β-干扰素治疗继发进展型多发性硬化临床疗效的荟萃分析

目的系统评价β-干扰素(interferon-β,IFN-β)维持治疗继发进展型多发性硬化的临床疗效。方法采用Cochrane系统评价方法,通过电子检索MEDLINE、EMbas、CBMdisc、Cochrane LibraryI、SI和美国临床试验登记中心等的数据库文献。评价纳入文献的方法学质量后,采用RevMan 4.2.10软件对提取的数据进行分析。结果共纳入4个RCTs研究。IFN-β治疗组与对照组比较,EDSS评分显示的2年疾病进展率差异有统计学意义(P=0.02<0.05,OR=0.82,95%CI=0.70~0.97),EDSS评分显示的3年疾病进展率差异无统计学意义(P=0.29>0.05,OR=0.85,95%CI=0.62~1.15),平均年复发率差异有统计学意义(P<0.00001,OR=0.48,95%CI=0.41~0.57)。结论IFN-β维持治疗继发进展型多发性硬化能降低EDSS评分显示的2年疾病进展率,不能降低EDSS评分显示的3年疾病进展率,可明显降低平均年复发率。     

多发性硬化患者外周血CD4~+CD25~+调节性T细胞FOXP3 mRNA的表达及临床意义

目的探讨多发性硬化（MS）患者外周血CD4^＋CD25^＋调节性T细胞数量以及叉头样转录因子（FOXP3）表达水平及其临床意义。方法（1）按Poser诊断标准结合头颅MRI增强扫描,排除合并其他神经系统疾病和免疫系统疾病,选择临床确诊和临床可能的MS患者53例（男15例,女38例）。所有患者近期均未行特殊免疫治疗。以统一标准进行登记临床评价、扩展的残癌状态评分;以健康体检者作为对照组,共51例（男14例,女37例）;（2）流式细胞仪检测外周血CD4^＋CD25^＋调节性T细胞数量的变化;（3）免疫磁珠分离出CD4^＋CD25^＋T细胞,RT-PCR法检测CD4^＋CD25^＋T细胞FOXP3 mRNA的表达,并进行半定量分析。结果（1）MS患者外周血中CD4^＋CD25high T细胞数量与正常组比较没有明显变化（P〉0.05）,但在活动期低于正常组,且差异有统计学意义（P〈0.05）;（2）同一个体在疾病活动期外周血中CD4^＋CD25 high T细胞数量较非活动期减少（P〈0.05）;（3）MS患者外周血中CD4^＋CD25^＋T细胞的FOXP3 mRNA表达降低（P〈0.05）,在活动期降低更明显（P〈0.01）。结论（1）就总体而言MS患者外周血CD4^＋CD25 high T细胞数量变化不明显,但CD4^＋CD25 high T细胞抑制活性降低,FOXP3 mRNA表达减少且与MS疾病活动性有关;（2）就个体而言MS患者外周血中CD4^＋CD25 high T细胞数量与疾病的活动性有关。     

多发性硬化患者外周血CD4+CD25+ T细胞变化及其机制探讨

目的探讨多发性硬化(MS)患者外周血CD4 CD25 T细胞数量及叉头样转录因子(FOXP3)表达水平与MS病情的关系。方法选择温州地区MS患者44例(男12例、女32例),均按Poser诊断标准诊断,结合头颅MRI增强扫描排除合并其他神经系统和免疫系统疾病,并统一行EDSS评分;对照组43例(男13例、女30例)为健康查体者。具体方法:流式细胞仪检测外周血CD4 CD25 T细胞数量;免疫磁珠法分离CD4 CD25 T细胞;RT-PCR法检测CD4 CD25 T细胞FOXP3 mRNA表达并进行半定量分析。结果MS患者外周血中CD4 CD25 调节性T细胞数量与对照组比较无明显变化(P>0.05);活化的效应性T细胞数量增加(P<0.05)且活动期增加更为显著(P<0.01)。同一个体疾病活动期外周血CD4 CD25 调节性T细胞数量较非活动期减少(P<0.05)。MS患者外周血中CD4 CD25 T细胞的FOXP3 mRNA表达降低(P<0.05),且活动期降低更明显(P<0.01)。结论此组MS患者外周血CD4 CD25 调节性T细胞抑制活性降低,FOXP3 mRNA表达减少,活化的效应性T细胞数量增加,且与MS疾病活动性有关。     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

