脱氢表雄酮对实验性自身免疫性神经炎保护作用的研究
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摘要
研究目的
格林-巴利综合征(Guillain-Barre syndrome,GBS)是引起迟缓性麻痹的主要疾病,是外周神经的炎症性脱髓鞘疾病。实验性自身免疫性神经炎(experimental autoimmune neuritis,EAN)是CD4~+ T细胞介导的外周神经自身免疫性脱髓鞘疾病,其临床经过、神经电生理检查及病理学改变与GBS最常见的临床亚型-急性炎症性脱髓鞘性多发神经炎(acute inflammation demyelinative polyradiculoneuropathy,AIDP)极为相似,是公认的研究GBS的经典动物模型。目前,GBS的治疗主要是血浆置换或静脉注射丙种球蛋白,虽然其能缩短病程,加快病情恢复,但仍有大约5%病死率,20%的患者遗留永久性残疾。多数学者认为,糖皮质激素等免疫抑制剂的对本病无效。因此,有必要探讨治疗GBS的新方法。
脱氢表雄酮(dehydroepiandrosterone,DHEA)是一种19碳类固醇,是雌激素和雄激素的前体物质,也是目前尚不明确的多种免疫调节性类固醇的前体物质。体内、外研究证实DHEA及其代谢产物具有抗炎、抗增殖和免疫调节活性,对人和动物的免疫系统可产生有益的作用。EAN是Th1细胞介导的外周神经自身免疫性脱髓鞘疾病,神经内膜下T淋巴细胞、巨噬细胞浸润及多灶性、节段性髓鞘脱失为主要病理改变,促炎性细胞因子(如IFN-γ、TNF-α、IL-12、NO等)在其发病机制中起关键作用。目前国内外尚未见有应用DHEA治疗EAN的报道,基于以往体内外研究证实的DHEA对免疫系统的作用,我们应用DHEA治疗发病初期EAN,通过观察EAN的临床、病理及免疫指标,评价DHEA对发病初期EAN的治疗作用并探讨其免疫保护机制。
方法
1.抗原:采用超速离心法自牛脊神经根提取周围神经髓磷脂(bovine peripheral nerve myelin,BPM)。
2.实验动物:6-8周龄雌性Lewis大鼠36只,体重150-170g,SPF级饲养。
3.EAN模型的建立及临床评分:大鼠经适应性饲养一周,氯胺酮(10mg/Kg)麻醉。将100μl抗原乳剂(含BPM 5mg、结核杆菌1mg、不完全弗氏佐剂50μl、生理盐水50μl)注入双侧后肢足垫皮下。于免疫前即时及免疫后每日进行临床评估,直至免疫后28天。临床评分:0:正常;0.5:全尾肌张力下降;1.0:尾部肌肉瘫,松软拖地;1.5:轻微后肢无力,肌张力下降;2.0:明显后肢无力或轻度后肢瘫;2.5:中度后肢瘫;3.0:严重后肢瘫;4:严重四肢瘫。
4.脱氢表雄酮(DHEA)治疗:将大鼠随机分为DHEA 0.5mg/剂治疗组、2mg/剂治疗组和对照组,每组各12只。治疗组于免疫后第5天开始皮下注射DHEA,体积50μl,每日一次,至免疫后28天。对照组皮下注射相同体积的DHEA溶媒二甲基亚砜(DMSO)。
5.组织病理学检查:于免疫后16天处死大鼠,每组各7只,取坐骨神经近脊髓段,10%福尔马林固定,石蜡包埋,纵行切片(5-6μm,每根神经2张连续切片),HE染色,应用计算机图像分析系统40倍物镜下采集图像,每张切片随机取5个视野,计数坐骨神经中浸润的单个核细胞,取其平均值,结果以细胞数/mm~2组织表示。
6.免疫组化检测:大鼠坐骨神经石蜡切片,常规脱蜡致水,经阻断内源性过氧化物酶活性、抗原修复、牛血清白蛋白封闭处理,分别滴加一抗(山羊抗大鼠IFN-γ、TNF-α多克隆抗体)和二抗(辣根过氧化酶标记兔抗山羊IgG),DAB显色,苏木苏复染,脱水,透明,封固。40倍物镜下采集图像,观察大鼠坐骨神经中细胞阳性反应,结果以细胞数/mm~2组织表示。
7.单个核细胞体外增殖试验:取免疫后16天EAN大鼠腹股沟淋巴结和脾脏制备单个细胞悬液(2×10~6/ml),悬浮于RPMI1640培养基,接种到96孔培养板,分别加入10μl BPM、植物血凝素(phytohemagglutinin,PHA)和磷酸盐缓冲溶液(phosphate bufferedsolution,PBS)培养48小时,再加入~3H-甲基胸腺嘧啶脱氧核苷1μCi/孔,18小时后收集细胞,β-液体闪烁仪检测测定胸腺嘧啶核苷结合量,结果以每分钟放射性荧光闪烁计数值(cpm)表示。
8.单个核细胞培养上清液细胞因子检测:来自引流淋巴结和脾脏的单个核细胞悬液(2×10~6/ml),加入BPM(终浓度20μg/ml),培养48小时,收获上清液。应用酶联免疫分析试剂盒检测TNF-α、IFN-γ、IL-10,操作按说明书步骤进行。标准品和样本均设双复孔,结果以pg/ml表示。
9.统计学方法:应用SPSS 13.0软件对资料进行统计分析,数据以(?)±s表示。应用单因素方差分析或秩和检验(Kruskal-Wallis检验法)进行组间比较,组间差异有统计学意义则用SNK法或Bonferroni法进行组间两两比较,P＜0.05为差别有统计学意义。
结果
1.DHEA延迟EAN发病时间:三组大鼠在发病时间上表现出明显差异,DHEA 2mg治疗组发病时间为11.83±0.94天,0.5mg治疗组为10.92±1.08天,对照组为9.17±0.94天。与对照组比较,两种剂量DHEA治疗组发病时间均延迟(P＜0.05,P＜0.05),DHEA 2mg治疗组与0.5mg治疗组比较发病时间亦明显延迟(P＜0.05)。
2.DHEA降低EAN高峰期临床评分:疾病高峰期(免疫后第16d)临床评分显示DHEA 2mg治疗组为1.42±0.73,0.5mg治疗组为2.00±0.56,对照组为2.29±0.62。与对照组比较,DHEA 2mg治疗组疾病高峰期临床评分明显降低(P＜0.05)。DHEA 2mg治疗组疾病高峰期临床评分明显低于0.5mg治疗组(P＜0.05)。0.5mg治疗组疾病高峰期评分低于对照组,但经统计学处理无显著差异。
3.DHEA减少炎性细胞在外周神经浸润:疾病高峰期大鼠坐骨神经组织病理学检查结果显示,DHEA 2mg治疗组骨神经中炎性细胞浸润数目为37.00±29.59个/mm~2组织,0.5mg治疗组为62.00±37.11/mm~2组织,对照组为105.50±31.05/mm~2组织。与对照组比较,DHEA 2mg治疗组和0.5mg治疗组坐骨神经中炎性细胞浸润数目均明显减少(P＜0.05,P＜0.05)。DHEA 2mg治疗组炎性细胞浸润数目较0.5mg治疗组减少,但经统计学处理无意义。
4.DHEA抑制脾脏单个核细胞增殖反应:于EAN高峰期高峰期检测大鼠引流淋巴结和脾脏单个核细胞体外增殖,BPM、PHA诱导的和无抗原刺激(PBS)组单个核细胞增殖(cpm)分别为:DHEA 2mg治疗组2900±672、4500±1581和594±220;0.5mg治疗组5125±1433、6562±1720和1150±160;对照组9125±1941、11680±2344和1338±358。结果显示,与对照组比较两种剂量DHEA治疗组均能明显抑制BPM和PHA诱导的单个核细胞增殖(P＜0.05,P＜0.05,P＜0.05,P＜0.05);与对照组比较,DHEA 2mg治疗组对无抗原刺激组(PBS)单个核细胞增殖有明显抑制作用(P＜0.05),0.5mg治疗组与对照组比较差异无统计学意义。
