趋化因子mRNA在实验性变态反应性神经炎中表达的研究

目的:实验性变态反应性神经炎(EAN)是一类T细胞介导的周围神经系统的自身免疫病,可用牛坐骨神经加完全氟氏佐剂诱导而成。本文研究趋化因子mRNA在实验性变态反应性神经炎(EAN)中的表达并探索其可能的作用。方法:用兔坐骨神经匀浆免疫Wistar大鼠,诱导格林巴利综合症(GBS)的动物模型EAN;采用地高辛标记的寡核苷酸探针检测EAN病变神经组织浸润细胞上趋化因子单核细胞趋化蛋白-1(MCP-1)及巨噬细胞炎性蛋白-1β(MIP-1β)mRNA表达情况。结果:MCP-1mRNA在临床症状出现前1-2天(14天)水平最高,随后逐渐下降;MIP-1 βmRA在临床症状出现前1-2天水平开始升高,在临床症状达到高峰时(21天)最高,进入恢复期后降至基础水平。结论:趋化因子在EAN的炎性细胞迁移及浸润进入神经细胞过程中起到重要作用。     

Chronic relapsing experimental allergic encephalomyelitis in the Lewis rat

Chronic relapsing experimental allergic encephalomyelitis (CR-EAE) was induced in rats with an emulsion of guinea-pig spinal cord tissue (GPSC) in complete Freund's adjuvant (CFA) enriched with Mycobacterium tuberculosis H37RA (Tbc). 78% of the sensitized rats developed a CR-EAE showing 2 to 3 clinical relapses during the first 40 days. After 60-80 days, approximately half of the rats with CR-EAE had a further relapse which was followed by complete recovery in only 35% of the cases. The remaining 65% of these animals showed a progressive state of the disease, characterized by paralysis or severe motor deficit, eventually leading to death. CR-EAE in rats showed some similarities to multiple sclerosis in man (MS) and it may be a useful model for the study of this disease.     

Erythropoietin exerts an anti-inflammatory effect on the CNS in a model of experimental autoimmune encephalomyelitis

In recent work we reported that systemically administered erythropoietin (EPO) crosses the blood-brain barrier and has protective effects in animal models of cerebral ischemia, brain trauma and in a rat model of experimental autoimmune encephalomyelitis (EAE). Here we characterize the effect of systemic EPO on the inflammatory component of actively induced, acute EAE in Lewis rats. Administration of EPO at doses of 500-5000 U/kg bw i.p., daily from day 3 after immunization with myelin basic protein (MBP), delayed the onset of EAE and decreased its clinical score at peak time (days 12-13). Immunohistochemical analysis of the spinal cord using anti-glial fibrillary acidic protein (GFAP) and anti-CD11b antibodies showed that EPO markedly diminished inflammation and glial activation/proliferation. EAE induced significant levels of TNF and IL-6 in the spinal cord, where IL-6 was maximum at the onset of the disease (day 10) and TNF at its peak (day 12). EPO delayed the increase of TNF levels, without altering their peak levels, and markedly reduced those of IL-6 suggesting that the decreased inflammation and clinical score may be in part upon attenuation of IL-6. On the other hand, EPO was without effect in a model of adjuvant-induced arthritis in Lewis rats, suggesting a specificity towards autoimmune demyelinating diseases. These data suggest that EPO might act as a protective cytokine in inflammatory pathologies of the CNS.     

Chronic relapsing experimental allergic encephalomyelitis: CNS plaque development in unsuppressed and suppressed animals

Summary Central nervous system (CNS) lesion morphology has been studied in inbred Strain 13 guinea pigs sensitized for chronic relapsing EAE in which the disease was either left to develop (unsuppressed) or was suppressed with injections containing myelin basic protein (MBP). Pathologic changes correlated well with clinical activity. In unsuppressed chronic EAE animals, active clinical disease was invariably matched by acute inflammation in the CNS. In more chronic states, the CNS displayed fibrosis and remyelination while response showed the CNS to contain recent changes superimposed upon old lesions. In animals in which the disease was suppressed by injections of MBP, clinical signs did not develop. However, some early subclinical changes were seen morphologically. These lesions were able to remyelinate early on and there was no progression in lesion formation. Apparently, therefore, MBP had a beneficial effect upon the course of the disease and had promoted structural repair. It thus appears that MBP therapy might be one effective approach for the prevention of chronic relapsing EAE. The findings should prove relevant to future MBP trials in multiple sclerosis.     