实验性变态反应性脑脊髓炎AQP-4、CD4~+CD25~+调节性T细胞研究

目的:以髓鞘少突胶质细胞糖蛋白细胞外免疫球蛋白样结构域(MOGIgd)为抗原免疫C57BL/6小鼠,建立EAE模型,并检测AQP-4、CD4+CD25+T细胞的表达水平。方法:①采用分子克隆技术诱导表达MOGIgd-TrxA融合蛋白,纯化后以此融合蛋白为抗原于侧腹壁皮下多点注射免疫C57BL/6小鼠作为实验组(MOG组);同时以硫氧还蛋白(TrxA)、豚鼠脊髓匀浆(GPSCH)免疫小鼠分别作为阴性、阳性组。评估动物的临床神经功能、组织病理情况,评价模型的质量。②免疫组化法、荧光定量PCR技术测定AQP-4表达水平;经流式细胞仪(FACS)检测小鼠脾脏中CD4+CD25+调节性T细胞。结果:①MOG组及GP-SCH组小鼠均呈慢性非缓解型病程,两组在发病时间、达峰时间、临床神经功能评分等方面差异无统计学意义(P0.05)。TrxA组无一动物发病。模型组(包括MOG组和GPSCH组)动物HE染色可见血管周围炎性细胞浸润,胶质结节形成,髓鞘染色可见斑片状髓鞘脱失,大脑、小脑、脑干和脊髓均有不同程度受累。免疫组化定性和半定量分析显示MOG组和GPSCH组病灶周围炎症浸润处AQP-4表达与TrxA组相比明显增高(P0.05);②MOG组CD4+CD25+T细胞占CD4+T细胞比例为(4.71±1.61)%,GPSCH组为(1.44±0.65)%,均明显低于TrxA组(P0.01),且MOG组和GPSCH组之间的差异亦具有统计学意义(P0.01)。结论:用MOGIgd免疫C57BL/6小鼠诱导EAE模型稳定,发病率高,为今后进一步研究MS的免疫发病机制和采取有效治疗措施打下基础。EAE模型动物AQP-4的表达量与炎症浸润的严重程度相关。     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

目的 探讨多发性硬化(MS)患者外周血CD8+CD28-T细胞数量的变化及临床意义.方法 采用前瞻性病例对照研究,收集自2005年10月至2008年8月间在温州医学院附属第一医院神经专科病门诊或病房治疗的处于急性活动期的复发缓解型MS患者51例,均符合2005年修改的Mc-Donald诊断标准[2].所有住院患者均给予甲基强的松龙1.0 g/d,连续使用5 d后改为强的松60mg/d,12 d后减量,总疗程不超过6周,对14例MS患者进行治疗前后的动态观察.以20例健康体检者为对照组(NC组),MS组和NC组在年龄、性别构成上差异无统计学意义.采用流式细胞技术检测外周血CD8+CD28-,CD8+CD28+,CD8+及CD4+CD8-T细胞的百分比.两组间均数比较采用独立样本t检验,治疗前后比较采用配对样本t检验,相关性分析采用Pearson相关检验.结果 急性活动期MS组CD8+CD28-T细胞的百分比为(18.48±9.89)%,低于NC组的(24.48±4.86)%(P<0.01),但CD8+CD28+T细胞百分比为(12.23±4.31)%,高于NC组的(8.55±3.49)%(P<0.01),CD8+T细胞的百分比两组比较差异无统计学意义(P>0.05);MS组CD8+CD28-与CD4+CD8-T细胞的百分比呈负相关性(r=0.488,P<0.01);MS患者激素治疗后CD8+CD28-和CD8+CD28+T细胞卣分比与治疗前相比差异无统计学意义(均P>0.05),且治疗后CD8+CD28-T细胞百分比(16.22±4.25)%低于NC组(P<0.01),CD++CD28+T细胞百分比(13.04±4.23)%高于NC组(P<0.01).结论 CD8+CD28-T细胞数日的减少参与了MS的病理过程,并可能通过调节CD4+T细胞起作用;MS急性期激素治疗并不能恢复CD8+CD28-T细胞的数目,激素可能是通过其他途径发生作用.     