5.DHEA减少外周神经促炎细胞因子产生:应用免疫组化技术检测疾病高峰期大鼠坐骨神经中IFN-γ、TNF-α表达,结果显示IFN-γ、TNF-α阳性反应细胞数分别为:DHEA 2mg治疗组17.50±13.03个/mm~2组织、13.63±10.03个/mm~2组织,0.5mg治疗组25.75±13.78个/mm~2组织、20.50±11.41个/mm~2组织,对照组64.88±23.22个/mm~2组织、47.00±16.63个/mm~2组织。与对照组比较,DHEA 2mg治疗组和0.5mg治疗组IFN-γ、TNF-α阳性反应细胞数均明显减少(P＜0.05,P＜0.05,P＜0.05,P＜0.05),DHEA 2mg治疗组与0.5mg治疗组比较IFN-γ、TNF-α阳性反应细胞数未见明显差异。
6.DHEA影响单个核细胞培养上清液细胞因子水平:应用酶联免疫吸附法检测引流淋巴结和脾脏单个核细胞培养上清中IFN-γ、TNF-α、IL-10水平,结果显示DHEA 2mg治疗组分别为143.13±43.17 pg/ml、90.00±29.76 pg/ml、70.13±26.22 pg/ml;0.5mg治疗组分别为213.75±44.06 pg/ml、119.38±19.35 pg/ml、46.00±12.62 pg/ml,对照组为365.00±63.70 pg/ml、176.25±28.75 pg/ml、48.36±16.34pg/ml。与对照组比较,DHEA 2 mg治疗组和0.5 mg治疗组均能减少IFN-γ和TNF-α生成(P＜0.05,P＜0.05,P＜0.05,P＜0.05),DHEA2 mg治疗组IFN-γ和TNF-α水平较0.5 mg治疗组降低(P＜0.05,P＜0.05);各组间IL-10水平未显示明显的差异。
结论
1.应用两种剂量脱氢表雄酮(DHEA)治疗发病初期EAN,与对照组比较,可明显延缓发病时间,降低疾病高峰期临床评分,减少坐骨神经中炎性细胞浸润数目,提示DHEA对外周神经自身免疫性疾病有治疗作用。
2.应用两种剂量DHEA治疗发病初期EAN,与对照组比较,可明显抑制BPM诱导的单个核细胞增殖,减少坐骨神经中IFN-γ、TNF-α阳性反应细胞数目,降低BPM刺激的单个核细胞培养上清中IFN-γ、TNF-α水平。提示DHEA可抑制EAN大鼠自身反应性T淋巴细胞增殖和促炎性细胞因子过度表达,抑制细胞免疫反应。
3.DHEA可降低全身和病灶局部IFN-γ、TNF-α水平,对IL-10产生无影响。表明DHEA对EAN的治疗作用主要是下调Thl1反应,抑制促炎性细胞因子过度分泌,对Th2细胞因子水平无影响。
4.与0.5mg/剂治疗组比较,2mg/剂治疗组能明显延缓EAN发病时间,降低疾病高峰期临床评分,减少单个核细胞培养上清IFN-γ、TNF-α产生,提示DHEA疗效的剂量相关性。
研究意义
格林-巴利综合征(GBS)引起急性迟缓性麻痹最常见的原因,是外周神经的自身免疫性炎症性脱髓鞘疾病。目前,血浆置换或静脉注射丙种球蛋白是治疗GBS的主要方法,其能缩短病程,加快恢复,但临床上仍有大约5%的病死率,大约20%的患者遗留永久性残疾。因此,有必要研究尝试新的治疗方法,阻断炎症级联反应,降低本病的死亡率及神经系统后遗症的发生率。
脱氢表雄酮(DHEA)是人类循环中最丰富的肾上腺皮质类固醇,体内、外研究证实DHEA及其代谢产物具有抗炎、抗增殖和免疫调节活性,对人和动物的免疫系统可产生有益的作用。实验性自身免疫性神经炎(EAN)是GBS最常见的临床亚型急性炎症性脱髓鞘性多发神经炎(AIDP)的经典动物模型,本课题应用DHEA治疗发病初期EAN,通过观察EAN的临床和病理及免疫指标的改变,评价DHEA对发病初期EAN的治疗作用并探讨其免疫机制,为探索治疗自身免疫性疾病的新模式提供了依据。
创新性
1.本课题应用牛周围神经髓磷脂与完全弗氏佐剂混合制成抗原乳剂,注入易感鼠种Lewis大鼠双后肢足垫皮下,诱导实验性自身免疫性神经炎(EAN)动物模型。该模型是GBS最常见的临床亚型AIDP的经典动物模型,目前国内未见有应用上述方法及鼠种诱导EAN的报道。
2.本课题应用脱氢表雄酮(DHEA)治疗发病初期EAN,观察比较临床评分、坐骨神经炎性细胞浸润,评价DHEA的治疗效果。通过观察DHEA对引流淋巴结和脾脏单个核细胞体外增殖、培养上清IFN-γ、TNF-α、IL-10水平及坐骨神经中IFN-γ、TNF-α表达水平的影响,探讨其免疫治疗机制。目前国内外尚未见DHEA治疗EAN及其机制的研究报道。
Objective
Guillain-Barre syndrome(GBS) is the main cause of crippling disease in humans and characterized as acute demyelinating inflammation in structures of the peripheral nervous system(PNS).Experimental autoimmune neuritis(EAN) is a T cell-mediated,acute,autoimmune demyelination disease of the PNS that serves as a model for human GBS. The close clinical,histopathological,and electrophysiological similarities between EAN and GBS make EAN a classic animal model for studying pathogenic mechanisms and novel treating strategies for GBS. For the treatment of GBS,both plasma exchange and intravenous immunoglobulin have shown the efficacy in shorten the progressive course.But there is a mortality of approximately 5%,and approximately 20%of patients are left with significant and persistent disability.Many of the conventional immunosuppressive treatments,such as corticosteroids, have no effect on GBS.It is needed to uncover new therapeutic modalities for GBS.
