地塞米松对MS的IFN-γ-10分泌性T细胞的影响

目的检测多发性硬化(MS)患者外周血单个核细胞在地塞米松(Dex)影响下的IFN-γ和IL-10的分泌细胞水平.方法采用酶联免疫斑点技术(ELISPOT)检测体外培养的外周血单个核细胞(MNC)在CNS髓鞘素抗原MBP刺激下的地塞米松对照试验,检测IFN-γ和IL-10分泌性T细胞水平,并与其他神经疾病(OND)组及健康对照组的检测结果进行对比.结果显示MS患者IFN-γ分泌细胞水平高于对照组,Dex使MS患者IFN-γ分泌细胞减少,对IL-10分泌细胞无明显影响.结论MS患者存在Th1/Th2细胞因子的失衡,Dex能抑制MSTh1类细胞因子IFN-γ,其治疗作用可能与此有关.     

阿托伐他汀对实验性变态反应性脑脊髓炎T辅助细胞功能的影响

目的探讨阿托伐他汀对实验性变态反应性脑脊髓炎(EAE)豚鼠T辅助细胞功能的影响,探讨阿托伐他汀对EAE发病保护作用的免疫调节机制。方法皮下注射粗制碱性髓鞘蛋白(MBP)建立EAE模型。40只豚鼠分成4组:正常对照组、EAE对照组、EAE低剂量组和EAE高剂量组,每组10只,雌雄各半。ELISA法测定EAE发病高峰期外周血单个核细胞培养上清液的细胞因子INF-γ、IL-4水平。结果EAE对照组IL-4浓度低于正常对照组(P<0.01),EAE高、低剂量组IL-4浓度高于EAE对照组(P<0.01),EAE高剂量组IL-4浓度比EAE低剂量组高(P<0.01)。EAE对照组IFN-γ浓度高于正常对照组(P<0.01),EAE高、低剂量组IFN-γ浓度低于EAE对照组(P<0.01),EAE高剂量IFN-γ浓度比EAE低剂量组低(P<0.01)。结论EAE豚鼠存在Th1细胞的过度活化、Th2细胞分泌活性降低;阿托伐他汀能具有抑制Th1细胞活性、提高Th2细胞分泌能力,具有调节Th1/Th2失衡作用。     

地塞米松对MS的IFN-γ及IL-10分泌性T细胞的影响

目的　检测多发性硬化 (MS)患者外周血单个核细胞在地塞米松 (Dex)影响下的 IFN- γ和 IL- 10的分泌细胞水平。方法　采用酶联免疫斑点技术 (EL ISPOT)检测体外培养的外周血单个核细胞 (MNC)在 CNS髓鞘素抗原 MBP刺激下的地塞米松对照试验 ,检测 IFN -γ和 IL - 10分泌性 T细胞水平 ,并与其他神经疾病 (OND)组及健康对照组的检测结果进行对比。结果　显示 MS患者 IFN- γ分泌细胞水平高于对照组 ,Dex使 MS患者 IFN- γ分泌细胞减少 ,对 IL- 10分泌细胞无明显影响。结论　MS患者存在 Th1/ Th2细胞因子的失衡 ,Dex能抑制 MS Th1类细胞因子 IFN- γ,其治疗作用可能与此有关。     

地塞米松对多发性硬化细胞因子的影响

目的:多发性硬化(MS)是一种由Th1细胞介导的自身免疫性疾病,因而减少T细胞产生γ-干扰素(IFN-γ)或增加白细胞介素-10(IL-10)的产生将会达到治疗MS的目的。通过计数分泌细胞因子IFN-γ和IL一10,检测多发性硬化(MS)患者外周血单个核细胞在地塞米松(Dex)影响下的IFN-γ和IL-10的分泌细胞水平。方法:将外周血单个核细胞暴露于中枢神经系统髓鞘索抗原髓鞘碱性蛋白进行体外短时间培养,然后用Dex作对比试验,用酶联免疫斑点试验(ELISPOT)检测IL-10和IFN-γ分泌细胞,同时检测其它神经病组(OND)及健康对照组。结果:显示MS患者IFN-γ分泌细胞水平高于对照组,Dex使MS患者IFN-γ分泌细胞减少,对IL-10分泌细胞无明显影响。结论:MS患者存在Th1/Th2细胞因子的失衡,Dex能抑制MS Th1类细胞因子IFN-γ,恢复其两类细胞因子之间的平衡而对MS患者产生治疗作用。     

