣＤ＾＋２５细胞，ＣＤ＾＋４／ＣＤ＾＋８比值较对照组升高。ＣＳＦ中ＣＤ＾４，ＣＤ＾＋２５细胞，ＣＤ＾＋４／ＣＤ＾＋８比值较对照组升高，ＣＤ＾＋８细胞降低，且ＣＳＦ中淋巴亚群均高于自身外周血中的相应细胞。     

趋化因子及其受体与多发性硬化

趋化因子(Chemokine)是一类诱导免疫细胞向特定组织部位浸润和积聚的具有趋化作用的细胞因子。趋化因子对靶细胞的特殊作用是由趋化因子受体决定的。本文将阐述趋化因子及其受体的分类、特性及与多发性硬化(MS)的关系。     

多发性硬化与趋化因子及其受体

趋化因子是一类具有化学趋化活性的细胞因子,在炎症反应中起重要作用。本文对趋化因子及其受体的分类,在炎症细胞移行过程中的作用和与中枢神经系统炎症性脱髓鞘疾病多发性硬化之间的关系进行阐述。     

多发性硬化患者趋化因子MCP-1的表达及意义

目的 测定急性期多发性硬化(MS)患者血清及脑脊液中的趋化因子MCP-1的浓度变化,并对其结果作初步探讨。方法 用ELISA法检测20例急性期MS患者,20例其他神经系统疾患的血清/脑脊液中的趋化因子MCP-1的浓度。结果 急性期MS患者血清及脑脊液中的趋化因子MCP-1的浓度与对照组相比均下降。结论 急性期MS患者血清及脑脊液中的趋化因子MCP-1的浓度下降可能与T细胞的不同反应性有关,趋化因子可能参与MS的发病过程。     

脑脊液中CXCL 13与临床孤立综合征及多发性硬化关系的Meta分析

目的评价脑脊液(CSF)中CXCL 13与临床孤立综合征(CIS)及多发性硬化(MS)的关系。方法计算机检索Pubmed(1966²013)、Embase(19742013)、Embase(1974²013)、Ovid(19932013)、Ovid(1993²013)、Cochrane中心临床对照试验注册数据库(CENTRAL)(2011年第2期)、中国生物医学文献数据库(CBMdisc)(19782013)、Cochrane中心临床对照试验注册数据库(CENTRAL)(2011年第2期)、中国生物医学文献数据库(CBMdisc)(1978²013)、CNKI(19792013)、CNKI(1979²013)、VIP(19892013)、VIP(1989²013)及万方数据库(19782013)及万方数据库(1978²013)等,收集关于CXCL 13与CIS及MS关系的队列研究及病例对照研究。按Cochrane系统评价的方法,由2名研究者独立进行质量评价和资料提取,采用RevMan 5.2软件进行Meta分析。结果共纳入6篇的文献,样本量为1011例,其中病例组(MS及CIS)721例;对照组(神经系统非炎性疾病[NND])290例。与NND及CIS组相比,MS患者CSF中CXCL 13水平高于NND及CIS组(P<0.00001);随访2 y,与未转化的CIS组相比,转化为MS的CIS组CSF中CXCL 13水平高于未转化的CIS组(P=0.009);CSF中CXCL13水平>10 pg/ml的CIS患者,转化为MS的转化率高于<10 pg/ml者(P=0.0001)。结论与NND及CIS组相比,MS的CSF中CXCL 13水平明显升高,对CIS转归为MS有重要的预测价值,提示CSF中CXCL 13的水平与MS的发生发展有关。     

