Wistar大鼠实验性变态反应性脑脊髓炎的模型建立

目的:建立Wistar大鼠多病程实验性变态反应性脑脊髓炎(EAE)的动物模型,并进行病理学研究,为多发性硬化(MS)的研究提供实验依据。方法:以豚鼠全脊髓匀浆(GPSCH)为抗原免疫Wistar大鼠建立EAE的动物模型,并在光镜下观察不同发病类型EAE的病理改变。结果:根据病理和临床表现可将Wistar大鼠EAE模型分为5种发病形式:急性型、缓解-复发型、持续进展型、良性型和隐匿型。光镜下可见不同发病时期的EAE的病理改变有所不同,但都以血管"袖套"状改变、脑室周围及白质脱髓鞘改变为主,伴有神经元肿胀变性。结论:首次建立了Wistar大鼠多病程EAE,是研究多发性硬化的理想动物模型。     

实验性变态反应性脑脊髓炎的病理研究

目的　研究实验性变态反应性脑脊髓炎 (EAE)的病理变化。方法　利用光镜、电镜观察豚鼠 EAE模型的组织学改变。结果　光镜下可见脑实质及珠网膜下腔小血管充血 ,血管周围炎细胞浸润 ,呈“袖套”状改变 ,侧脑室旁白质及脊髓后联合、侧角出现髓鞘肿胀、崩解或消失。电镜下可见血管周围明显脱髓鞘 ,髓鞘呈松散状 ,部分区域斑块状脱落 ,明暗相间结构消失 ,轴突细胞器消失 ,神经元细胞尼 (氏 )小体消失 ,内质网扩张脱颗粒。结论　 EAE的病理改变主要为血管周围炎性浸润及白质脱髓鞘。     

雌激素对实验性变态反应性脑脊髓炎大鼠发病影响的研究

目的探讨雌激素对实验性变态反应性脑脊髓炎(EAE)的影响。方法将30只大鼠随机分为EAE组及大、小剂量雌激素干预组,每组10只,制作EAE模型,大、小剂量雌激素干预组分别给予皮下注射苯甲酸雌二醇1 mg/kg/d、250μg/kg/d,连续10 d,观察各组的发病情况并采用HE染色观察脑和脊髓组织病理变化。结果大、小剂量雌激素干预组的临床症状均较EAE组轻,表现为发病率减少、潜伏期延长、进展期缩短、高峰期神经功能损害较轻。病理切片提示雌激素干预组脑和脊髓炎症细胞浸润明显减少,其中以大剂量组更明显。结论雌激素对多发性硬化动物模型EAE具有保护作用,且与剂量相关。     

山茱萸总苷对大鼠实验性变态反应性脑脊髓炎的初步观察

目的：观察山茱萸总苷（TGCO）对大鼠实验性变态反应性脑脊髓炎（EAE）的影响。方法：Wistar大鼠24只随机分为3组：正常对照组、EAE组和TGCO组。采用动物行为学、常规苏木精-伊红染色和LFB髓鞘染色观察大鼠的发病情况与中枢神经系统（CNS）的病理变化。结果：EAE组7／8只大鼠出现典型的EAE行为学改变、CNS炎性细胞浸润和髓鞘脱失。TGCO组有3,8只大鼠出现EAE行为学改变,评分最高为29,潜伏期延长,CNS内炎性细胞浸润和髓鞘脱失明显减轻。对照组未见动物行为学和CNS病理学改变。结论：TGCO能减轻EAE大鼠的临床症状、延长潜伏期和降低发病率。     

实验性变态反应性脑脊髓炎模型研究进展

实验性变态反应性脑脊髓炎是人类多发性硬化的理想动物模型,在临床神经免疫学研究中具有重要意义。本文主要介绍了实验性变态反应性脑脊髓炎模型制备的有关问题,包括敏感动物品系、性别、年龄的选择,抗原的种类及选择,免疫方法,模型的制备及病情评分等。     

实验性变态反应性脑脊髓炎与脑屏障变化

脑屏障改变是实验性变态反应性脑脊髓炎 (EAE)较早期的病理改变 ,它促使免疫细胞及免疫信息分子大量入脑 ,从而引起脑组织的免疫炎性反应。本文对脑屏障概念 ,EAE时脑屏障变化情况及其可能的机理作一综述。     