骨髓间充质干细胞干预治疗实验性自身免疫性脑脊髓炎大鼠的研究

目的探讨骨髓间充质干细胞(BMSCs)干预治疗实验性自身免疫性脑脊髓炎(EAE)大鼠的效果及其机制,以及最佳的治疗时机。方法 36只健康雌性SD大鼠随机分为正常对照组、EAE模型组和免疫后当天、第5 d、第10 d、第15 d注入BMSCs干预治疗组(d0、d5、d10、d15 BMSCs组)。用MBP68-86作免疫原制作EAE大鼠模型,分别在免疫后相应时间点静脉注入Wistar大鼠的BMSCs或生理盐水。观察大鼠的发病状况、神经功能缺损评分(NDS)和脑、脊髓组织病理改变。用流式细胞仪检测周围血T细胞亚群,ELISA法检测血干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)、白介素(IL)-4和IL-10水平。结果正常对照组大鼠均正常。与EAE模型组比较,d0 BMSCs组和d5 BMSCs组大鼠的发病数减少,发病时间延迟,高峰期和平均NDS降低,外周血CD4+T细胞明显降低,Treg细胞明显升高,IFN-γ和TNF-α水平明显降低,IL-10水平明显升高(P0.05~0.01),脑、脊髓组织的病变明显减轻;d10 BMSCs组CD4+/CD8+比值明显降低(P0.05)。结论预先用BMSCs干预治疗EAE大鼠有显著的效果,其机制可能为BMSCs通过上调IL-10表达来调节T细胞的增殖活化及功能,抑制异常免疫反应。而在发病后给予BMSCs则无明显效果。     

T细胞疫苗接种对实验性自身免疫性脑脊髓炎大鼠免疫机制的调节作用

目的　探讨 T细胞疫苗 ( TCV)接种对实验性自身免疫性脑脊髓炎 ( EAE)免疫机制的调节作用。方法　取正常及 EAE大鼠腹股沟淋巴结细胞 ,经 MBP抗原诱导 ,制备 MBP特异的 TCV用于接种 ,以 HE染色观察髓鞘病变 ,MTT法检测细胞毒反应 ,FACS方法检测 T细胞亚群 ,ELISA方法检测血清中细胞因子含量。结果　接种 TCV后 ,CD8 细胞百分率上升 ,T细胞对脑细胞的杀伤率下降 ,血清中 IFN-γ与 TNF-α含量下降以及脑髓质炎症反应减弱 ,EAE发病率下降。结论　特异性 TCV接种可降低自身免疫反应性。 TCV通过淋巴细胞亚群的变化及细胞因子的调节 ,发挥对 EAE的免疫预防和治疗效应     

甲基强的松龙与硫唑嘌呤联合治疗对重症肌无力临床和免疫功能的影响

目的　通过检测重症肌无力 ( MG)患者乙酰胆碱受体 ( ACh R)特异性 IFN-γ、IL-1 0分泌细胞水平 ,观察大剂量甲基强的松龙 ( MP)与硫唑嘌呤 ( AZ)联合治疗对 MG患者 Th1及 Th2类细胞因子 ( CK)的影响。方法　用酶联免疫斑点 ( ELISPOT)技术检测 2 1例 MG患者免疫治疗前后外周血 ACh R特异性 IFN-γ、IL-1 0分泌细胞的数目。同时进行临床绝对评分。结果　 MP与 AZ联合治疗可使 MG患者外周血 ACh R特异性 IFN-γ分泌细胞的数目减少 ,对 IL-1 0分泌细胞影响不大。IFN-γ分泌细胞水平与临床绝对评分呈明显正相关 ( r=0 .5 1 2 ,P <0 .0 1 )。结论　 MP与 AZ联合治疗可抑制 MG患者 Th1类 CK的产生 ,恢复 Th1和 Th2类细胞因子之间的平衡 ,继而改善临床症状。 IFN-γ分泌细胞水平可作为反映肌无力严重程度的指标。     