β-干扰素对多发性硬化患者趋化因子mRNA表达的影响

目的　观察干扰素 β 1b(IFNβ 1b)在体外对多发性硬化 (MS)患者趋化因子mRNA表达的影响。方法　取MS患者外周血 ,分离单个核细胞 (MNC) ,以其他非炎性神经系统疾病 (OND)及健康人 (HC)为对照组。MNC在完全培养基中分别与自身抗原髓鞘碱性蛋白 (MBP)、对照抗原AChR及不加抗原组 ,加或不加药物IFNβ 1b共同培养 ,3d后收集细胞 ,涂片 ,用地高辛标记的寡核苷酸探针进行原位杂交 (ISH) ,检测C C趋化因子单核细胞炎性蛋白 1α/ β(MIP 1α/ β)、单核细胞趋化蛋白 1(MCP 1 )和正常T细胞表达及分泌的调节活化因子 (RANTES)mRNA的表达。结果　MBP刺激的MIP 1α及自发产生的MIP 1αmRNA均受到IFN β 1b的抑制 ,但差异无显著意义 (P >0 0 5)。RANTESmRNA的表达受到IFNβ 1b的抑制 ,在MBP诱导下无药物处理时为 30 2± 1 5 0 (细胞数 / 1 0 5,下同 ) ,有药物处理为 1 1 1± 5 3 ,差异有显著意义 (P <0 0 1 ) ;在无抗原诱导下无药物处理时为 1 8 5± 3 3 ,有药物处理为 5 1± 3 2 ,差异亦有显著意义 (P <0 0 1 )。IFNβ 1b在 1 0U/ml浓度下 ,可对MBP刺激的MCP 1mRNA的表达产生抑制作用 (分别为 1 58 4± 1 0 4 3和 63 2± 36 9,差异有显著性意义 ,P <0 0 1 ) ,而对自发产生的MCP 1mRNA作用不明显 ;对MBP刺激     

血清和脑脊液CXCL13水平与临床孤立综合征、多发性硬化和视神经脊髓炎的关系

目的探讨临床孤立综合征(CIS)、复发缓解型多发性硬化(RRMS)、视神经脊髓炎(NMO)患者血清和脑脊液内B淋巴细胞趋化因子-1(BLC-1/CXCL13)的水平与疾病进展、扩展残障状态量表(EDSS)评分及MRI表现的关系。方法选择CIS患者18例、RRMS患者22例、NMO患者21例,以及神经系统非炎性疾病(neurological non-inflammatory disease,NND)患者(作为对照组)17例,采用酶联免疫吸附试验法检测4组患者血清和脑脊液CXCL13水平并进行比较;对4组患者进行发病期EDSS评分及MRI检查,比较EDSS评分≥3.5分和EDSS评分<3.5分患者血清和脑脊液CXCL13水平,分析CXCL13水平与EDSS评分的相关性,比较头颅和脊髓增强扫描阳性与阴性患者血清及脑脊液CXCL13水平;随访18例CIS患者2年,比较脑脊液CXCL13水平>10pg/mL患者与脑脊液CXCL13水平<10pg/mL的患者转化为MS的病例数。结果 CIS组、RRMS组及NMO组与NND组患者相比,血清和脑脊液中CXCL13的水平高(均P<0.01),其中RRMS组患者脑脊液中CXCL13的水平较CIS组和NMO组高(均P<0.01);EDSS评分≥3.5分患者血清和脑脊液CXCL13水平比EDSS评分<3.5分患者高(均P<0.01),4组患者血清和脑脊液中CXCL13水平与患者EDSS评分值呈正相关(r=0.881,P<0.01;r=0.753,P<0.01);行头颅和脊髓MRI增强扫描的48例患者中,有增强病灶者脑脊液中CXCL13水平比无增强病灶者高(P<0.01);脑脊液CXCL13水平>15pg/mL的患者转化为MS的比例(37.5%)与脑脊液CXCL13水平<10pg/mL的患者(10.0%)比较无统计学差异(P>0.05)。结论脑脊液CXCL13水平高的CIS患者可能较早地转化为MS,CXCL13可能是预测CIS转化的标记物。     