T淋巴细胞增殖在实验性变态反应性脑脊髓炎发病中的作用

目的　探讨T淋巴细胞增殖在实验性变态反应性脑脊髓炎 (EAE)发病中的作用。方法　观察Lewis大鼠主动EAE临床症状和T淋巴细胞增殖。结果　EAE临床发病过程中 ,T淋巴细胞增殖旺盛。结论　EAE时 ,活化的T淋巴细胞可以通过BBB进入CNS ,浸入CNS的淋巴细胞能产生破坏性的细胞因子TNF α及某些酶类 ,推测T淋巴细胞增殖活跃 ,在导致EAE发病过程中起着至关重要的作用。     

实验性变态反应性脑脊髓炎IL-10与IL-18的研究

目的研究IL-10与IL-18在实验性变态反应性脑髓炎(EAE)免疫学发病机制中的变化与作用。方法应用豚鼠诱导Wistar大鼠EAE模型,以豚鼠髓鞘碱性蛋白(MBP)与弗氏完全佐剂(CFA)免疫Wistar大鼠,在第11、18、25d处死大鼠,并采用酶联免疫吸附试验(ELISA)测定IL-10与IL-18水平。结果EAE组的IL-10水平在疾病缓解期升高,IL-18的水平随着疾病的进展逐渐升高,在缓解期有所下降,但均显著高于对照组。结论IL-10与IL-18在EAE的免疫学发病机制中具有重要的作用。     

多病程实验性自身免疫性脑脊髓炎大鼠模型的信号转导通路基因芯片研究

目的用信号转导通路发现者基因芯片寻找多病程实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)大鼠不同病程的基因表达差异,从而寻找与其发病机制相关的基因。方法采用豚鼠全脊髓匀浆诱导Wistar大鼠建立EAE模型,分别取正常大鼠、发病期大鼠、缓解期大鼠、复发期大鼠的脊髓腰膨大组织,于-70℃冰箱保存。提取样品RNA,逆转录反应生成cDNA,标记探针,探针与芯片杂交,加入链亲和素耦联的碱性磷酸酶(AP)及化学发光底物反应。图像采集使用GEArray Expression Analysis Suite软件进行完整的芯片数据分析。结果发病期、缓解期、复发期EAE大鼠分别与正常大鼠相比,有表达差异的基因数据为19、23、10项;而有10项基因在比较中随病程变化呈现出发病期表达上调,缓解期表达下调,复发期表达再次上调的趋势。结论实验结果显示EAE大鼠不同发病阶段基因表达有差异,且随病程发展呈一定的趋势变化,这些差异基因与多条信号转导通路有关。     

实验性变态反应性脑脊髓炎相关细胞因子

实验性变态反应性脑脊髓炎(EAE)是一种以细胞免疫为主,以中枢神经系统白质损害为特征的自身兔疫病。在其发病机理及病情演化中,由细胞因子构成的免疫网络起着重要的调节作用,其中一类是由单核巨噬细胞及Th1细胞分泌的致炎细胞因子能够促使EAE的发生或加重病情,而由Th2细胞分泌的免疫抑制性细胞因子又可抑制或逆转EAE的病情。本文收集近年来对EAE相关细胞因子的研究进行综述,以求反映人类多发性硬化疾病的免疫病理。同时还对近来在细胞因子的检测方法及细胞因子在EAE模型的预防和治疗中作用的研究新进展进行了介绍。     

MMP-9和MMP-2在多病程大鼠EAE发病过程中的变化

目的:研究MMP-2和MMP-9在多病程EAE不同类型间的变化,探讨MMPs在MS发病过程中的作用机制。方法:建立多病程大鼠EAE模型,以免疫组化的方法检测MMP-2、MMP-9在不同类型EAE中的表达及分布。结果:MMP-2、-9在EAE中的表达是一致的,不同类型EAE表达MMP-2、-9是不同的。1急性型EAE:炎细胞、血管内皮细胞及细胞外基质、脑膜细胞均呈阳性表达。2缓解-复发型EAE:活动性病灶呈阳性表达,非活动性病灶呈阴性表达。3持续进展型EAE:与缓解复发型EAE表达类似,但呈阳性表达的血管内皮细胞数目多于缓解-复发型。4良性型EAE:只有部分胶质细胞呈阳性表达。5隐匿型EAE-多数病灶呈轻度表达。结论:1 MMP-2、MMP-9在EAE中均有表达。2不同类型EAE中MMP-2、MMP-9的表达是不同的,与病理改变和疾病进展是一致和同步的。     