Dehydroepiandrosterone(DHEA),a C19 adrenal steroid,is a precursor of both androgens and oestrogens.DHEA also serves as a precursor for many less well understood immune-regulating steroids. Studies showed that DHEA and its metabolites have anti-inflammatory, anti-proliferative,and certain immune-regulating properties both in vitro and in vivo.DHEA have shown beneficial effects on immunologic system in experimental animals and humans.EAN is a Thl-mediated autoimmune demyelination disease of the PNS.The infiltration of T cells and macrophages in the PNS and the multifocal,segmental demyelination are the main pathological changes of EAN,in which inflammatory cytokines, such as IFN-γ、TNF-α、IL-12、NO,play important roles.Thus far,there is no report about the role of DHEA in EAN.Based on the previous findings of DHEA on immunologic system,DHEA was used in treatment of EAN in the initial phase in this study.The therapeutical effect of DHEA on EAN and the immunologic mechanism were evaluated by examining the changes in clinical,histopathological and immunological parameters. Methods
1.Antigens:Bovine peripheral nerve myelin(BPM) was prepared from intra- and extradural spinal nerve roots by ultracentrifugation.
2.Animals:Thirty-six female Lewis rats,6-8 weeks old,weighing 150 - 180g were used in the present study.All of the rats were maintained in specific- pathogen free condition.
3.Induction and clinical evaluation of EAN:Ketamine was used for the animal anesthesia(10mg/Kg).EAN was induced by subcutaneous(s.c.) injection into both hind footpads with 100μl of inoculum containing 5 mg of BPM and 2 mg of Mycobacterium tuberculosis and 50μl Freund's incomplete adjuvant.Clinical scores were assessed immediately before immunization and thereafter every day until day 28 post immunization (p.i.).Severity of paresis was graded as follows:0= no illness;1= flaccid tail;2= dragging both hind limbs;3= paralysis of both hind limbs;4= paralysis of four limbs or death;intermediate scores of 0.5 increment were given to rats with intermediate signs.
4.DHEA treatment:Thirty-six Lewis rats were randomly divided into DHEA 0.5mg treatment group,2mg treatment group and control group(n = 12).Treatment groups were subcutaneously injected every day with DHEA and control group were subcutaneously injected with the same volum of DHEA dissolvent DMSO from day 5 p.i.to day 28 p.i.
5.Histopathological assessment:Seven rats from each group were killed on day 16 p.i.,at a time when the clinical signs of EAN peaked. Sciatic nerve segments were excised close to the lumbar spinal cord, fixed in 10%phosphate-buffered formalin,and embedded in paraffin. Multiple longitudinal sections(5- 6μm slices) of sciatic nerves were stained with hematoxylin and eosin for evaluation of the extent of mononuclear cell(MNC) infiltration by light microscopy.Tissue areas were measured by image analysis and the numbers of inflammatory cells were counted at×40 magnification.The average results were expressed as cells per mm~2 tissue section.
6.Immunohistochemistry:Paraffin tissue sections(5μm) were deparaffinized,hydrated and treated for blocking endogenous peroxidase activity,repairing antigen.The sections were incubated with polyclonal goat anti-rat IFN-γ、TNF-αantibody for 1h and then stained according to the avidin-biotin technique.The numbers of positive cells were counted at×40 magnification in the entire section area.The average results were expressed as cells per mm~2 tissue section.
7.Lymphocyte proliferation assay:The draining lymph nodes and spleens were removed under aseptic conditions.Single cell suspensions of MNC from individual rats were prepared separately.The cells were suspended in RPMI1640 and cultured in triplicates in round-bottomed 96-well microtitre plates in the presence of 10μl of BPM or phytohemagglutinin or the same volume of PBS.After 48h of incubation, proliferation was measured by labeling cells with[~3H]-methylthymidine (1μCi/well) and cultured for an additional 18h.Cells were harvested onto glass fibre filters.[~3H]-methylthymidine incorporation was measured in liquidβ- scintillation counter.Data are given as mean counts per minute (cpm).
8.Measurement of cytokine:MNC were cultured at a cell density of 2×10~6 cells/ml in medium containing BPM(20μg/ml).Supernatants were collected after 48h of incubation at 37℃.Quantitative ELISA assays for cytokines of IFN-γ,TNF-αand IL-10 were performed using ELISA kits following the manufacturer's instructions.Determinations were performed in duplicate and results were expressed as pg/ml.
9.Statistical analysis:The SPSS 13.0 computer program was used for all calculations and statistical evaluations.Results were expressed as means±standard deviation(SD).Differences between groups were evaluated by one-factor analysis of variance(ANOVA) or rank-sum test. The level of significance was set to p＜0.05.
Results
1.DHEA treatment delayed disease onset of EAN:Both DHEA-treated groups exhibited more delayed disease onset than control rats(P＜0.05, P＜0.05).Compared to 0.5 mg DHEA treatment group,2 mg group also displayed significant delayed disease onset(P＜0.05).
2.DHEA treatment decreased disease peak clinical scores of EAN: The rats treated with DHEA at dose of 2 mg/day showed significantly lower mean peak clinical scores than the control rats(P＜0.05).0.5 mg DHEA treatment group showed lower mean peak clinical scores than the control group,but there was no statistically significant difference.
2.DHEA treatment inhibits inflammatory cell infiltration of PNS:The sciatic nerves of EAN rats were examined histologically at the height of the clinical course of EAN.The results revealed that administration of DHEA at doses of both 2 mg/day and 0.5 mg/day all significantly reduced the infiltrating cells in sciatic nerves sections compared to control EAN rats(P＜0.05,P＜0.05).2 mg DHEA treatment group showed less numbers of infiltrating cells in sciatic nerves than 0.5 mg treatment group,but there was no significant difference statistically.
3.Effects of DHEA on MNCs Proliferation:MNCs of draining lymph nodes and spleens from EAN rats were tested on day 16 p.i.for lymphocytes proliferation.When compared to the controls,the DHEA treated rats showed significantly suppressed BPM and PHA-induced MNC proliferation(P＜0.05,P＜0.05,P＜0.05,P＜0.05 ).In addition,there was also a lower level of proliferation in 2 mg DHEA treatment rats than control EAN rats when MNC were cultured in the absence of antigen(P＜0.05),but there was no statistically significant difference between 0.5 mg DHEA treatment group and the controls.