Restraint stress suppresses experimental allergic encephalomyelitis in Lewis rats

We studied the effect of restraint stress on experimental allergic encephalomyelitis (EAE), a model for the human disease multiple sclerosis (MS), in Lewis rats. Rats were subjected to 12 h restraint stress for 3 consecutive nights from the first (day 1) or the eighth day (day 8) after sensitization with the antigen (guinea-pig spinal cord). All controlled rats exhibited clinical and histologie signs of EAE. The mean ± SD of incubation time, clinical score (0–4) and histologic score (0–3) for this group were 12.8 ± 1.0 days, 2.7 ± 0.6, and 2.3 ± 0.7, respectively. Restraint stress from day 8 significantly suppressed EAE: the mean ± SD of incubation time, clinical score, and histologic score for this group were 17.2 ± 2.2 days (p < 0.001), 1.8 ± 1.3 (p < 0.05), and 1.5 ± 0.9 (p < 0.02), respectively. Restraint stress from day 1 did not modify EAE. The findings suggest that stressful factors may exert an influence on the clinical course of MS.     

Chemokine expression by astrocytes plays a role in microglia/macrophage activation and subsequent neurodegeneration in secondary progressive multiple sclerosis

The pathological hallmarks of secondary progressive (SP) multiple sclerosis (MS) include slowly expanding demyelination and axonal damage with less inflammation. To elucidate the pathomechanisms of secondary progressive (SP) multiple sclerosis (MS), we have investigated the expression of chemokines, chemokine receptors, matrix metalloproteinase-9 (MMP-9) and immunoglobulins in the demyelinating plaques. Immunohistochemical analysis revealed that numerous hypertrophic astrocytes were observed at the rim, but not in the center, of the chronic active lesions. Microglia/macrophages phagocytosing myelin debris were also found at the lesion border. In contrast, T cell infiltration was minimal in these plaques. Characteristically, at the rim of the lesions, there were abundant immunoreactivities for monocyte chemoattractant protein-1 (MCP-1)/CCL2 and interferon-γ inducible protein-10 (IP-10)/CXCL10 and their receptors, CCR2 and CXCR3, while these immunoreactivities were weak in the center, thus forming a chemokine gradient. Double immunofluorescense staining demonstrated that cellular sources of MCP-1/CCL2 and IP-10/CXCL10 were hypertrophic astrocytes and that both astrocytes and microglia/macrophages expressed CCR2 and CXCR3. MMP-9 was also present at the rim of the lesions. These results suggest that MCP-1/CCL2 and IP-10/CXCL10 produced by astrocytes may activate astrocytes in an autocrine or paracrine manner and direct reactive gliosis followed by migration and activation of microglia/macrophages as effector cells in demyelinating lesions. Targeting chemokines in SPMS may therefore be a powerful therapeutic approach to inhibit lesional expansion.     

实验性变应性猴脑脊髓炎的超微结构改变

目的探讨自身免疫性中枢神经系统（ＣＮＳ）脱髓鞘的病理特点和发病过程。方法建立猴实验性变态反应性脑脊髓炎（ＥＡＥ）的模型，并从病情不同阶段进行病理取材和电镜观察。结果（１）ＥＡＥ脱髓鞘最早的靶是少突胶质细胞（ＯＤＣ），而不是髓鞘本身；（２）无论急性期还是慢性期，脱髓鞘病灶内只有极少的炎细胞浸入；（３）急性期髓鞘改变轻微，以变性为主，ＯＤＣ肿胀显著，轴突相对完整。远离髓鞘变性区有大量炎细胞浸入；（４）慢性期病变区髓鞘脱失明显，ＯＤＣ严重变性或部分丢失，继发性轴突变性，边缘可见少量薄的髓鞘再生，血脑屏障破坏。结论ＥＡＥ的ＣＮＳ脱髓鞘过程首先累及ＯＤＣ。     

Chronic relapsing experimental allergic encephalomyelitis: its value as an experimental model for multiple sclerosis

Summary Comparison of the pathohistology of chronic relapsing experimental allergic encephalomyelitis (CR-EAE) and multiple sclerosis (MS) reveals a close similarity. Thus, CR-EAE appears to be a valuable model for the study of pathogenetic factors leading to the formation of MS lesions, although the induction of the disease may be different (active sensitization with CNS antigens and adjuvant in CR-EAE versus unknown etiology in MS). CR-EAE furthermore mimicks the pathohistological patterns of other related human inflammatory demyelinating diseases (i.e., acute perivenous leukoencephalomyelitis and acute hemorrhagic leukoencephalomyelitis). The expression of an acute, predominantly inflammatory versus chronic inflammatory demyelinating disease in this model depends upon the time interval between sensitization and sampling of the animals. Recent evidence is discussed that a cooperation between cellular and humoral immune mechanisms, directed against multiple CNS antigens, is responsible for the formation of large demyelinated plaques in EAE and MS.Supported by Austrian Science Research Fund, Project S25/07     