C57BL/6小鼠EAE模型的建立及Foxp3、CD4~+CD25~+调节性T细胞检测

目的 以髓鞘少突胶质细胞糖蛋白细胞外免疫球蛋白样结构域(MOG~(Igd))为抗原免疫C57BL/6小鼠,建立实验性自身免疫性脑脊髓炎(EAE)模型,并检测CD4~+CD25~+T细胞、Foxp3 mRNA等指标的表达.方法 采用分子克隆技术诱导表达MOG~(Igd)-TrxA融合蛋白,纯化后以此融合蛋白为抗原于侧腹壁皮下多点注射免疫C57BL/6小鼠作为实验组(MOG组);同时以硫氧还蛋白(TrxA)、豚鼠脊髓匀浆(GPSCH)、生理盐水免疫小鼠分别作为阴性、阳性及正常对照组.双盲法连续早晚观察30 d,通过评估动物的临床神经功能、组织病理(HE染色和Kluver-Barrera法髓鞘染色、免疫组化法)情况,评价模型的质量.流式细胞仪(FACS)检测小鼠脾脏中CD4~+CD25~+调节性T细胞;real-time PCR检测Foxp3 mRNA的相对表达水平,并分析两者相关性.结果 MOG组及GPSCH组小鼠均呈慢性非缓解型病程,两组在发病时间、达峰时间、临床神经功能评分等方面差异均无统计学意义(P>0.05).TrxA组及正常对照组无一动物发病.模型组(包括MOG组和GPSCH组)动物HE染色可见血管周围炎性细胞浸润,胶质结节形成,髓鞘染色可见斑片状髓鞘脱失,大脑、小脑、脑干和脊髓均有不同程度受累.免疫组化定性和半定量分析显示MOG组和GPSCH组病灶周围炎症浸润处AQP-4表达与TrxA组相比明显增高(P<0.05);MOG组CD4~+CD25~+T细胞占CD4~+T细胞比例为(4.71±1.61)%,GPSCH组为(1.44±0.65)%,均明显低于TrxA组和正常对照组(P<0.01),且MOG组和GPSCH组之间的差异亦具有统计学意义(P<0.01).MOG组标准化Foxp3 mRNA的表达量为2.26±1.97,GPSCH组为1.44±1.20,均低于TrxA组的8.58±3.34(P<0.01);但MOG组和GPSCH组间的差异无统计学意义(P>0.05).相关性分析显示,模型组小鼠CD4~+ CD25~+调节性T细胞与Foxp3 mRNA表达水平间存在相关性(r=0.849,P<0.05).结论 用MOG~(Igd)免疫C57BL/6小鼠诱导EAE模型稳定,发病率高,为今后进一步研究多发性硬化的免疫发病机制和采取有效治疗措施打下基础.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.     

多发性硬化患者外周血CD8+CD28-调节性T细胞的变化

Objective To investigate the levels of peripheral blood CD8+ CD28- regulatory lymphocytes and their clinical values in the patients with multiple sclerosis (MS). Method From October 2005 to August 2008, 51 patients with active rehpsing-remitting MS were enrrolled from Department of Neurology of the First Affil-iated Hospital of Wenzliou Medical College. The diagnostic criteria for MS were the 2005 revisions to the "McDon-ald criteria". All the admitted patients received 1 g of methylprednisoione per day intravenously for 5 days, fol-lowed by 60 mg prednisone per day orally for 12 days,and tapered in 6 weeks. Fourteen patients were reevaluated after corticosteroid therapy. Twenty healthy individuals ,as normal controls,matched for age and sex with the MS patients were also enrolled in this study. The percentages of peripheral blood T cells (CD8+ CD28-, CD8+CD28+, CD8+, CD4+ CD8-) were measured by flow cytometric analysis. Parametric statistical analysis were per-formed using standard methods, and linear regression analysis was conducted using Pearson correlation test. Re-sults (1)Compared with controls,the patients with active MS had significantly lower percentage of CD8+ CD28-T cells [(18.48±9.89)% vs. (24.48±4.86)%, P <0.01], and higher percentage of CD8+ CD28+ T cells [(12.23±4.31) % vs. (8.55±3.49) %, P <0.01]. (2)The percentage of CD8+ CD28- T cells was negative-ly correlated with that of CD4+ CDS- T cells (r = -0.488, P < 0.01). (3) After corticosteroid therapy, the per-eentage of peripheral blood CD8+ CD28- / CD8+ CD28+ T cells didn' t significantly decrease or increase in 14 ac-tive MS patients (P > 0.05). Conclusions The decrease of peripheral blood CD8+ CD28- regulatory T cells might be associated with the pathogenesis of MS, and CD8+ CD28- regulatory T cells perhaps played their roles through CD4+ T cells. Corticosteroid therapy could not reverse the levels of CD8+ CD28- T cells.