4.DHEA treatment suppresses the cytokine expression in sciatic nerves:Immunohistochemical studies showed that the numbers of IFN-γand TNF-α.expressing cells were strongly decreased in the sciatic nerves of DHEA treated rats at dose of 2 mg/day and 0.5 mg/day compared to control EAN rats(P＜0.05,P＜0.05,P＜0.05,P＜0.05 ).No statistically confirmed difference regarding the numbers of cells expressing IFN-γ,and TNF-αin the PNS was shown when comparing 2 mg-DHEA to 0.5 mg -DHEA treatment groups.
5.Levels of cytokine in supernatant:The levels of IFN-γ,TNF-αand IL-10 secretion from MNC supernatants were examined by ELISA.The results showed that levels of IFN-γ,and TNF-α.in the MNC supernatant were significantly decreased in EAN rats treated with DHEA at doses of 2 mg/day and 0.5 mg/day when examined on day 16 p.i.as compared with control EAN rats(P＜0.05,P＜0.05,P＜0.05,P＜0.05).There were also statistically significant differences in IFN-γ,and TNF-αsecretion in the MNC supernatants between 2 mg DHEA and 0.5 mg DHEA treated EAN rats(P＜0.05,P＜0.05).There were no differences in IL-10 production among the three groups of rats.
Conclusion
1.In this study,DHEA was administered to EAN rats at doses of 0.5 mg/rat/day and 2 mg/rat/day for the treatment of EAN in the initial phase. Rats treated with DHEA at the both doses displayed significant delay in onset,lower peak clinical score and decreased inflammatory cell infiltration in the PNS as compared to control rats.These data suggest that DHEA has therapeutic potential for alleviating EAN.
2.Rats treated with DHEA at the both doses displayed significant decreases in numbers of IFN-γ,and TNF-αexpressing cells in the PNS, BPM-stimulated MNCs proliferation and IFN-γ,TNF-α-secretion in the draining lymph node and spleen cells.Our finding suggests that DHEA could inhibit cellular immunological response by suppressing the proliferation of autoreactive T cells and overexpression of proinflamation cytokines.
3.In the present study,administration of DHEA to EAN rats decreased IFN-γ、TNF-αsecretion both systemically and within sites of inflammation,with no significant changes in IL-10 production.Our finding suggests that DHEA suppressed EAN mainly by downregulation of autoimmune Th1 response,decreasing the production of proinflammatory cytokines without changing levels of Th2 associated cytokines.
4.Our data show that rats treated with DHEA at dose of 2mg every day exhibited delayed disease onset,lower mean peak clinical scores and decreased levels of IFN-γ,and TNF-α.from cultured MNC supernatants when compared to 0.Smg DHEA-treated EAN rats.The results suggest that the immune suppressive effect of DHEA is dose-dependent.
Significance
Guillain-Barre syndrome(GBS) is the most main cause of crippling disease in humans and characterized as acute demyelinating inflammation in structures of the peripheral nervous system.For the treatment of GBS, both plasma exchange and intravenous immunoglobulin have shown the efficacy in shorten the progressive course and hastening recovery from GBS.But there is a mortality of approximately 5%,and approximately 20%of patients are left with significant and persistent disability.Further research is needed to uncover new therapeutic modalities for GBS to block the cascade inflammation and decreas the mortality and sequelae of nervous system.
Dehydroepiandrosterone(DHEA) is the most abundant circulating adrenal steroids in humans.It has been well-documented that DHEA and its metabolites have anti-inflammatory,anti-proliferative,and certain immune-regulating properties and have shown immunological effects both in vitro and in vivo in experimental animals and humans.Experimental autoimmune neuritis(EAN) serves as classic animal model for acute inflammatory demyelinative polyradiculoneuropathy(AIDP) - one of the most common subtype of human.In this study,the therapeutical effect of DHEA on the initial phase of EAN and the immunologic mechanism were evaluated by examining the changes in clinical,histopathological and immunological parameters.We aimed to provide evidence for searching a promising new and potentially powerful strategy for human autoimmune disorders.
creativity
1.In our study,the inoculum was prepared by emulsifying bovine peripheral nerve myelin(BPM) in Freund's complete adjuvant(CFA). EAN was induced by subcutaneous injection the inoculum into both hind footpads of susceptible animals - Lewis rat.EAN serves as a classical model for human AIDP,which is the most common subtype of GBS.So far,there has been no report on inducing EAN with BPM and CFA in Lewis rat in China.
2.In this study,DHEA was used for the treatment of EAN in the initial phase.The therapeutic effect of DHEA was evaluated by assessing clinical scores and examining inflammatory cell infiltration in the PNS. The immunologic mechanism were evaluated by examining the changes in BPM-stimulated MNCs proliferation from draining lymph nodes and spleens,IFN-γ,TNF-β,IL-10 -secretion from MNC supernatants and IFN-γ,TNF-αexpressing cells in the PNS.Up to now,there has been no report about DHEA administration for the treatment of EAN at home and abroad.