慢性实验性变应性猴脑脊髓炎的轴突病变及其意义

目的 探讨自身免疫性中枢神经系统脱髓鞘疾病慢性型的病理点及其临床意义。方法 建立猴实验性变态反应性脑脊髓炎（EAE）模型,于首次发病后4年进行病理取材和电镜观察。结果　①活动性病灶内轴突病变十分突出,其形式包括有空泡样变性、皱缩或消失,此外也可见到成片的髓鞘松解、断裂或融合,以及少突胶质细胞变性,见散在巨噬细胞;②可疑活动性病灶内轴突病变程度稍轻,以空泡样变为主,轴突完全消失及皱缩则少见,部分髓鞘内板松解,亦有少突胶质细胞变性,散在巨噬细胞。结论 慢性EAE的病理改变同时存在髓鞘与轴突的变性,多发性硬化后期不可逆的功能障碍可能与后者有着更大的相关。     

慢性实验性变应性猴脑脊髓炎的超微结构改变

目的：探讨自身免疫性中枢神经系统脱髓鞘疾病慢性型的病理特点及其可能机制。方法：成功建立猴实验性态反应性脑脊髓炎模型数年后，根据MR摄片结果位病灶并作分类，然后进行病理取材和电镜观察超微结构。结果：（1）活动性病灶内成片的髓鞘松解、断裂或融合，轴突空泡样变性，皱缩或消失，少突胶质细胞变性，未见淋巴细胞浸润，仅见散在巨噬细胞，伴明显的间质水肿；（2）可疑活动性病灶内见部分髓鞘内板松解，其内轴突有较度空泡样变，亦见少突胶质细胞变性，散在巨噬细胞，未见淋巴瘤浸润，结论：慢性EAE的病理改变不仅有髓鞘的变性，同时轴突的病变也十分明显。     

Dendritic cells in experimental allergic encephalomyelitis and multiple sclerosis

The mechanisms by which autoimmune diseases are triggered and by which the activation of autoreactive T cells is initiated and maintained are not yet fully understood. As the most potent antigen presenting cells (APC), and also being responsible for antigen transport as well as primary sensitisation of T cells, dendritic cells (DC) are capable of breaking the state of self-ignorance and inducing aggressive autoreactive T cells. In the development of autoimmune diseases, different types of DC exhibit distinct properties for inducing Th1/Th2 cell responses. Appropriate cytokines can convert immunogenic DC to tolerogenic DC. Utilizing the possibility to promote the tolerogenic effects of DC, a new therapeutic tool might soon become available to treat multiple sclerosis and other autoimmune diseases.     

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。     

猴急慢性实验性变应性脑脊髓炎的病理研究

目的　探讨急、慢性猴 (猕猴类 )实验性变应性脑脊髓炎 (EAE)的病理特点。方法　从EAE猴病程不同阶段进行病理取材 ,制成切片 ,用HE、Bielschowsky改良法、Page法、Holzer法染色及间接免疫荧光法观察CD4、CD8和B淋巴细胞。结果　 ( 1)猴急性EAE脑组织炎症反应重 ,分布在小静脉周围形成血管套 ,以CD4淋巴细胞浸入为主。脱髓鞘病灶微小 ,分布弥散 ,轴突改变轻 ,无出血 ,类似急性播散性脑脊髓炎 (ADEM)。 ( 2 )猴慢性EAE脑组织炎症反应轻 ,分布在血管外周 ,以CD8和B细胞为主。脱髓鞘病灶较大 ,界线清晰 ,呈局灶性分布。病变部位轴突变性严重及轴突丧失 ,有胶质斑块形成 ,与慢性多发性硬化 (MS)的活动期病理改变有惊人的相似。结论　支持猴慢性EAE是MS的优秀模型。另外 ,用相同抗原诱导出不同病程的EAE提示ADEM和MS可能是同一疾病的不同病程 ,或可能是同一种病因 ,而致病程不同。     

Serial adoptive transfer of murine experimental allergic encephalomyelitis: successful transfer is dependent on active disease in the donor

In an attempt to understand the mechanisms underlying disease progression in adoptively transferred experimental allergic encephalomyelitis (EAE), and perhaps multiple sclerosis (MS), this study has examined the transfer of EAE serially from primary to secondary and tertiary recipients using myelin basic protein (MBP)-responsive lymphocytes. It was found that EAE could be serially transferred only when there was acute or relapsing disease activity in the donor animal. Cells from donors with quiescent disease did not transfer EAE. Autoradiographic attempts to locate primary adoptively transferred cells in the central nervous system of secondary and tertiary recipients were uniformly unsuccessful. These findings implicate the requirement of effector cell activation in the donor and the recruitment of augmenting autoimmune cells in lesion formation.     

