实验性变态反应性神经炎的大鼠淋巴细胞钾离子通道与细胞及体液免疫的关系

目的:观察淋巴细胞K 通道在实验性变态反应性神经炎(EAN)大鼠中的表达情况及其与细胞和体液免疫的关联。方法:RT-PCR测定淋巴结和脾脏细胞Kv1.3和IKCa1的mRNA表达量,流式细胞分析相应细胞的增殖能力,间接ELISA测定特异性抗体水平,计算相关系数。结果:淋巴细胞Kv1.3和IKCa1的表达水平与细胞增殖能力紧密相关,与EAN临床严重程度无关,在一定程度上反应了EAN的总体细胞免疫水平。结论:淋巴细胞K 通道可以在以细胞免疫为主导的自身免疫性疾病中作为一个潜在的干预靶点。     

实验性变态反应性神经炎动物模型的建立和病理特点

参照Ｙ，Ｌｏｎｄｏｎ方法，用牛的鹇坐骨社地髓鞘碱性蛋白（ＭＢＰ），并对所提取的ＭＢＰ进行生化及生物活性鉴定，测得ＭＢＰ的分子量为１４ ４００￣２０ ０００道尔顿之间，用所提取的ＭＢＰ免疫新西兰白兔。观察临床表现并行病理学检查，１０只兔子有７只出现不同程度的临床症状，发病率达７０％，病理检查发现，发病兔炎细胞浸润以颈神经根、腰神经根、马尾神经损害重于坐骨神经于（Ｐ〈０．０１），单神经纤维分离，半薄切     

硫脂诱导的实验性变态反应性多发性神经炎

目的　探讨硫脂在周围神经脱髓鞘损害中的免疫原性。方法　以硫脂免疫豚鼠 ,观察临床、血清学变化 ,并行病理学检查。结果　免疫 14周的 13只豚鼠中有 11只 ( 85 % )血清中出现高滴度抗硫脂IgG抗体 ,且出现程度不同的临床症状。被致敏动物坐骨神经、脊神经根发生脱髓鞘改变 ,有的脊神经根中可见炎性细胞浸润 ;神经鞘膜IgG荧光沉积明显。动物血清中抗硫脂IgG滴度与动物临床分级、坐骨神经脱髓鞘程度呈正相关 (r分别为 0 .914 2和 0 .95 2 7,P均 <0 .0 1)。结论　硫脂是实验性变态反应性多发性神经炎的重要免疫原之一。     

实验性变态反应性神经炎动物模型的建立和病理特点

参照Ｙ．Ｌｏｎｄｏｎ方法，用牛的新鲜坐骨神经制备髓鞘碱性蛋白（ＭＢＰ），并对所提取的ＭＢＰ进行生化及生物活性鉴定，测得ＭＢＰ的分子量为１４４００～２００００道尔顿之间，用所提取的ＭＢＰ免疫新西兰白兔，观察临床表现并行病理检查，１０只兔子有７只出现不同程度的临床症状，发病率达７０％，病理检查发现，发病兔炎细胞浸润以颈神经根、腰神经根、马尾神经损害重于坐骨神经干（Ｐ＜０．０１），单神经纤维分离，半薄切片及电镜下均发现以脱髓鞘改变为主，少数重症动物有轴索变性，脑脊膜有轻度充血及少量炎细胞浸润，脑实质内未见病变。     

实验性变态反应性神经炎动物模型的建立和病理特点

参照Ｙ．Ｌｏｎｄｏｎ方法，用牛的新鲜坐骨神经制备髓鞘碱性蛋白（ＭＢＰ），并对所提取的ＭＢＰ进行生化及生物活性鉴定，测得ＭＢＰ的分子量为１４４００～２００００道尔顿之间，用所提取的ＭＢＰ免疫新西兰白兔，观察临床表现并行病理检查，１０只兔子有７只出现不同程度的临床症状，发病率达７０％，病理检查发现，发病兔炎细胞浸润以颈神经根、腰神经根、马尾神经损害重于坐骨神经干（Ｐ＜０．０１），单神经纤维分离，半薄切片及电镜下均发现以脱髓鞘改变为主，少数重症动物有轴索变性，脑脊膜有轻度充血及少量炎细胞浸润，脑实质内未见病变。     