格林-巴利综合征(Guillain-Barre syndrome,GBS)是引起迟缓性麻痹的主要疾病,是外周神经的炎症性脱髓鞘疾病。实验性自身免疫性神经炎(experimental autoimmune neuritis,EAN)是CD4~+ T细胞介导的外周神经自身免疫性脱髓鞘疾病,其临床经过、神经电生理检查及病理学改变与GBS最常见的临床亚型-急性炎症性脱髓鞘性多发神经炎(acute inflammation demyelinative polyradiculoneuropathy,AIDP)极为相似,是公认的研究GBS的经典动物模型。目前,GBS的治疗主要是血浆置换或静脉注射丙种球蛋白,虽然其能缩短病程,加快病情恢复,但仍有大约5%病死率,20%的患者遗留永久性残疾。多数学者认为,糖皮质激素等免疫抑制剂的对本病无效。因此,有必要探讨治疗GBS的新方法。
脱氢表雄酮(dehydroepiandrosterone,DHEA)是一种19碳类固醇,是雌激素和雄激素的前体物质,也是目前尚不明确的多种免疫调节性类固醇的前体物质。体内、外研究证实DHEA及其代谢产物具有抗炎、抗增殖和免疫调节活性,对人和动物的免疫系统可产生有益的作用。EAN是Th1细胞介导的外周神经自身免疫性脱髓鞘疾病,神经内膜下T淋巴细胞、巨噬细胞浸润及多灶性、节段性髓鞘脱失为主要病理改变,促炎性细胞因子(如IFN-γ、TNF-α、IL-12、NO等)在其发病机制中起关键作用。目前国内外尚未见有应用DHEA治疗EAN的报道,基于以往体内外研究证实的DHEA对免疫系统的作用,我们应用DHEA治疗发病初期EAN,通过观察EAN的临床、病理及免疫指标,评价DHEA对发病初期EAN的治疗作用并探讨其免疫保护机制。
方法
1.抗原:采用超速离心法自牛脊神经根提取周围神经髓磷脂(bovine peripheral nerve myelin,BPM)。
2.实验动物:6-8周龄雌性Lewis大鼠36只,体重150-170g,SPF级饲养。
3.EAN模型的建立及临床评分:大鼠经适应性饲养一周,氯胺酮(10mg/Kg)麻醉。将100μl抗原乳剂(含BPM 5mg、结核杆菌1mg、不完全弗氏佐剂50μl、生理盐水50μl)注入双侧后肢足垫皮下。于免疫前即时及免疫后每日进行临床评估,直至免疫后28天。临床评分:0:正常;0.5:全尾肌张力下降;1.0:尾部肌肉瘫,松软拖地;1.5:轻微后肢无力,肌张力下降;2.0:明显后肢无力或轻度后肢瘫;2.5:中度后肢瘫;3.0:严重后肢瘫;4:严重四肢瘫。
4.脱氢表雄酮(DHEA)治疗:将大鼠随机分为DHEA 0.5mg/剂治疗组、2mg/剂治疗组和对照组,每组各12只。治疗组于免疫后第5天开始皮下注射DHEA,体积50μl,每日一次,至免疫后28天。对照组皮下注射相同体积的DHEA溶媒二甲基亚砜(DMSO)。
5.组织病理学检查:于免疫后16天处死大鼠,每组各7只,取坐骨神经近脊髓段,10%福尔马林固定,石蜡包埋,纵行切片(5-6μm,每根神经2张连续切片),HE染色,应用计算机图像分析系统40倍物镜下采集图像,每张切片随机取5个视野,计数坐骨神经中浸润的单个核细胞,取其平均值,结果以细胞数/mm~2组织表示。
6.免疫组化检测:大鼠坐骨神经石蜡切片,常规脱蜡致水,经阻断内源性过氧化物酶活性、抗原修复、牛血清白蛋白封闭处理,分别滴加一抗(山羊抗大鼠IFN-γ、TNF-α多克隆抗体)和二抗(辣根过氧化酶标记兔抗山羊IgG),DAB显色,苏木苏复染,脱水,透明,封固。40倍物镜下采集图像,观察大鼠坐骨神经中细胞阳性反应,结果以细胞数/mm~2组织表示。
7.单个核细胞体外增殖试验:取免疫后16天EAN大鼠腹股沟淋巴结和脾脏制备单个细胞悬液(2×10~6/ml),悬浮于RPMI1640培养基,接种到96孔培养板,分别加入10μl BPM、植物血凝素(phytohemagglutinin,PHA)和磷酸盐缓冲溶液(phosphate bufferedsolution,PBS)培养48小时,再加入~3H-甲基胸腺嘧啶脱氧核苷1μCi/孔,18小时后收集细胞,β-液体闪烁仪检测测定胸腺嘧啶核苷结合量,结果以每分钟放射性荧光闪烁计数值(cpm)表示。
8.单个核细胞培养上清液细胞因子检测:来自引流淋巴结和脾脏的单个核细胞悬液(2×10~6/ml),加入BPM(终浓度20μg/ml),培养48小时,收获上清液。应用酶联免疫分析试剂盒检测TNF-α、IFN-γ、IL-10,操作按说明书步骤进行。标准品和样本均设双复孔,结果以pg/ml表示。
9.统计学方法:应用SPSS 13.0软件对资料进行统计分析,数据以(?)±s表示。应用单因素方差分析或秩和检验(Kruskal-Wallis检验法)进行组间比较,组间差异有统计学意义则用SNK法或Bonferroni法进行组间两两比较,P＜0.05为差别有统计学意义。
结果
1.DHEA延迟EAN发病时间:三组大鼠在发病时间上表现出明显差异,DHEA 2mg治疗组发病时间为11.83±0.94天,0.5mg治疗组为10.92±1.08天,对照组为9.17±0.94天。与对照组比较,两种剂量DHEA治疗组发病时间均延迟(P＜0.05,P＜0.05),DHEA 2mg治疗组与0.5mg治疗组比较发病时间亦明显延迟(P＜0.05)。
2.DHEA降低EAN高峰期临床评分:疾病高峰期(免疫后第16d)临床评分显示DHEA 2mg治疗组为1.42±0.73,0.5mg治疗组为2.00±0.56,对照组为2.29±0.62。与对照组比较,DHEA 2mg治疗组疾病高峰期临床评分明显降低(P＜0.05)。DHEA 2mg治疗组疾病高峰期临床评分明显低于0.5mg治疗组(P＜0.05)。0.5mg治疗组疾病高峰期评分低于对照组,但经统计学处理无显著差异。
3.DHEA减少炎性细胞在外周神经浸润:疾病高峰期大鼠坐骨神经组织病理学检查结果显示,DHEA 2mg治疗组骨神经中炎性细胞浸润数目为37.00±29.59个/mm~2组织,0.5mg治疗组为62.00±37.11/mm~2组织,对照组为105.50±31.05/mm~2组织。与对照组比较,DHEA 2mg治疗组和0.5mg治疗组坐骨神经中炎性细胞浸润数目均明显减少(P＜0.05,P＜0.05)。DHEA 2mg治疗组炎性细胞浸润数目较0.5mg治疗组减少,但经统计学处理无意义。
4.DHEA抑制脾脏单个核细胞增殖反应:于EAN高峰期高峰期检测大鼠引流淋巴结和脾脏单个核细胞体外增殖,BPM、PHA诱导的和无抗原刺激(PBS)组单个核细胞增殖(cpm)分别为:DHEA 2mg治疗组2900±672、4500±1581和594±220;0.5mg治疗组5125±1433、6562±1720和1150±160;对照组9125±1941、11680±2344和1338±358。结果显示,与对照组比较两种剂量DHEA治疗组均能明显抑制BPM和PHA诱导的单个核细胞增殖(P＜0.05,P＜0.05,P＜0.05,P＜0.05);与对照组比较,DHEA 2mg治疗组对无抗原刺激组(PBS)单个核细胞增殖有明显抑制作用(P＜0.05),0.5mg治疗组与对照组比较差异无统计学意义。
5.DHEA减少外周神经促炎细胞因子产生:应用免疫组化技术检测疾病高峰期大鼠坐骨神经中IFN-γ、TNF-α表达,结果显示IFN-γ、TNF-α阳性反应细胞数分别为:DHEA 2mg治疗组17.50±13.03个/mm~2组织、13.63±10.03个/mm~2组织,0.5mg治疗组25.75±13.78个/mm~2组织、20.50±11.41个/mm~2组织,对照组64.88±23.22个/mm~2组织、47.00±16.63个/mm~2组织。与对照组比较,DHEA 2mg治疗组和0.5mg治疗组IFN-γ、TNF-α阳性反应细胞数均明显减少(P＜0.05,P＜0.05,P＜0.05,P＜0.05),DHEA 2mg治疗组与0.5mg治疗组比较IFN-γ、TNF-α阳性反应细胞数未见明显差异。
