细胞因子在实验性自身免疫性脑脊髓炎耐受中的作用

细胞因子(CK)在实验性自身免疫性脑脊髓炎(EAE)的免疫机制中起着重要作用。Th细胞的不同转化决定EAE的发生、发展或抑制。由多种CK构成的免疫调节网络操纵着Th细胞的免疫应答。通过作用Th细胞使之向抑制EAE方向转化,从而寻找对EAE耐受的途径,是目前EAE研究的一个重要方面。以下就与EAE耐受相关的CK研究进行综述,探讨EAE免疫病理机制。     

胸腺五肽治疗实验性自身免疫性脑脊髓炎的实验研究

目的 应用胸腺五肽(thymopentin,TP-5)干预实验性自身免疫性脑脊髓炎(experiment autoimmue encephalomyelitis,EAE)大鼠,探讨该药物对EAE的治疗作用及其机制.方法 以豚鼠全脊髓匀浆(guinea pig spinal cord homogenate,GPSCH)与完全弗氏佐剂(CFA)为抗原免疫Wistar大鼠建立EAE模型.Wistar大鼠随机分为正常对照组、EAE组、地塞米松(dexamethasone,DXM)组、TP-5小剂量组、TP-5大剂量组.采用双抗体夹心ELISA法检测Wistar大鼠免疫后7、14、21 d不同时间点血清中IL-12、IL-10的含量.结果 与EAE组和TP-5大剂量组比较,TP-5小剂量组和DXM组大鼠的发病率和临床评分显著性降低(P＜0.01);DXM组大鼠的发病率和临床评分低于TP-5小剂量组(P＜0.01).各个时间点EAE组、DXM组、TP-5小剂量组、TP-5大剂量组IL-12含量均较正常对照组明显升高(P＜0.01),免疫后14、21 d DXM组和TP-5小剂量组IL-12水平比EAE组低(P＜0.01);免疫后14、21 dDXM组、TP-5小剂量组IL-10水平与其余3组比较明显升高(P＜0.01).结论 TP-5对EAE有保护作用,其作用机制可能与上调IL-10水平以及下调IL-12水平有关,通过双向调节作用逆转TH1/TH2失衡.     

Wistar大鼠实验性自身免疫性脑脊髓炎模型性别差异研究

目的:分别建立雌性和雄性Wistar大鼠的实验性自身免疫性脑脊髓炎(EAE)模型,并研究两者的差别。方法:应用非灌注法制备的豚鼠全脊髓匀浆分别免疫雌性和雄性两组Wistar大鼠,诱导建立EAE模型,观察30天,取脊髓组织石蜡包埋,病理切片,HE染色,光镜观察。观察两组大鼠在发病时间、发病率、神经功能评分及病程等方面的区别。结果:雌性Wistar大鼠的平均发病时间(13.67±3.50)天,发病率为60%,病程有复发-缓解的特点,神经功能高峰期评分(2.20±1.96)分;雄性Wistar大鼠的平均发病时间(12.18±1.55)天,发病率为85%,呈一过性发病,神经功能高峰期评分(3.46±1.61)分。结论:雄性Wistar大鼠的EAE模型较雌性Wistar大鼠具有发病率高、单相急性病程等特点,为研究多发性硬化发病机制及其发病的性别差异奠定了一定的基础。     

银杏提取物对实验性自身免疫性脑脊髓炎小鼠的影响

目的:探讨银杏提取物(GBE)对实验性自身免疫性脑脊髓炎(EAE)小鼠炎症脱髓鞘病变的影响。方法:应用髓鞘少突胶质细胞糖蛋白33-55(MOG33-55)配以完全弗氏佐剂(CFA)免疫小鼠,诱发EAE模型。将小鼠分为CFA对照组、EAE模型组和GBE治疗组(每日腹腔注射GBE70mg/kg)。通过神经功能评分、行为学实验以及免疫荧光染色,观察GBE对EAE小鼠的影响。结果:GBE组小鼠各时间段神经功能评分均低于EAE组(P0.05),行为学检测显示发病高峰期falling latency时间较EAE组延长10s;GBE组较EAE组视神经髓鞘碱性蛋白(MBP)表达水平增高,可见MBP阳性髓鞘结构包绕轴突;海马伞矢状切片免疫荧光染色证实GBE组CD11b阳性小胶质细胞较EAE组明显减少,但是GFAP阳性星形胶质细胞数量与EAE组无明显差别。结论:GBE可能通过抑制小胶质细胞激活从而延缓EAE小鼠脱髓鞘进程,提示GBE对多发性硬化具有一定的治疗作用。     

在实验性自身免疫性脑脊髓炎模型中探讨MS？…

多发性硬化是一种较常见的中枢神经系统脱髓鞘疾病，Ｔ细胞介导免疫在其发病机制中起一定作用。该病的免疫耐受治疗是目前的研究热点之一，本文综述了近几年在其动物模型实验性自身免疫性脑脊髓炎中进行的研究工作的某些方法和进展。     