多发性硬化患者病情演变与tau蛋白关系的研究

目的 探讨神经元损伤的标志物tau蛋白与多发性硬化患者病情演变的关系.方法 48例多发性硬化患者分为复发-缓解组和继发进展组,MSSS和EDSS量表评定疾病严重程度及致残状况.ELISA法测定患者脑脊液和血清总-tau(total-tau,T-tau)蛋白和磷酸化-tau(phospho-tau,P-tau)蛋白浓度.结果 复发-缓解组患者脑脊液T-tau蛋白浓度明显高于继发进展组(P=0.01).多发性硬化患者脑脊液T-tau蛋白与EDSS评分呈负相关(γ=-0.58,P=0.0006).结论 多发性硬化患者病程中脑脊液总tau蛋白浓度的变化,间接反映脑实质损伤的程度.     

多发性硬化患者脑脊液蛋白质组学研究

目的 探讨多发性硬化(MS)发病的分子机制并寻找其脑脊液(CSF)蛋白质标志物.方法 对MS患者与紧张型头痛患者各6例的CSF蛋白质组学进行比较研究.将浓缩的CSF去除高丰度蛋白后行双向凝胶电泳,对凝胶图谱进行图像分析,探寻差异性蛋白点并经肽质量指纹质谱技术鉴定蛋白质点.结果 MS患者38种蛋白质的表达与对照组比较差异有统计学意义(P<0.05),部分差异性蛋白质经鉴定为白蛋白单体CRA_n、ATP合成酶、普通转录因子IIH、白蛋白单体CRA_j、四连接素、结合珠蛋白、丛生蛋白、富亮氨酸α-2糖蛋白、视黄醇结合蛋白与甲状腺素结合蛋白.结论 MS患者CSF多种蛋白质表达差异,可能存在能量代谢异常、炎症反应和组织修复机制.     

Prostaglandin levels in cerebrospinal fluid from multiple sclerosis patients in remission and relapse

Radioimmunoassay (RIA) techniques have been employed to determine prostaglandin (PG) levels in the cerebrospinal fluid (CSF) from multiple sclerosis (MS) patients in remission and relapse and in subjects with other neurological diseases (OND). PGE and PGF₂α concentrations in spinal fluid from MS patients in relapse were significantly lower than values estimated during remission and in individuals with OND of the central nervous system (CNS). These observations are discussed in relation to the clinical state of patients with demyelinating disease together with a consideration of the concept that disordered immune mechanisms contribute a central role in the pathogenesis of MS.     

Growth hormone and insulin growth factor-I levels in plasma and cerebrospinal fluid of patients with multiple sclerosis

Multiple sclerosis (MS) has several clinically different forms. Whereas the illness progresses slowly in most of the patients, 10% have an aggressively progressive course with fatal outcome without signs of remyelination capability. The process of remyelination depends on numerous interactive factors, including the presence of various growth factors, the most important of which in the adult is insulin growth factor-I (IGF-I). On the other hand, the most powerful postnatal regulator of IGF-I is growth hormone (GH), which also acts as a neuroprotective and an antiapoptotic agent, and has direct influence on myelination. Levels of these growth factors have never been examined in the cerebrospinal fluid (CSF) of patients with MS. The levels of IGF-I and GH were measured in serum and CSF of 46 MS patients and compared with those of 49 patients with no evidence of demyelinating disease. The only positive finding was a deficiency of GH in the CSF of MS patients. The possible implications of those findings in the etiopathogenesis of MS will be discussed.     

Expression of leukemia inhibitory factor in the cerebrospinal fluid of patients with multiple sclerosis