多发性硬化患者周围血IFN-γ和IL-10分泌细胞数的改变

目的:观察多发性硬化(MS)患者分泌γ-干扰素(IFN-γ)及白介素-10(IL-10)的细胞数的变化。方法:采用酶联免疫斑点技术比较MS患者、其他神经疾病患者和健康对照组外周血中髓鞘碱性蛋白(MBP)、乙酰胆碱受体(AChR)、植物血凝素(PHA)反应性及自发性分泌IFN-γ和IL-10的细胞数。结果:MS患者周围血单个核细胞中自发性分泌IFN-γ的细胞数和自发性分泌IL-10的细胞数多于其他神经系统疾病患者和健康对照组。经过MBP刺激后,MS患者分泌IFN-γ和IL-10的细胞数明显多于其他神经系统疾病患者和健康对照组。MS患者周围血细胞对MBP存在特异性反应。MS患者分泌IFN-γ的细胞数多于分泌IL-10的细胞数。结论:Th1类细胞因子(IFN-γ)分泌大于Th2类细胞因子(IL-10)与MS活动密切相关。MS患者的淋巴细胞对MBP存在特异性反应。     

Lewis大鼠主动性实验性变态反应性脑脊髓炎T淋巴细胞增殖和肿瘤坏死因子-α变化的意义

目的 :探讨实验性变态反应性脑脊髓炎 ( EAE) T淋巴细胞增殖和肿瘤坏死因子 -α变化的意义。方法 :观察L ewis大鼠主动 EAE临床症状、 T淋巴细胞增殖以及血清肿瘤坏死因子 ( TNF) p-α水平。结果 :EAE临床发病过程中 ,T淋巴细胞增殖活跃 ,血清 TNF- α水平异常升高 ,并与临床症状呈正比。结论 :研究表明 :EAE时 ,T淋巴细胞增殖旺盛和 TNF- α生成增多 ,进而导致血脑屏障 ( BBB)的破坏 ,炎细胞浸润 ,从而诱发 EAE     

重症肌无力患者血清Th1/Th2/Th17细胞因子的变化及意义

目的：分析重症肌无力（MG）患者血清CD4^＋ T细胞主要细胞因子的水平,探讨不同亚型CD4^＋ T细胞分泌的细胞因子在MG发病机制中的作用。方法：用ELISA测定93例MG患者和34名健康对照者血清中各项细胞因子（IL-2、IL-12、IFN-γ、TNF-α、IL-4、IL-10、IL-13和IL-17）的水平,分组行统计学分析。结果：与健康对照组相比,MG患者Th1细胞相关各细胞因子（IL-2、IL-12、IFN-γ及TNF-α）均明显升高,差异有统计学意义（P〈0.05）;Th2细胞相关的细胞因子IL-4、IL-10差异无统计学意义（P〉0.05）,仅IL-13水平升高;Th17细胞的细胞因子IL-17水平差异无统计学意义（P〉0.05）。MG眼肌型与全身型患者血清中各细胞因子水平的差异无统计学意义（P〉0.05）,在不同病程的MG患者中差异也无统计学意义（P〉0.05）。结论：Th1细胞因子在MG发病机制中发挥重要作用,而Th2细胞及其细胞因子在MG机制中的角色各异。     