6.DHEA影响单个核细胞培养上清液细胞因子水平:应用酶联免疫吸附法检测引流淋巴结和脾脏单个核细胞培养上清中IFN-γ、TNF-α、IL-10水平,结果显示DHEA 2mg治疗组分别为143.13±43.17 pg/ml、90.00±29.76 pg/ml、70.13±26.22 pg/ml;0.5mg治疗组分别为213.75±44.06 pg/ml、119.38±19.35 pg/ml、46.00±12.62 pg/ml,对照组为365.00±63.70 pg/ml、176.25±28.75 pg/ml、48.36±16.34pg/ml。与对照组比较,DHEA 2 mg治疗组和0.5 mg治疗组均能减少IFN-γ和TNF-α生成(P＜0.05,P＜0.05,P＜0.05,P＜0.05),DHEA2 mg治疗组IFN-γ和TNF-α水平较0.5 mg治疗组降低(P＜0.05,P＜0.05);各组间IL-10水平未显示明显的差异。
结论
1.应用两种剂量脱氢表雄酮(DHEA)治疗发病初期EAN,与对照组比较,可明显延缓发病时间,降低疾病高峰期临床评分,减少坐骨神经中炎性细胞浸润数目,提示DHEA对外周神经自身免疫性疾病有治疗作用。
2.应用两种剂量DHEA治疗发病初期EAN,与对照组比较,可明显抑制BPM诱导的单个核细胞增殖,减少坐骨神经中IFN-γ、TNF-α阳性反应细胞数目,降低BPM刺激的单个核细胞培养上清中IFN-γ、TNF-α水平。提示DHEA可抑制EAN大鼠自身反应性T淋巴细胞增殖和促炎性细胞因子过度表达,抑制细胞免疫反应。
3.DHEA可降低全身和病灶局部IFN-γ、TNF-α水平,对IL-10产生无影响。表明DHEA对EAN的治疗作用主要是下调Thl1反应,抑制促炎性细胞因子过度分泌,对Th2细胞因子水平无影响。
4.与0.5mg/剂治疗组比较,2mg/剂治疗组能明显延缓EAN发病时间,降低疾病高峰期临床评分,减少单个核细胞培养上清IFN-γ、TNF-α产生,提示DHEA疗效的剂量相关性。
研究意义
格林-巴利综合征(GBS)引起急性迟缓性麻痹最常见的原因,是外周神经的自身免疫性炎症性脱髓鞘疾病。目前,血浆置换或静脉注射丙种球蛋白是治疗GBS的主要方法,其能缩短病程,加快恢复,但临床上仍有大约5%的病死率,大约20%的患者遗留永久性残疾。因此,有必要研究尝试新的治疗方法,阻断炎症级联反应,降低本病的死亡率及神经系统后遗症的发生率。
脱氢表雄酮(DHEA)是人类循环中最丰富的肾上腺皮质类固醇,体内、外研究证实DHEA及其代谢产物具有抗炎、抗增殖和免疫调节活性,对人和动物的免疫系统可产生有益的作用。实验性自身免疫性神经炎(EAN)是GBS最常见的临床亚型急性炎症性脱髓鞘性多发神经炎(AIDP)的经典动物模型,本课题应用DHEA治疗发病初期EAN,通过观察EAN的临床和病理及免疫指标的改变,评价DHEA对发病初期EAN的治疗作用并探讨其免疫机制,为探索治疗自身免疫性疾病的新模式提供了依据。
创新性
1.本课题应用牛周围神经髓磷脂与完全弗氏佐剂混合制成抗原乳剂,注入易感鼠种Lewis大鼠双后肢足垫皮下,诱导实验性自身免疫性神经炎(EAN)动物模型。该模型是GBS最常见的临床亚型AIDP的经典动物模型,目前国内未见有应用上述方法及鼠种诱导EAN的报道。
2.本课题应用脱氢表雄酮(DHEA)治疗发病初期EAN,观察比较临床评分、坐骨神经炎性细胞浸润,评价DHEA的治疗效果。通过观察DHEA对引流淋巴结和脾脏单个核细胞体外增殖、培养上清IFN-γ、TNF-α、IL-10水平及坐骨神经中IFN-γ、TNF-α表达水平的影响,探讨其免疫治疗机制。目前国内外尚未见DHEA治疗EAN及其机制的研究报道。
Objective
Guillain-Barre syndrome(GBS) is the main cause of crippling disease in humans and characterized as acute demyelinating inflammation in structures of the peripheral nervous system(PNS).Experimental autoimmune neuritis(EAN) is a T cell-mediated,acute,autoimmune demyelination disease of the PNS that serves as a model for human GBS. The close clinical,histopathological,and electrophysiological similarities between EAN and GBS make EAN a classic animal model for studying pathogenic mechanisms and novel treating strategies for GBS. For the treatment of GBS,both plasma exchange and intravenous immunoglobulin have shown the efficacy in shorten the progressive course.But there is a mortality of approximately 5%,and approximately 20%of patients are left with significant and persistent disability.Many of the conventional immunosuppressive treatments,such as corticosteroids, have no effect on GBS.It is needed to uncover new therapeutic modalities for GBS.
Dehydroepiandrosterone(DHEA),a C19 adrenal steroid,is a precursor of both androgens and oestrogens.DHEA also serves as a precursor for many less well understood immune-regulating steroids. Studies showed that DHEA and its metabolites have anti-inflammatory, anti-proliferative,and certain immune-regulating properties both in vitro and in vivo.DHEA have shown beneficial effects on immunologic system in experimental animals and humans.EAN is a Thl-mediated autoimmune demyelination disease of the PNS.The infiltration of T cells and macrophages in the PNS and the multifocal,segmental demyelination are the main pathological changes of EAN,in which inflammatory cytokines, such as IFN-γ、TNF-α、IL-12、NO,play important roles.Thus far,there is no report about the role of DHEA in EAN.Based on the previous findings of DHEA on immunologic system,DHEA was used in treatment of EAN in the initial phase in this study.The therapeutical effect of DHEA on EAN and the immunologic mechanism were evaluated by examining the changes in clinical,histopathological and immunological parameters. Methods
1.Antigens:Bovine peripheral nerve myelin(BPM) was prepared from intra- and extradural spinal nerve roots by ultracentrifugation.