Multiple sclerosis (MS) is an inflammatory demyelinating disease of the central nervous system, characterized by infiltration of immune cells in the central nervous system, localized myelin destruction and loss of oligodendrocytes. Early detection of MS may be possible via blood and cerebrospinal fluid (CSF) tests based on disease pathology. Leukemia inhibitory factor (LIF), a neurotrophic cytokine, has previously been shown to limit autoimmune demyelination and oligodendrocyte loss in a murine model of MS. Given its potential role in neural cell death and survival, in the present study we measured expression of LIF in serum and CSF from patients with relapsing-remitting MS (n = 46) and control subjects (n = 42). We used western blot analysis and enzyme-linked immunosorbent assays (ELISA), to study LIF expression. Western blot analysis revealed that LIF was present in all CSF samples, and densitometric analysis showed that relative expression was significantly higher in CSF from patients with MS than in controls (p < 0.001). ELISA analysis showed that the concentrations of LIF in both the serum (87.5 ± 11.46 ng/mL) and CSF (56 ± 10.72 ng/mL) of MS patients were significantly higher than those in control subjects (52 ± 8.23 ng/mL, 7.8 ± 3.76 ng/mL, respectively; p < 0.0001 for both serum and CSF), despite there being no significant difference in total protein concentration between the two groups (p = 0.52 for serum, p = 0.2 for CSF). Our data suggest that serum and CSF concentrations of LIF may provide additional useful information during the differential diagnosis of MS. Our findings also indicate that LIF could be significantly involved in the pathophysiology of MS.     

T-cell subsets in the cerebrospinal fluid and blood of patients with multiple sclerosis

The absolute numbers and ratios of helper/inducer (T4) and cytotoxic/suppressor (T8) T-cells were determined in cerebrospinal fluid (CSF) and blood of patients with multiple sclerosis (MS) and various other neurologic diseases (OND). In patients with MS, the T4:T8 ratio was higher in both blood and CSF, and the increase was significantly greater in CSF than in blood. These findings were due to an increased proportion of T4-lymphocytes in the CSF and to a decreased proportion of T8-cells in blood. These results indicate the need for additional studies of CSF lymphocytes in patients with MS.     

CXCL10 haplotypes and multiple sclerosis: association and correlation with clinical course

CXCL10 (interferon- γ -inducible protein-10) levels are increased in cerebrospinal fluid of multiple sclerosis (MS) patients with symptomatic attacks of inflammatory demyelination, supporting a role for this molecule in MS pathogenesis. Two hundred and twenty-six patients with MS and 235 controls were genotyped for G  →  C and T  →  C single nucleotide polymorphisms (SNPs) in exon 4 of CXCL10 gene. Haplotypes were tested for association and correlated with clinical variables. The two SNPs studied were in complete linkage disequilibrium. None of the determined haplotypes was associated with MS. However, carriers of the GGTT haplotype (defined as wild type, according to the sequence in National Centre for Biotechnology Information (NCBI) database) had a significantly lower progression index than non-carriers ( P  = 0.016). Furthermore, amongst patients who had an initial relapsing remitting (RR) course of the disease, the time between onset and second episode was significantly longer in GGTT carriers ( P  = 0.021). Considering secondary progressive (SP)–MS patients, the time between the initial RR form and the subsequent worsening to SP was longer in this group ( P  = 0.08). Therefore, the GGTT haplotype of the CXCL10 gene is not a susceptibility factor for the development of MS, but is probably to influence the course of MS, possibly contributing to slow down the progression of the disease.     

Evaluation of interleukin 10 and interferon-α in the cerebrospinal fluid and serum of patients with multiple sclerosis

The activities of IL-10, a cytokine produced by TH2 cells, monocytes and B-cells, and IFN-α, a product of activated macrophages/monocytes, were investigated in the CSF and serum samples of 25 subjects with clinically definite multiple sclerosis (MS), of whom 14 were in the active and nine in the stable phase, and of 15 controls with other noninflammatory neurological diseases (OND). Elevated CSF IL-10 and IFN-α levels were found in MS patients in the stable phase with respect to patients in the active phase (p > 0.001), while no significant differences were observed in the mean serum levels between MS patients (both in the active and stable phase) and controls. Finally, a significant correlation was detected between IL-10 and IFN-α in the CSF of MS patients in remission. This study suggests that IL-10 may downregulate MS progression.     