实验性变态反应性脑脊髓炎发病初期血脑屏障的变化及基质金属蛋白酶的作用研究

目的:探讨多发性硬化的动物模型-实验性变态反应性脑脊髓炎(EAE)发病初期血脑屏障(BBB)的变化及基质金属蛋白酶(MMPs)的作用机制。方法:建立Wistar大鼠EAE动物模型,取发病初期未出现临床症状的EAE大鼠,取脊髓组织通过共聚焦显微镜观察伊文思兰在BBB中的通透性,并用免疫组化的方法测定免疫球蛋白(IgG)的沉积和基质金属蛋白酶(MMPs)的分布。探讨发病初期BBB的变化。结果:CM观察发现,未发病的EAE大鼠沿脊膜下有桔红色荧光分布,同样,EAE大鼠脊髓切片可见有IgG渗出,表现为神经纤维网和神经胶质细胞胞浆呈棕褐色染色,MMP9,-2在脊膜和血管内皮细胞内呈阳性表达,而对照组均呈阴性表达。结论:BBB在EAE发病初期已经受损,MMPs是BBB破坏的重要因素,有降解血管基底膜的作用,可促使炎细胞侵入中枢神经系统。     

Astrogliosis in EAE spinal cord: derivation from radial glia, and relationships to oligodendroglia

A prominent feature of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE) is the accumulation of enlarged, multipolar glial fibrillary acidic protein (GFAP) and brain lipid binding protein (BLBP) immunoreactive astroglia within and at the margins of the inflammatory demyelinative lesions. Whether this astrogliosis is due to both astroglial hyperplasia and hypertrophy or solely to astroglial hypertrophy is controversial. We now report that coincident with the first appearance of inflammation and clinical deficits in mice with myelin oligodendrocyte glycoprotein peptide (MOG peptide)-induced EAE, the radially oriented, bipolar, GFAP, and BLBP positive cells (adult radial glia) present in normal spinal cord white matter undergo mitosis and phenotypic transformation to hypertrophic astroglia. To facilitate visualization of relationships between these hypertrophic astroglia and dying and regenerating oligodendroglia, we used mice that express enhanced green fluorescent protein (EGFP) in cells of the oligodendroglial lineage. During the first week after onset of illness, markedly swollen EGFP+ cells without processes were seen within lesions, whereas EGFP+ cells that expressed immunoreactive cleaved caspase-3 were uncommon. These observations support the hypothesis that necrosis contributes to oligodendroglial loss early in the course of EAE. Later in the illness, EGFP+ cells accumulated amongst hypertrophic astroglia at the margins of the lesions, while the lesions themselves remained depleted of oligodendroglia, suggesting that migration of oligodendroglial lineage cells into the lesions was retarded by the intense perilesional gliosis.     

Epitope spreading occurs in active but not passive EAE induced by myelin basic protein

Using experimental allergic encephalomyelitis, EAE, as a model for the study of autoimmune demyelinating disease in the CNS, previous studies have indicated that spread may occur with respect to the specificity of T cell responses during disease. This phenomenon, known as epitope spreading, is central to therapeutic strategies in multiple sclerosis (MS). However, in EAE, the clinical course, neuropathology and immunopathogenesis vary depending upon host factors and the method of disease induction. Since passive EAE in SJL/J mice resembles MS clinically and neuropathologically, this model was chosen to study the immune phenomenon of epitope spreading. T cells specific for whole 18.5 kDa MBP were used to initiate disease since MBP or one of its naturally occurring cleavage fragments may initiate a more physiological immune response than one generated to an artificially designed synthetic peptide. While a progressive increase in T cell responsiveness specific for the immunodominant MBP 87–106 region was observed during disease, there was no evidence of either intermolecular epitope spreading to the immunodominant region of proteolipid protein (PLP) 139–151 or of intramolecular epitope spreading to the exon 2 encoded region of MBP, which is spliced out of 18.5 kDa MBP. In addition there was no shift in immunodominance toward the subdominant MBP 16–35 region during disease. In contrast, during active EAE induced by MBP, epitope spreading to the immunodominant epitope of PLP, 139–151, was observed. These data demonstrate that immune responses generated during passive versus active EAE may differ, and suggest that significant epitope spreading does not occur in chronic relapsing demyelinating disease initiated with T cells specific for whole MBP in the absence of exogenous antigen, complete Freund's adjuvant and pertussis. Implications of these findings with regard to epitope spreading in MS are discussed.     