Infiltration of Th1 and Th17 cells and activation of microglia in the CNS during the course of experimental autoimmune encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) is a mouse model for multiple sclerosis, where disease is mediated by autoantigen-specific T cells. Although there is evidence linking CD4⁺ T cells that secrete IL-17, termed Th17 cells, and IFN-γ-secreting Th1 cells with the pathogenesis of EAE, the precise contribution of these T cell subtypes or their associated cytokines is still unclear. We have investigated the infiltration of CD4⁺ T cells that secrete IFN-γ, IL-17 or both cytokines into CNS during development of EAE and have examined the role of T cells in microglial activation. Our findings demonstrate that Th17 cells and CD4⁺ T cells that produce both IFN-γ and IL-17, which we have called Th1/Th17 cells, infiltrate the brain prior to the development of clinical symptoms of EAE and that this coincides with activation of CD11b⁺ microglia and local production of IL-1β, TNF-α and IL-6 in the CNS. In contrast, significant infiltration of Th1 cells was only detected after the development of clinical disease. Co-culture experiments, using mixed glia and MOG-specific T cells, revealed that T cells that secreted IFN-γ and IL-17 were potent activators of pro-inflammatory cytokines but T cells that secrete IFN-γ, but not IL-17, were less effective. In contrast both Th1 and Th1/Th17 cells enhanced MHC-class II and co-stimulatory molecule expression on microglia. Our findings suggest that T cells which secrete IL-17 or IL-17 and IFN-γ infiltrate the CNS prior to the onset of clinical symptoms of EAE, where they may mediate CNS inflammation, in part, through microglial activation.     

Cytokine production in the central nervous system of Lewis rats with experimental autoimmune encephalomyelitis: dynamics of mRNA expression for interleukin-10, interleukin-12, cytolysin,tumor necrosis factor α and tumor necrosis factor β

The kinetics of mRNA expression in the central nervous system (CNS) for a series of putatively disease-promoting and disease-limiting cytokines during the course of experimental autoimmune encephalomyelitis (EAE) in Lewis rats were studied. Cytokine mRNA-expressing cells were detected in cryosections of spinal cords using in situ hybridization technique with synthetic oligonucleotide probes. Three stages of cytokine mRNA expression could be distinguished: (i) interleukin (IL)-12, tumor necrosis factor (TNF)-β (=lymphotoxin-α) and cytolysin appeared early and before onset of clinical signs of EAE; (ii) TNF-α peaked at height of clinical signs of EAE; (iii) IL-10 appeared increasingly at and after clinical recovery. The early expression of IL-12 prior to the expression of interferon-γ (IFN-γ) mRNA shown previously is consistent with a role of IL-12 in promoting proliferation and activation of T helper 1 (Th1) type cells producing IFN-γ. The TNF-β mRNA expression prior to onset of clinical signs favours a role for this cytokine in disease initiation. A pathogenic effector role of TNF-α was suggested from these observations that TNF-α mRNA expression roughly paralleled the clinical signs of EAE. This may be the case also for cytolysin. IL-10-expressing cells gradually increased to high levels in the recovery phase of EAE, consistent with a function in down-regulating the CNS inflammation. From these data we conclude that there is an ordered appearance of putative disease-promoting and -limiting cytokines in the CNS during acute monophasic EAE.     

Cytokine phenotype of human autoreactive T cell clones specific for the immunodominant myelin basic protein peptide (83–99)

Experimental allergic encephalomyelitis (EAE), an animal model resembling multiple sclerosis (MS), is mediated by myelin antigen-specific CD4+ T cells secreting cytokines such as interferon-γ (IFN-γ), tumor necrosis factor-β (TNF-β), and the proinflammatory cytokine TNF-α—all associated with the T-helper-1 (Th1) T cell subset. Based on numerous similarities between MS and EAE, it has been postulated that Th1-like T cells are involved in the pathogenesis of MS. Production of proinflammatory cytokines such as IFN-γ and, in particular, TNF-α/β by autoreactive T cells is considered crucial for the initiation and amplification of inflammatory brain lesions and possibly also for direct myelin damage. In contrast, regulatory cytokines such as interleukin-4 (IL-4), IL-10, and IL-13, which are associated with the Th2-like phenotype, may play a role in the resolution of relapses. Although the human T cell response to myelin basic protein (MBP) is well characterized in terms of antigen specificity, HLA restriction, and T cell-receptor (TCR) usage, little is known about the cytokine pattern of these autoreactive T cells. To gain such information, conditions for studying cytokine secretion by human autoreactive T cell clones (TCC) were established. The cytokine secretion profile of human autoreactive CD4+ TCC, specific for myelin basic protein peptide (83–89) [MBP(83–99)], a candidate autoantigen in MS, was investigated. Our results show that TCC cytokine production in long-term culture was stable. In addition, the correlation of various cytokines within specific TCC revealed differences compared to murine T cells. The comparison of 30 human MBP(83–99)-specific TCC demonstrated heterogeneity in cytokine secretion, with a continuum between Th1- and Th2-like cells rather than distinct Th1 or Th2 subsets. These data are important for further investigation of the potential role of cytokines in the inflammatory process of MS, and provide a powerful tool to investigate therapeutic interventions with respect to their influence on cytokine secretion of autoreactive T cells. © 1996 Wiley-Liss, Inc.     