2.Animals:Thirty-six female Lewis rats,6-8 weeks old,weighing 150 - 180g were used in the present study.All of the rats were maintained in specific- pathogen free condition.
3.Induction and clinical evaluation of EAN:Ketamine was used for the animal anesthesia(10mg/Kg).EAN was induced by subcutaneous(s.c.) injection into both hind footpads with 100μl of inoculum containing 5 mg of BPM and 2 mg of Mycobacterium tuberculosis and 50μl Freund's incomplete adjuvant.Clinical scores were assessed immediately before immunization and thereafter every day until day 28 post immunization (p.i.).Severity of paresis was graded as follows:0= no illness;1= flaccid tail;2= dragging both hind limbs;3= paralysis of both hind limbs;4= paralysis of four limbs or death;intermediate scores of 0.5 increment were given to rats with intermediate signs.
4.DHEA treatment:Thirty-six Lewis rats were randomly divided into DHEA 0.5mg treatment group,2mg treatment group and control group(n = 12).Treatment groups were subcutaneously injected every day with DHEA and control group were subcutaneously injected with the same volum of DHEA dissolvent DMSO from day 5 p.i.to day 28 p.i.
5.Histopathological assessment:Seven rats from each group were killed on day 16 p.i.,at a time when the clinical signs of EAN peaked. Sciatic nerve segments were excised close to the lumbar spinal cord, fixed in 10%phosphate-buffered formalin,and embedded in paraffin. Multiple longitudinal sections(5- 6μm slices) of sciatic nerves were stained with hematoxylin and eosin for evaluation of the extent of mononuclear cell(MNC) infiltration by light microscopy.Tissue areas were measured by image analysis and the numbers of inflammatory cells were counted at×40 magnification.The average results were expressed as cells per mm~2 tissue section.
6.Immunohistochemistry:Paraffin tissue sections(5μm) were deparaffinized,hydrated and treated for blocking endogenous peroxidase activity,repairing antigen.The sections were incubated with polyclonal goat anti-rat IFN-γ、TNF-αantibody for 1h and then stained according to the avidin-biotin technique.The numbers of positive cells were counted at×40 magnification in the entire section area.The average results were expressed as cells per mm~2 tissue section.
7.Lymphocyte proliferation assay:The draining lymph nodes and spleens were removed under aseptic conditions.Single cell suspensions of MNC from individual rats were prepared separately.The cells were suspended in RPMI1640 and cultured in triplicates in round-bottomed 96-well microtitre plates in the presence of 10μl of BPM or phytohemagglutinin or the same volume of PBS.After 48h of incubation, proliferation was measured by labeling cells with[~3H]-methylthymidine (1μCi/well) and cultured for an additional 18h.Cells were harvested onto glass fibre filters.[~3H]-methylthymidine incorporation was measured in liquidβ- scintillation counter.Data are given as mean counts per minute (cpm).
8.Measurement of cytokine:MNC were cultured at a cell density of 2×10~6 cells/ml in medium containing BPM(20μg/ml).Supernatants were collected after 48h of incubation at 37℃.Quantitative ELISA assays for cytokines of IFN-γ,TNF-αand IL-10 were performed using ELISA kits following the manufacturer's instructions.Determinations were performed in duplicate and results were expressed as pg/ml.
9.Statistical analysis:The SPSS 13.0 computer program was used for all calculations and statistical evaluations.Results were expressed as means±standard deviation(SD).Differences between groups were evaluated by one-factor analysis of variance(ANOVA) or rank-sum test. The level of significance was set to p＜0.05.
Results
1.DHEA treatment delayed disease onset of EAN:Both DHEA-treated groups exhibited more delayed disease onset than control rats(P＜0.05, P＜0.05).Compared to 0.5 mg DHEA treatment group,2 mg group also displayed significant delayed disease onset(P＜0.05).
2.DHEA treatment decreased disease peak clinical scores of EAN: The rats treated with DHEA at dose of 2 mg/day showed significantly lower mean peak clinical scores than the control rats(P＜0.05).0.5 mg DHEA treatment group showed lower mean peak clinical scores than the control group,but there was no statistically significant difference.
2.DHEA treatment inhibits inflammatory cell infiltration of PNS:The sciatic nerves of EAN rats were examined histologically at the height of the clinical course of EAN.The results revealed that administration of DHEA at doses of both 2 mg/day and 0.5 mg/day all significantly reduced the infiltrating cells in sciatic nerves sections compared to control EAN rats(P＜0.05,P＜0.05).2 mg DHEA treatment group showed less numbers of infiltrating cells in sciatic nerves than 0.5 mg treatment group,but there was no significant difference statistically.
3.Effects of DHEA on MNCs Proliferation:MNCs of draining lymph nodes and spleens from EAN rats were tested on day 16 p.i.for lymphocytes proliferation.When compared to the controls,the DHEA treated rats showed significantly suppressed BPM and PHA-induced MNC proliferation(P＜0.05,P＜0.05,P＜0.05,P＜0.05 ).In addition,there was also a lower level of proliferation in 2 mg DHEA treatment rats than control EAN rats when MNC were cultured in the absence of antigen(P＜0.05),but there was no statistically significant difference between 0.5 mg DHEA treatment group and the controls.
4.DHEA treatment suppresses the cytokine expression in sciatic nerves:Immunohistochemical studies showed that the numbers of IFN-γand TNF-α.expressing cells were strongly decreased in the sciatic nerves of DHEA treated rats at dose of 2 mg/day and 0.5 mg/day compared to control EAN rats(P＜0.05,P＜0.05,P＜0.05,P＜0.05 ).No statistically confirmed difference regarding the numbers of cells expressing IFN-γ,and TNF-αin the PNS was shown when comparing 2 mg-DHEA to 0.5 mg -DHEA treatment groups.
5.Levels of cytokine in supernatant:The levels of IFN-γ,TNF-αand IL-10 secretion from MNC supernatants were examined by ELISA.The results showed that levels of IFN-γ,and TNF-α.in the MNC supernatant were significantly decreased in EAN rats treated with DHEA at doses of 2 mg/day and 0.5 mg/day when examined on day 16 p.i.as compared with control EAN rats(P＜0.05,P＜0.05,P＜0.05,P＜0.05).There were also statistically significant differences in IFN-γ,and TNF-αsecretion in the MNC supernatants between 2 mg DHEA and 0.5 mg DHEA treated EAN rats(P＜0.05,P＜0.05).There were no differences in IL-10 production among the three groups of rats.