降纤药治疗实验性自身免疫性脑脊髓炎小鼠的机制

目的:研究降纤药巴曲酶对实验性自身免疫性脑脊髓炎(EAE)小鼠的预防及治疗作用,初步探讨纤维蛋白沉积在多发性硬化(MS)中的作用机制。方法:采用MOG35-55免疫的C57雌性小鼠,诱发EAE动物模型,分别在免疫后立即(预防组)及免疫后出现症状(治疗组)隔日连续给予降纤药巴曲酶注射至实验结束,观察发病情况并进行临床症状评分。于免疫后30天、40天及60天分别将预防组、治疗组及EAE对照组小鼠脊髓与小脑组织取材后进行组织病理学与免疫组化染色分析,Western blot及实时荧光定量PCR技术观察药物干预对炎性浸润、脱髓鞘、纤维蛋白沉积及胶质细胞活化的影响。结果:降纤预防组及治疗组均可明显减轻EAE小鼠的发病症状、降低临床评分。预防组和治疗组较EAE未用药对照组小鼠炎性细胞浸润明显减少、髓鞘脱失及胶质细胞活化减轻,但轴索损害的改善作用不明显。免疫组化及免疫荧光显示,预防组和治疗组较EAE未用药对照组的髓鞘碱性蛋白(MBP)表达升高而胶质纤维酸性蛋白(GFAP)表达降低,Western blot结果示MBP蛋白表达升高而p-Akt表达降低。实时荧光定量PCR技术也证实了预防及治疗组MBP的mRNA表达升高,组织型纤溶酶原激活物(-tPA)的mRNA表达也升高。结论:在EAE的发病过程中纤维蛋白沉积发挥了重要作用,巴曲酶通过有效降低EAE小鼠体内的纤维蛋白从多个方面改善临床症状和发病过程。     

CM-DiI标记的BMSCs在实验性自身免疫性脑脊髓炎大鼠的体内示踪分布

目的观察并探讨CM-DiI标记骨髓间充质干细胞(BMSCs)移植到实验性自身免疫性脑脊髓炎(EAE)大鼠体内的示踪分布。方法全骨髓培养法分离培养大鼠BMSCs;用CM-DiI荧光染料进行体外标记;将标记后的BMSCs移植到EAE大鼠体内,观察移植后细胞的形态及分布。结果荧光显微镜下可在移植BMSCs的EAE大鼠大脑、小脑和脊髓切片内检测到发出红色荧光的细胞,疾病高峰期数量较多,主要位于软膜下和血管周围,形状为圆形或椭圆形;疾病缓解期大部分细胞仍存在,荧光强度略有减弱。结论 CM-DiI是一种短中期标记、示踪BMSCs的良好染料,CM-DiI标记的BMSCs在EAE大鼠体内主要分布在血管周围,少量向脑实质内迁移。     

在实验性自身免疫性脑脊髓炎模型中探讨MS的免疫耐受治疗

多发性硬化是一种较常见的中枢神经系统脱髓鞘疾病 ,T细胞介导免疫在其发病机制中起一定作用。该病的免疫耐受治疗是目前的研究热点之一 ,本文综述了近几年在其动物模型实验性自身免疫性脑脊髓炎中进行的研究工作的某些方法和进展。     

Regulation of experimental autoimmune encephalomyelitis by chemokines and chemokine receptors

Experimental autoimmune encephalomyelitis (EAE) isa, T cell mediated demyelinating disease of the central nervous system (CNS) that serves as a model for multiple sclerosis (MS). In sights into the pathogenesis of this model may help scientists understand the human disease and aid in rational drug discovery. In this review we summarize the role of chemokines and chemokine receptors in disease pathogenesis and suggest a pathway of events that leads to demyelination and subsequent clinical disease manifestation.     

The effects of intradermal vaccination with DNA encoding for the T-cell receptor on the induction of experimental autoimmune encephalomyelitis in B10.PL mice

Intradermal gene administration was found to induce a more profound immune response than direct intramuscular gene injection. We performed intradermal vaccination of B10.PL mice with DNA encoding for the V 8.2 region of the T-cell receptors (TCR). Three weeks later, these mice were immunized with rat myelin basic protein (MBP). Daily mean clinical scores and mortality rate were lower in this group compared with controls. The proliferative responses of lymph node cells to rat MBP were slightly less in the vaccination groups than in the control groups (p < 0.05). However, we detected no differences between the two groups with regard to the production of MBP-specific IgG, IgG1, & IgG2a antibodies. The levels of cytokine mRNA expression in the vaccination groups were observed higher than in the control groups without antigen-specific stimulation, but all of cytokine expressions between the vaccination and control groups after antigen-specific stimulation were identical. These results demonstrate that intradermal DNA vaccines encoding for TCR might prove to be useful in the control of autoimmune disease.     