Ingested (oral) ACTH inhibits EAE

Ingested type I IFN and SIRS peptide inhibit EAE. We examined whether another immunoactive protein, ACTH, would have similar anti-inflammatory effects in EAE after oral administration. B6 mice were immunized and gavaged with control saline or ACTH starting on the onset of disease. ACTH decreased clinical score and decreased inflammatory foci. CNS lymphocytes showed decreases in IL-17 (T(eff)) and Th1-like encephalitogenic cytokines IL-2 and IFN-γ in the ACTH fed group compared to the mock fed group. Adoptive transfer of ACTH fed splenocytes into MOG immunized recipient mice with early clinical disease suppressed disease severity compared to splenocytes from mock fed donors. The protected recipients showed decreased splenic IL-17 (T(eff)) and Th1-like cytokine IFN-γ and increased CNS secretion of immunoregulatory IL-4 and chemokine M-CSF. Splenic CD4+CD25+ FoxP3+ frequency doubled in ACTH fed compared to control fed mice. Increased immuno-regulatory IL-4 and M-CSF secreting cell populations is the mechanism of protection in adoptively protected recipients and reflects the direct action of ACTH on the immune system.     

Role of IL-10 −1082, IFN-γ +874, and TNF-α −308 Genes Polymorphisms in Suicidal Behavior

In this study, it was determined whether the IL-10 ?1082, IFN-γ +874, and TNF-α ?308 polymorphisms were associated with suicidal behavior. One hundred forty five patients with suicidal behavior and 160 normal individuals were genotyped for IL-10 ?1082, IFN-γ +874, and TNF-α ?308 polymorphisms using ASO-PCR method. TNF-α ?308 G/G genotype has been increased in males with completed suicide behavior versus control group (p value = 0.017). IL-10 ?1082 A/A genotype is higher in both male and female suicide completed groups (p value = 0.017). IFN-γ (+874) A/A genotype was significantly higher in males with completed suicide behavior versus normal male control (p value = 0.027). It can be concluded that IL-10, IFN-γ, and TNF-α polymorphisms may play a role in suicidal behavior.     

Pentoxifylline, a phosphodiesterase inhibitor, induces immune deviation in patients with multiple sclerosis

The outcome of immune responses can be predicted by the lymphokine production pattern of the participating cells. Cytokines of the T helper type 1 (Th1) cells mediate inflammatory responses and delayed-type hypersensitivity (DTH), whereas Th2-like T cells predominantly produce cytokines, which stimulate antibody production by B cells. Immunoregulatory therapy of autoimmune diseases with unknown antigens may be achieved by inhibiting the production of inflammatory cytokines and induction of protective cytokines of Th2-like T cells. To determine the immunoregulatory capacity of the phosphodiesterase inhibitor pentoxifylline (PTX), which is known to suppress the production of tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ), this drug was used in mitogen and antigen-stimulated lymphocyte cultures as well as in patients with multiple sclerosis. PTX significantly decreased TNF-α and interleukin-12 (IL-12), whereas it increased IL-4 and IL-10 production. In addition, PTX inhibited cell proliferation, which was associated with a marked reduction in CD25 (IL-2 receptor α-chain) and CDS4 (intercellular adhesion molecule-1; ICAM-1) expression. Increasing doses of PTX significantly reduced TNF-α and IL-12 mRNA expression of blood mononuclear cells, but increased IL-4 and IL-10 expression in eight patients with relapsing-remitting multiple sclerosis. These results indicate that PTX modulates immune reactions favouring a Th2-like response and may therefore be useful for the treatment of autoimmune diseases with a dominant Th1-like T cell response.     