地塞米松对实验性变态反应性脑脊髓炎小鼠胸腺细胞凋亡的影响

目的 探讨地塞米松对实验性变态反应性脑脊髓炎（ＥＡＥ）小鼠胸腺细胞凋亡的影响。方法 采用昆明小鼠建立ＥＡＥ模型,于处死前２４小时腹腔注射米松,通过荧光染色法、原位末端标记法和电镜法观察地塞米松处理组,自然病程组和对照组的小鼠胸腺细胞凋亡。结果 地塞米松处理组的胸腺细胞凋亡率明显高于自然病程组和正常对照组（Ｐ〈０．０１）;且电镜发现有典型的凋亡改变。结论 地塞米松可明显增强ＥＡＥ小鼠胸腺细胞凋亡。     

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。     

硫脂致敏的实验性变态反应性神经炎病理和电生理特点

目的探讨硫脂诱导的实验性变态反应性神经炎病理和电生理特点。方法以硫脂免疫豚鼠,观察其周围神经电生理、病理及免疫细胞化学的变化。结果被致敏动物坐骨神经、脊神经根发生脱髓鞘改变,神经鞘膜ＩｇＧ荧光沉积明显,有的同时伴有髓鞘再生,病损区ＣＤ１１ｂ＋细胞显著增加而ＣＤ５＋细胞增加不明显,坐骨神经传导阻滞及传导速度减慢。结论硫脂致敏的实验性变态反应性神经炎与慢性格林－巴利综合征更相似。     

实验性变态反应性神经炎T细胞受体Vβ基因的表达和特征

目的 探讨在实验性变态反应性神经炎(EAN)中特异性识别抗原并呈优势扩增的为哪些Vβ亚家族基因.方法 应用逆转录-聚合酶链反应(RT-PCR)、原位杂交技术,比较T细胞受体(TCR)Vβ2、6、8、10、14基因mRNA在EAN组和对照组大鼠外周血、淋巴结及周围神经组织中的表达水平.结果 (1) TCRVβ6、8基因mRNA在EAN大鼠淋巴结中的表达(个/HP,下同)早期即有升高(分别为41.1±1.1和74.4±1.9,对照组为25.9±1.5和26.1±1.6),至发病高峰期更加明显,恢复期则与对照组无显著差异.(2) EAN组大鼠周围神经浸润T淋巴细胞的TCRVβ 6、8基因mRNA的表达从潜伏期(分别为4.9±2.9和11.7±9.1)至发病高峰期(分别为18.8±5.8和36.5±6.3)逐步增加,恢复期又有所下降,差异有极显著意义.(3) EAN大鼠发病早期及高峰期淋巴结与周围神经浸润淋巴细胞中TCRVβ 8基因mRNA的表达高于TCRVβ 6基因,差异有极显著意义.结论 T细胞上识别和限制EAN发病的特异性抗原的TCRVβ基因亚家族为TCRVβ 6和Vβ 8,以Vβ 8为主;特异性表达TCRVβ 6、8基因的T淋巴细胞在淋巴结中被激活并克隆性扩增,继而迁移至病变的周围神经组织中,可能导致一系列的免疫损害.     