Electrophysiological follow-up of acute and chronic experimental allergic encephalomyelitis in the Lewis rat

Summary Cortical somatosensory evoked potentials (c-SEP) and flash visual evoked potentials (f-VEP) were serially recorded in acute monophasic and chronic relapsing experimental allergic encephalomyelitis (EAE) in the Lewis rat. In acute EAE, a significantly delayed latency and broadened peak of the c-SEP were observed corresponding to the clinical onset, and then returned to normal with the disappearance of clinical signs. In chronic EAE, the c-SEP showed the same changes as in acute EAE, also reflecting the first attack, remission and relapsing phase. However, chronic EAE, when paralysis had recovered in the relapsing phase, showed c-SEP abnormalities suggestive of subclinical active lesions. In contrast, the f-VEP showed no obvious abnormalities in acute or chronic EAE. These findings suggest that the c-SEP is an objective and sensitive index for detecting clinical and pathological changes in acute and chronic EAE in the Lewis rat.     

Long-term treatment of chronic relapsing experimental allergic encephalomyelitis by transforming growth factor-β2

It had been demonstrated previously that the administration of transforming growth factor-β1 (TGF-β1) reduced the clinical severity of experimental allergic encephalomyelitis (EAE). Treatment with the related immunosuppressive molecule, TGF-β2, resulted in similar inhibition of T cell activation and proliferation in vitro. Long-term treatment was affective in reducing clinical severity of EAE and the number of relapses in mice receiving either myelin basic protein- or peptide-91-103-specific T cell lines. When examined histologically, mice that had received TGF-β2 demonstrated significantly less inflammation and demyelination in the central nervous system. Examination of other organs demonstrated to pathology or deleterious side effects from long-term TGF-β2 therapy. These findings have relevance for the use of TGF-β2 as a therapeutic agent for the human demyelinating disease, multiple sclerosis.     

Chemokine alterations in bipolar disorder: A systematic review and meta-analysis

We aimed to perform a systematic review and meta-analysis of studies examining the levels of chemokines in peripheral blood of patients with bipolar disorder (BD) and healthy controls. Meta-analysis was based on random-effects models with Hedges’ g as the effect size estimate. We included 13 eligible studies (1221 BD patients and 663 controls). The following chemokines were analysed: interleukin-8 (IL-8), monocyte-chemoattractant protein-1 (MCP-1), eotaxin-1, eotaxin-2 and interferon-γ-induced protein 10 (IP-10). The levels of IL-8 (N = 8, g = 0.26, 95%CI: 0.11–0.41, p < 0.001), MCP-1 (N = 8, g = 0.40, 95%CI: 0.18–0.63), eotaxin-1 (N = 3, g = 0.55, 95%CI: 0.21–0.89, p = 0.001) and IP-10 (N = 4, g = 0.95, 95%CI: 0.67–1.22, p < 0.001) were significantly higher in BD patients as compared with controls. Subgroup analyses revealed that elevated levels of IL-8 (N = 5, g = 0.75, 95%CI: 0.42–1.07, p < 0.001) and MCP-1 (N = 4, g = 0.57, 95%CI: 0.28–0.86, p < 0.001) appeared only in BD patients during their depressive phase. Illness duration was associated with significantly lower levels of IL-8 in meta-regression analysis. In turn, elevated levels of IP-10 were present during euthymia (N = 2, g = 0.76, 95%CI: 0.43–1.10, p < 0.001) but not depression (N = 2, g = 1.81, 95%CI: −0.16 to 3.77, p = 0.072). The analysis of eotaxin-1 levels was mainly based on studies of euthymic BD patients (N = 3). Our results suggest that chemokine alterations in BD might be related to mood state. Elevated levels of IL-8 and MCP-1 might be specific to depression. Available evidence indicates that increased levels of eotaxin-1 and IP-10 appear in euthymia; however, more studies are needed to address these alterations in other mood states.     

调节性T细胞在实验性变态反应性脑脊髓炎和多发性硬化发病中的作用

调节性T细胞(regulatory T cells,Treg)是一类具有免疫调节功能的T细胞哑群.近年来,Treg细胞在实验性变态反应性脑脊髓炎(experimental allergical encephalomyelitis,EAE)、多发性硬化(multiple sclerosis,MS)发病中的作用越来越受到关注,小鼠Treg细胞缺失可导致特异性自身免疫性疾病,增加Treg细胞的功能可以减轻或抑制EAE.最近的研究结果表明,MS本身也伴随着成熟Treg细胞的受损或功能障碍.