Cytokines and the pathogenesis of myasthenia gravis

Myasthenia gravis (MG) and its animal model experimental autoimmune myasthenia gravis (EAMG) are caused by autoantibodies against nicotinic acetylcholine receptor (AChR) in skeletal muscle. The production of anti-AChR antibodies is mediated by cytokines produced by CD4⁺ and CD8⁺ T helper (Th) cells. Emerging investigations of the roles of cytokines in MG and EAMG have revealed that the Th2 cell related cytokine interleukin 4 (IL-4), an efficient growth promoter for B-cell proliferation and differentiation, is important for anti-AChR antibody production. IL-6 and IL-10 have similar effects. The Th1 cytokine IFN-γ is important in inducing B-cell maturation and in helping anti-AChR antibody production and, thereby, for induction of clinical signs and symptoms. Results from studies of time kinetics of cytokines imply that IFN-γ is more agile at the onset of EAMG, probably being one of the initiating factors in the induction of the disease, and IL-4 may be mainly responsible for disease progression and persistance. Even though other Th1 cytokines like IL-2, tumor necrosis factor α (TNF-α), and TNF-β as well as the cytolytic compound perforin do not directly play a role in T-cell-mediated help for anti-AChR antibody production, they are actually involved in the development of both EAMG and MG, probably by acting in concert with other cytokines within the cytokine network. In contrast, transforming growth factor β (TGF-β) exerts immunosuppressive effects which include the down-regulation of both Th1 and Th2 cytokines in MG as well as EAMG. Suppressive effects are also exerted by interferon α (IFN-α). Based on elucidation of the role of cytokines in EAMG and MG, treatments that up-modulate TGF-β or IFN-α and/or suppress cytokines that help B-cell proliferation could be useful to improve the clinical outcome. © 1997 John Wiley & Sons, Inc. Muscle Nerve, 20, 543–551, 1997     

Serum and cerebrospinal fluid cytokine concentrations in subacute sclerosing panencephalitis

Subacute sclerosing panencephalitis (SSPE) is a neurodegenerative disease due to persistent measles virus infection. Its immunopathogenesis is unknown. Tumor necrosis factor (TNF)-α, interleukin (IL)-2, IL-6, IL-10 and IL-4 concentrations were measured in cerebrospinal fluid (CSF) and serum samples from 30 SSPE patients and 19 control subjects by cytometric bead array. CSF and serum IFN-γ, IL-12 and IL-18 levels were measured in 18 SSPE patients by ELISA. Serum IL-4 and IL-10 (p < 0.001), CSF IL-4 (p < 0.001) and IL-6 (p = 0.049) concentrations were lower, and serum IL-2 concentrations, higher (p = 0.001) in SSPE patients. Serum TNF-α and IL-6, CSF TNF-α, IL-10, and IL-2 concentrations were not different between SSPE and control groups. Serum IFN-γ levels were higher in stage I and II than stage III patients (p < 0.05), whereas there was no difference between stages in terms of other cytokines. The levels of Th2-type cytokines: IL-4, IL-6 and IL-10 were suppressed in our SSPE cases. This finding, along with relatively elevated IFN-γ and IL-2 levels, may suggest more active effector T cells compared to regulatory T cells (Treg), especially induced Treg, in early disease. High serum IL-2 concentrations might indicate peripheral Th1 activation. Discrepancies between various reports in the literature should be examined in view of the ages, stage and treatments of the patients studied. The interplay of various cytokines or cellular systems which may vary over time and between patients. Studies of treatment measures favoring the preservation of the early inflammatory response may be of interest in SSPE.