Suppression of experimental allergic neuritis by cyclosporin-A

Summary Experimental allergic neuritis (EAN) was induced in guinea pigs and rats and treated with Cyclosporin-A (Cy-A). When Cy-A was given prophylactically for 1 month from the time of induction of the disease, it prevented the development of EAN during the course of its administration. When Cy-A was given therapeutically after the onset of neurological signs, it effectively prevented further deterioration. This effect was more marked after 3 weeks' treatment than after only 1 week's treatment.In both regimens, when dosing with Cy-A ceased there was a latent period before clinical signs of EAN developed. This latent period is similar to that seen in the development of EAN in normal control animals and is probably due to the continued presence of antigen at the injection sites. After primary treatment of EAN with Cy-A, animals that relapsed did not respond to further treatment with Cy-A.Histological examination revealed that the nature of the EAN lesions in both groups of animals given Cy-A were not as severe as those seen in control animals. Despite these observations, there was no statistically significant difference between the maximum clinical grades reached by animals in any one group.These experiments suggest that T-cells are important in the development of EAN and that Cy-A interferes with this process by suppressing T-helper cells. They also show that it is possible to influence favourably the course of immune mediated neurological disease.Presented in part at the 1st International Congress of Neuroimmunology, Stresa, Italy, 27th September–1st October, 1982     

Early myelin lesions in experimental allergic neuritis

We examined the evolution of demyelination in spinal roots of Lewis rats immunized with whole nerve and complete Freund' adjuvant. Roots were morphologically normal until 11 days after immunization, when we found endoneurial edema and myelin vesiculation in the absence of mononuclear cell contacts. Macrophage-associated myelin stripping was not detected until day 12. Macrophage infiltrations were extensive by day 14, but lymphocytes were sparse. These observations indicate that in experimental allergic neuritis, myelin injury may occur before macrophage-mediated demyelination, and provide support for an early role of serum factors in the development of this disorder.     

Autonomic dysfunction in experimental allergic neuritis

Beat-to-beat variation (R-R variation) in the electrocardiogram was studied in experimental allergic neuritis in the Sprague-Dawley rat. Reduced R-R variations were found in 2 of 10 animals, probably as a sign of autonomic dysfunction. The vagal nerves from these two animals, studied in vitro, showed disturbed conduction. In one animal prolonged conduction latencies to supramaximal electrical stimuli were found. Vagal nerves from controls and from animals without clinical symptoms showed normal conduction. Histologically, the vagal nerves from affected animals showed a slight inflammatory cell infiltration and signs of demyelination but there was no evidence of involvement of the brainstem vasomotor nuclei. Thus, we suggest that the autonomic dysfunction in experimental allergic neuritis, measured as reduced R-R variations, is caused by a peripheral vagal neuropathy.     

Axonal regeneration in experimental allergic peripheral neuritis

Summary It has recently been suggested that severed axons fail to regenerate in the mammalian central nervous system as a result of an autoimmune reaction to myelin basic protein released into the circulation at the time of injury. Since the autoantigenic components of peripheral myelin are rapidly phagocytosed after axonal transection, it is claimed that a comparable immune response does not occur following injury to peripheral nerves, so the regenerative process is not hindered. If this contention is correct, it should be possible to inhibit the regeneration of peripheral axons by inoculating animals with suitable neuritogenic homogenates of peripheral nervous tissue.It has been shown that axonal regeneration proceeds at the same rate in rats with experimental allergic neuritis as in healthy controls inoculated only with Freund's adjuvant. It is unlikely, therefore that myelin basic proteins can stimulate the production of antibodies capable of inhibiting regenerative axonal growth.     

In situ demonstration of T cell activation and elimination in the peripheral nervous system during experimental autoimmune neuritis in the Lewis rat