Ribavirin reduces clinical signs and pathological changes of experimental autoimmune encephalomyelitis in Dark Agouti rats

The effect of ribavirin on development of experimental autoimmune encephalomyelitis (EAE) was investigated. The disease was induced in genetically susceptible Dark Agouti rats with syngeneic spinal cord homogenate in complete Freund's adjuvant (SCH-CFA). Depending on the amount of mycobacteria in CFA, the animals developed either moderate or severe EAE. Ribavirin (1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide) was applied i.p. at a daily dosage of 30 mg/kg in two treatment protocols: from the start of immunization (preventive treatment) or from the onset of the first EAE signs after the induction (therapeutic treatment). Signs of EAE began between 7 and 9 days after induction and peaked at days 11-13. In moderate EAE (mean maximal severity score 3.33 +/- 0.21), the recovery was completed by days 23-26, whereas, in severe EAE (mean maximal severity score 4.5 +/- 0.23), obvious recovery was not detected. Preventive ribavirin treatment significantly decreased clinical signs after both moderate (score 1.75 +/- 0.25, P < 0.05) and severe (score 3.62 +/- 0.31, P < 0.015) immunization. Also, disease manifestations were reduced by therapeutic treatment of ribavirin (mean maximal severity score 2.5 +/- 0.2 vs. 3.33 +/- 0.21 in controls, P < 0.005) but less so in comparison with preventive treatment. Analysis of the effects of ribavirin on histopathologic changes in the spinal cord tissue revealed a reduction of mononuclear cell infiltrates, composed of T cells and macrophages/microglia, and the absence of demyelination, which were pronounced in control EAE animals. Beneficial effects of preventive and therapeutic treatment with ribavirin on development of EAE suggest this nucleoside analogue as a useful candidate for therapy in multiple sclerosis.     

Myelin basic protein specific T cell lines and clones derived from the CNS of rats with EAE only recognize encephalitogenic epitopes.

The epitope specificities of myelin basic protein (BP) specific T cell lines derived from the spinal cords (SC) and lymph nodes (LN) of rats with experimental autoimmune encephalomyelitis (EAE) were compared. To induce EAE, Lewis rats were immunized with guinea pig (GP)-BP and complete Freund's adjuvant. Mononuclear cells from the SC and LN of these animals proliferated in response to BP and the purified protein derivative (PPD) of mycobacterium. After initially being cultured in growth medium, SC mononuclear cells had an enhanced response to BP and lost their response to PPD. LN cells cultured in identical conditions lost their response to both BP and PPD. LN-derived BP specific cell lines recognized only two epitopes of GP-BP: an encephalitogenic epitope in residues 72-89 and a non-encephalitogenic epitope in residues 43-68. SC-derived BP specific cell lines and clones recognized the 72-89 epitope and a second encephalitogenic epitope contained in residues 87-99 but not the non-encephalitogenic 43-68 epitope. Unlike those from LN, BP-specific T cell lines and clones derived from the CNS appear to recognize only encephalitogenic epitopes, including epitopes not recognized by LN lines.     

B7-2(CD86)协调刺激因子在急性实验性变态反应性脑脊髓炎(AEAE)发挥主导作用

目的　探讨协调刺激分子B7 1(CD80 )和B7 2 (CD86)在急性EAE发病过程中的作用。方法　观察抗B7 1和B7 2抗体在体外对淋巴细胞抗原特异性增殖反应和细胞因子分泌的抑制作用和在体内对EAE发生过程的影响。结果　抗B7 2抗体抑制PLP136 150抗原引起的特异性细胞增殖和IL 2产生 ,抗B7 2抗体处理过的淋巴母细胞诱导轻症被动EAE ;在主动EAE诱导的早期抗B7 2抗体虽能延缓发病时间但加重病情 ,可能与IL 4分泌不足有关 ;当临床EAE首发症状出现时 ,抗B7 2抗体减轻其临床表现。结论　协调刺激分子B7 2在AEAE发生过程中发挥重要作用     

Nonathymulin treatment of multiple sclerosis: double-blind pilot study

We carried out a randomized, double-blind, placebo-controlled trial of Nonathymulin (NT, synthetic serum thymic factor) in patients with evolutive multiple sclerosis (MS) and moderate disability. Forty matched patients were treated with subcutaneous NT or placebo for 6 months and followed for another 6 months. There was no significant difference in treatment and control groups in the Kurtzke. Disability scores, Ambulation Index and Functional Scale. No significant side effects were recorded. NT is not effective in treating evolutive and moderately disabled MS.