霉酚酸酯对实验性自身免疫性脑脊髓炎大鼠症状及血CD4+CD45RA+T淋巴细胞的影响

目的： 研究霉酚酸酯(MMF)对实验性自身免疫性脑脊髓炎(EAE)大鼠症状及血CD4+CD45RA+T淋巴细胞的影响。方法: 用豚鼠全脊髓匀浆和完全弗氏佐剂制成的完全抗原免疫Wistar大鼠制备EAE大鼠模型,以生理盐水和完全弗氏佐剂注射Wistar大鼠作为对照。造模成功且存活的大鼠为25只。随机分为4组：MMF大剂量组（30 mg·kg-1·d-1）、MMF小剂量组（20 mg·kg-1·d-1）、 甲基强的松龙组（30 mg·kg-1·d-1）、EAE组（生理盐水）,并给予相应的药物治疗14 d。每天进行神经功能评分,14d后流式细胞术观察各组血CD4+CD45RA+T淋巴细胞百分率,HE染色观察脑、脊髓组织病理变化。结果: 与对照组相比EAE模型大鼠脑脊髓组织血管周围有大量炎细胞浸润,神经功能评分明显增加,血CD4+CD45RA+T淋巴细胞百分数明显减少(均P<0.01);与EAE组相比,经MMF及甲基强的松龙治疗的各组大鼠神经功能评分下降,病理改变程度减轻,血CD4+CD45RA+T淋巴细胞百分数增加(均P<0.01);且大剂量MMF组疗效优于小剂量MMF组和甲基强的松龙组(均P<0.01);小剂量MMF组和甲基强的松龙组疗效差异无显著。结论: MMF通过上调CD4+CD45RA+T淋巴细胞比例发挥免疫抑制作用,不同剂量间疗效差异显著。     

Exacerbation of experimental autoimmune encephalomyelitis in rodents infected with murine gammaherpesvirus-68

Viral infections have long been suspected to play a role in the pathogenesis of multiple sclerosis. In the present study, two different rodent models of experimental autoimmune encephalomyelitis (EAE) were used to demonstrate the ability of murine gammaherpesvirus-68 (gammaHV-68) to exacerbate development of neurological symptoms. SJL mice received UV-inactivated gammaHV-68 or intranasalgammaHV-68, followed by immunization against proteolipid-protein peptide 139-151. Infected mice became moribund within 10 days post-immunization, whereas mice exposed to UV-inactivated gammaHV-68 recovered. In the second model, Lewis rats were exposed to UV-inactivated gammaHV-68 or to gammaHV-68, followed by passive transfer of encephalitogenic T lymphocytes specific for myelin basic protein. Consistently, infected rats had higher clinical scores, and this result was observed during acute or latent gammaHV-68 infection. It is unlikely that this gammaHV-68-induced exacerbation was due to significant viral replication within the central nervous system since nested PCR, viral plaque assays, and infectious-centers assays demonstrated no detectable virus in spinal cords or brains of infected rodents undergoing EAE. Taken together, these studies demonstrate increased clinical symptoms of EAE in rodents infected by a gammaherpesvirus that has a limited ability to invade the central nervous system.     

Transplantation of autoimmune regulator-encoding bone marrow cells delays the onset of experimental autoimmune encephalomyelitis

The autoimmune regulator (AIRE) promotes "promiscuous" expression of tissue-restricted antigens (TRA) in thymic medullary epithelial cells to facilitate thymic deletion of autoreactive T-cells. Here, we show that AIRE-deficient mice showed an earlier development of myelin oligonucleotide glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis (EAE). To determine the outcome of ectopic Aire expression, we used a retroviral transduction system to over-express Aire in vitro, in cell lines and in bone marrow (BM). In the cell lines that included those of thymic medullary and dendritic cell origin, ectopically expressed Aire variably promoted expression of TRA including Mog and Ins2 (proII) autoantigens associated, respectively, with the autoimmune diseases multiple sclerosis and type 1 diabetes. BM chimeras generated from BM transduced with a retrovirus encoding Aire displayed elevated levels of Mog and Ins2 expression in thymus and spleen. Following induction of EAE with MOG(35-55), transplanted mice displayed significant delay in the onset of EAE compared with control mice. To our knowledge, this is the first example showing that in vivo ectopic expression of AIRE can modulate TRA expression and alter autoimmune disease development.     