The mechanisms of activation and termination of autoimmune responses are poorly understood. We have studied the sites and mode of activation and elimination of T cells in actively induced experimental autoimmune neuritis (EAN) and in EAN adoptively transferred by P2-specific T cells (AT-EAN). The bromodeoxyuridine (BrdU) technique was employed to detect in situ proliferating cells in spleen and sciatic nerve. We assessed the nuclear morphology of infiltrating T cells using morphological criteria of apoptotic cell death. Apoptosis of lymphoid cells was also investigated using molecular labeling techniques. In AT-EAN, the number of BrdU-positive cells in splenic germinal centers peaked at day 2 after cell transfer [554 ± 267 (mean ± SEM) per mm²; controls 98 ± 35), 1 day before disease onset, and declined thereafter. BrdU incorporation in spleens from animals with active EAN peaked at day 11, around disease onset, but reached lower total values (165 ± 29 per mm²). In neither model did we observe a significant proportion of BrdU-positive T cells in the peripheral nervous system. However, T cells exhibiting morphological signs of apoptosis were detected in the sciatic nerve immediately after disease onset. The number of these cells was highest on day 7 in AT-EAN (6.6 ± 3.2 per mm²) and on day 17 in active EAN (11.2 ± 2.2 per mm²), corresponding to the maximum of T cell infiltration in both animal models. T cell activation occurs systemically and not just in the autoimmune lesion. Infiltrating T cells are eliminated by apoptosis in situ, terminating the inflammatory process. Further insight into these mechanisms may help to develop new therapeutic strategies for autoimmune disorders of the peripheral nervous system. Received: 21 July 1995 / Revised, accepted: 31 October 1995     

Detection of immune complexes in experimental allergic neuritis

Circulating immune complexes were assayed in the sera of animals with experimental allergic neuritis (EAN), and immunofluorescent staining and immunohistological examination were performed to clarify the role of immune complexes in the pathogenesis of EAN. The level of immune complexes in the sera of animals with clinical signs of EAN was from 1:32 to 1:64. Control animals showed a titer of immune complexes from 1:2 to 1:4. Animals with EAN showed deposition of immune complexes, which were detected by FITC-conjugated anti-rat IgG or C3-complement, in the vessels of the peripheral nerve. These findings suggest that immune complexes may contribute to the immunopathogenesis of EAN.     

The treatment of experimental allergic neuritis by plasma exchange

Experimental allergic neuritis (EAN) is a demyelinating disease of the peripheral nervous system that can be induced in laboratory animals. This disorder has been considered to show many similarities to acute inflammatory polyneuropathy (Guillain-Barré syndrome, GBS). Reports that plasma exchange may benefit patients with GBS prompted the investigation of the effect of plasma exchange in EAN. A controlled study was performed on New Zealand White rabbits. Sixteen animals were allocated to control or treatment groups at the onset of the disease. Clinical assessment on days 7 and 14 showed that treated animals were less severely affected neurologically (P = 0.05, day 7; P < 0.001 day 14), with a commensurate reduction in the severity of the histological lesions in peripheral nerves.     

Circulating lymphocyte subpopulations in experimental allergic neuritis

T-cell subsets in the peripheral blood were analyzed using monoclonal antibodies during the development of experimental allergic neuritis in Lewis rats. Percentages of helper and suppressor cells and ratios of helper/suppressor cells did not exceed normal limits during the development of the disease.     

Complement depletion affects demyelination and inflammation in experimental allergic neuritis

The effect of systemic complement depletion by cobra venom factor (CVF) on experimental allergic neuritis (BAN) was studied in rats immunized with variable amounts of bovine peripheral nerve myelin. Low-dose myelin EAN rats treated with CVF i.p. (n = 10) had lower clinical scores (0.3 ± 0.7 vs. 1.1 ± 1.1), less demyelination (0.4 ± 0.8 vs. 1.9 ± 1.1) and inflammation (0.6 ± 1.2 vs. 2 ± 1) than EAN animals treated with i.p. saline (n = 10). Endoneurial infiltrates had fewer EDI-positive (phagocytic) macrophages (if0.4 ± 0.5 vs.> 1.6 ± 1.1) and CD11bc-positive (expressing iC3b receptor or CR3) cells (1 ± 0.8 vs. 2.5 ± 0.8) (mean ± S.D.) detected by immunocytochemistry. This effect was partially abrogated by immunizing animals with a higher dose of myelin. Our studies suggest that complement may play a role in the recruitment of macrophages into the endoneurium and in opsonizing myelin for phagocytosis.