Myelin basic protein in cerebrospinal fluid of patients with multiple sclerosis and other neurological diseases

Summary Cerebrospinal fluid (CSF) from 105 patients was analyzed by radio-immunoassay for the presence of material cross-reactive with peptide 89–169 of bovine myelin basic protein (BP).In a group of 72 multiple sclerosis patients, 52 showed higher BP content than the control group, i.e. more than 2 ng/ml CSF. Increased BP or BP fragments could be detected in CSF from almost all patients who recently (within 2 weeks) had had an acute episode, or after deterioration in the progressive form of the disease. Fifteen to 30 days after the onset of exacerbation or in a stable period, BP content decreases and in the slowly progressive form was in the range of the control group with one exception.BP content was also elevated in the CSF of patients with other neurological diseases. The presence of BP in the CSF from patients with isolated retrobulbar neuritis is of particular interest. Thus the presence of material cross-reactive with BP fragment 89–169 is not specific for multiple sclerosis, but is a useful parameter in diagnosis and evaluation of MS.

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Zusammenfassung Mit Hilfe eines Radioimmunoassays wurde der Liquor von 105 Patienten auf Substanzen untersucht, die mit dem Fragment 89–169 des bovinen basischen Myelinproteins (BP) kreuzreagieren. Von 72 Patienten mit Multipler Sklerose hatten 52 einen höheren BP-Gehalt im Liquor als die Kontrollgruppe, d.h. mehr als 2 ng/ml. Dies war bei fast all jenen Patienten der Fall, bei denen der Liquor innerhalb von 2 Wochen nach einem akuten Schub bzw. zum Zeitpunkt einer deutlichen Verschlechterung des Zustandes bei der chronisch-progredienten Form gewonnen wurde. Der BP-Gehalt des Liquors nimmt zwischen dem 15. und 30. Tag nach Schubbeginn deutlich ab. In der schleichenden progredienten Form wurde mit einer Ausnahme kein erhöhter BP-Gehalt gefunden.Das basische Myelinprotein ist aber auch im Liquor von Patienten mit anderen neurologischen Erkrankungen zu finden. Hervorzuheben ist das Auftreten von BP im Liquor von Patienten mit Retrobulbär-neuritis.Der Nachweis von mit Bruchstück 89–169 kreuzreagierenden Substanzen im Liquor ist infolgedessen zwar nicht spezifisch für die Multiple Sklerose, ist aber ein wertvoller Parameter in der Diagnose und Einschätzung der MS.

     

Effect of the treatment with methylprednisolone on the cerebrospinal fluid and serum levels of CCL2 and CXCL10 chemokines in patients with active multiple sclerosis

BACKGROUND: Several experimental and human studies suggest that the chemokines CCL2 and CXCL10 may play a role in the pathogenesis of multiple sclerosis (MS). Here, we evaluated the effect of intravenous methylprednisolone (IVMP) therapy on the levels of CCL2 and CXCL10 in the cerebrospinal fluid (CSF) and serum of patients with active MS. METHODS: Serum and CSF samples were obtained from 14 patients with active relapsing-remitting MS (age +/- SD years, 37.0 +/- 8.1; M/F, 6/8) and age- and gender-matched control subjects. All patients were submitted to IVMP treatment (500 mg daily for 5 days). Blood and CSF sampling were performed at admission, i.e. before treatment (day 0), at the end of the treatment (day 6) and 30 days after treatment (day 30). The clinical status of MS patients was also assessed. CCL2 and CXCL10 were measured by enzyme-linked immunosorbent assay. RESULTS: Multiple sclerosis patients had lower CCL2 and higher CXCL10 in CSF when compared with control subjetcs. After treatment with methylprednisolone, MS patients showed clinical improvement and the CSF concentrations of CCL2 and CXCL10 modified toward normal values. CONCLUSIONS: The clinical improvement of active MS following the treatment with methylprednisolone was associated with the modification of CSF levels of CCL2 and CXCL10, suggesting that these chemokines may be useful markers of response to treatment and relapses in MS patients.