The protective effect of total glucosides of white paeony capsules on experimental autoimmune encephalomyelitis

AimsTo learn about the effect and mechanism of total glucosides of white peony capsule (TGP), on experimental autoimmune encephalomyelitis (EAE), an acknowledged animal model of multiple sclerosis (MS).MethodsThe rat model of EAE was induced by subcutaneous injection with guinea pig spinal cord homogenate. The severity of the disease model was assessed by clinical score, hematoxylin and eosin (H&E) and luxol fast blue (LFB). Immunohistochemical assay was used to observe the types of inflammatory cells and adhesive molecule expression. Enzyme-linked immunosorbent assay (ELISA) was applied to detect content of the stem cell growth factor / mast cell growth factor (scf/MGF), interleukin-6 (IL-6) and IL-2. Immunofluorescence assay was applied to observe the expression of connexin43 (Cx43), glial fibrillary acidic protein (GFAP), connexin47 (Cx47) and the monoclonal antibody anti-adenomatous polyposis coli (APC) clone CC1.ResultsCompare with the animals in EAE model group, TGP treated rats (particularly those treated with high doses) showed a significant decrease in morbidity, clinical scores, CNS infiltration of inflammatory cells (including mononuclear macrophages, CD4⁺ and CD8⁺ T cells) and demyelination. The key adhesion molecule ICAM-1, cytokines IL-2、IL-6 and scf/MGF were significantly decreased with TGP treatment. Oppositely, PD-1, connexin47 in oligodendrocytes and connexin43 in astrocytes were elevated with TGP treatment.ConclusionTo sum up, TGP exhibited a significantly prevention and treatment effect on EAE rat model, and this improvement was achieved through a combination way composed of glial and inflammatory cells, junction proteins, various factors including adhesion factors, interleukins and scf/MGF.     

实验性自身免疫性脑脊髓炎脑干听觉通路损害的研究

目的： 探讨实验性自身免疫性脑脊髓炎（EAE）大鼠听觉通路损害的情况，为临床研究提供实验依据。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，采用诱发电位仪检测实验组与对照组的脑干听觉诱发电位，通过脑干及耳蜗神经核HE染色及Weil髓鞘染色从解剖病理的角度证实脑干及耳蜗神经核损害。通过大鼠行为学的变化及光镜下脑和脊髓的病理改变来确定EAE的成功。结果： EAE大鼠的BAEP改变：左耳：Ⅳ、Ⅴ波潜伏期及Ⅰ-Ⅴ波峰间伏期IPL长于对照组（P<0.05）,右耳：Ⅰ、Ⅱ波潜伏期及Ⅰ-Ⅲ Ⅲ-Ⅴ Ⅰ-Ⅴ波峰间伏期IPL长于对照组（P<0.05）；而Ⅲ、Ⅳ、Ⅴ等波的潜伏期显著长于对照组（P<0.01）。病理图像显示脑干耳蜗神经核HE染色有大量淋巴细胞浸润，呈袖套样改变，髓鞘染色显示弥漫性脱髓鞘病变。结论： Wistar大鼠实验性自身免疫性脑脊髓炎（EAE）听神经通路有损害。     

嗅鞘细胞移植对大鼠实验性自身免疫性脑脊髓炎的影响

目的:探索嗅鞘细胞移植实验性自身免疫性脑脊髓炎(EAE)大鼠的移植途径、可行的移植时间窗,移植后的迁移特性以及发挥保护作用的可能机制。方法:分别采用豚鼠脊髓匀浆(GPSCH)与MOGIgd融合蛋白免疫Lewis大鼠,制作EAE模型;每组发病大鼠分别归入:MOG组和GPSCH组。MOG组分为:OECs空白对照组(MOG0组)4只、OECs尾静脉移植组(MOG1组)7只、OECs侧脑室移植组(MOG2组)4只;GPSCH组分为:OECs空白对照组(GPSCH0组)4只、OECs尾静脉移植组(GPSCH1组)4只。发病高峰期,按照实验分组,分别采用立体定向侧脑室细胞移植和尾静脉细胞移植,观察移植后大鼠的临床症状;移植后2周观察OECs在大鼠体内的分布情况及组织病理学方面的缓解情况。结果:OECs分别经尾静脉、侧脑室移植后,EAE大鼠症状改善,与空白对照组神经功能评分峰差值比较有显著差异(F=18.470,P0.01;t=-7.147,P0.01),MOG1组和MOG2组评分峰差值间无显著差异(P0.05);Hoechst33342示踪证实了OECs能在大鼠体内存活及强大的迁移能力;OECs经尾静脉移植后,可以透过破坏的血脑屏障进入脑内,分布于软脑膜和病灶周围;经侧脑室移植的嗅鞘细胞,向病灶局部广泛迁移;组织病理学评分(HE染色和Luxol fast blue髓鞘染色),移植组与未移植组间无显著差异(P0.05),2种途径移植组间亦无显著差异(P0.05)。结论:纯化培养的成年大鼠嗅球嗅鞘细胞分别经尾静脉和侧脑室移植EAE大鼠,均可缓解发病大鼠的症状。