Conclusion
1.In this study,DHEA was administered to EAN rats at doses of 0.5 mg/rat/day and 2 mg/rat/day for the treatment of EAN in the initial phase. Rats treated with DHEA at the both doses displayed significant delay in onset,lower peak clinical score and decreased inflammatory cell infiltration in the PNS as compared to control rats.These data suggest that DHEA has therapeutic potential for alleviating EAN.
2.Rats treated with DHEA at the both doses displayed significant decreases in numbers of IFN-γ,and TNF-αexpressing cells in the PNS, BPM-stimulated MNCs proliferation and IFN-γ,TNF-α-secretion in the draining lymph node and spleen cells.Our finding suggests that DHEA could inhibit cellular immunological response by suppressing the proliferation of autoreactive T cells and overexpression of proinflamation cytokines.
3.In the present study,administration of DHEA to EAN rats decreased IFN-γ、TNF-αsecretion both systemically and within sites of inflammation,with no significant changes in IL-10 production.Our finding suggests that DHEA suppressed EAN mainly by downregulation of autoimmune Th1 response,decreasing the production of proinflammatory cytokines without changing levels of Th2 associated cytokines.
4.Our data show that rats treated with DHEA at dose of 2mg every day exhibited delayed disease onset,lower mean peak clinical scores and decreased levels of IFN-γ,and TNF-α.from cultured MNC supernatants when compared to 0.Smg DHEA-treated EAN rats.The results suggest that the immune suppressive effect of DHEA is dose-dependent.
Significance
Guillain-Barre syndrome(GBS) is the most main cause of crippling disease in humans and characterized as acute demyelinating inflammation in structures of the peripheral nervous system.For the treatment of GBS, both plasma exchange and intravenous immunoglobulin have shown the efficacy in shorten the progressive course and hastening recovery from GBS.But there is a mortality of approximately 5%,and approximately 20%of patients are left with significant and persistent disability.Further research is needed to uncover new therapeutic modalities for GBS to block the cascade inflammation and decreas the mortality and sequelae of nervous system.
Dehydroepiandrosterone(DHEA) is the most abundant circulating adrenal steroids in humans.It has been well-documented that DHEA and its metabolites have anti-inflammatory,anti-proliferative,and certain immune-regulating properties and have shown immunological effects both in vitro and in vivo in experimental animals and humans.Experimental autoimmune neuritis(EAN) serves as classic animal model for acute inflammatory demyelinative polyradiculoneuropathy(AIDP) - one of the most common subtype of human.In this study,the therapeutical effect of DHEA on the initial phase of EAN and the immunologic mechanism were evaluated by examining the changes in clinical,histopathological and immunological parameters.We aimed to provide evidence for searching a promising new and potentially powerful strategy for human autoimmune disorders.
creativity
1.In our study,the inoculum was prepared by emulsifying bovine peripheral nerve myelin(BPM) in Freund's complete adjuvant(CFA). EAN was induced by subcutaneous injection the inoculum into both hind footpads of susceptible animals - Lewis rat.EAN serves as a classical model for human AIDP,which is the most common subtype of GBS.So far,there has been no report on inducing EAN with BPM and CFA in Lewis rat in China.
2.In this study,DHEA was used for the treatment of EAN in the initial phase.The therapeutic effect of DHEA was evaluated by assessing clinical scores and examining inflammatory cell infiltration in the PNS. The immunologic mechanism were evaluated by examining the changes in BPM-stimulated MNCs proliferation from draining lymph nodes and spleens,IFN-γ,TNF-β,IL-10 -secretion from MNC supernatants and IFN-γ,TNF-αexpressing cells in the PNS.Up to now,there has been no report about DHEA administration for the treatment of EAN at home and abroad.
引文
1 Kuwabara S. Guillain-barre syndrome. Curr Neurol Neurosci Rep. 2007;7:57-62..
2 Hughes RA, Cornblath DR.Guillain-Barre syndrome. Lancet. 2005; 366: 1653-66.
3 Svec F, Porter JR.The actions of exogenous dehydroepiandrosterone in experimental animals and humans.Proc Soc Exp Biol Med. 1998; 218:174-91.
4 von Mühlen D, Laughlin GA, Kritz-Silverstein D, et al. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods. Contemp Clin Trials. 2007;28:153-68.
5 Oberbeck R, Dahlweid M, Koch R,et al. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29 :380-4.
6 Padgett DA, Loria RM, Sheridan JF. Steroid hormone regulation of antiviral immunity. Ann N Y Acad Sci. 2000;917:935-43.
7 Loria RM. Immune up-regulation and tumor apoptosis by androstene steroids. Steroids. 2002; 7: 53-66.
8 Loria RM, Conrad DH, Huff T, et al. Androstenetriol and androstenediol. Protection against lethal radiation and restoration of immunity after radiation injury.Ann N Y Acad Sci. 2000;917:860-7.
9 Yu CK, Yang BC, Lei HY, et al.Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response.Clin Exp Allergy. 1999;29:414-22.
10 Du C, Guan Q, Khalil MW, et al..Stimulation of Th2 response by high doses of dehydroepiandrosterone in KLH-primed splenocytes. Exp Biol Med (Maywood). 2001;226:1051-60.
11 Powell JM, Sonnenfeld G.The effects of dehydroepiandrosterone (DHEA) on in vitro spleen cell proliferation and cytokine production. J Interferon Cytokine Res. 2006;26:34-9.
12 Du C, Khalil MW, Sriram S. Administration of dehydro-epiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice.J Immunol. 2001; 167: 7094-101.
13 Yang BC, Liu CW, Chen YC, et al. Exogenous dehydro-epiandrosterone modified the expression of T helper-related cytokines in NZB/NZW F1 mice.Immunol Invest. 1998;27:291-302.
14 van Vollenhoven RF, Morabito LM, Engleman EG, et al.Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J Rheumatol. 1998;25:285-9.
15 van Vollenhoven RF. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3:23-31.
16 Straub RH, Scholmerich J, Zietz B.Replacement therapy with DHEA plus corticosteroids in patients with chronic inflammatory diseases-substitutes of adrenal and sex hormones.Z Rheumatol. 2000;59 Suppl 2:11/108-18.
17 Maurer M,Gold R.Animal models of immune-mediated neuropathies. Curr Opin Neurol. 2002;15:617-22.
1 Maurer M, Gold R. Animal models of immune-mediated neuropathies. Curr Opin Neurol. 2002 ;15:617-22.
2 Gold R, Hartung HP, Toyka KV.Animal models for autoimmune demyelinating disorders of the nervous system.Mol Med Today. 2000; 6:88-91.
3 Norton WT, Poduslo SE.Myelination in rat brain: method of myelin isolation. J Neurochem. 1973 ;21:749-57. FEBS Lett.
4 Kitamura K, Suzuki M, Suzuki A,et al.The complete amino acid sequence of the P2 protein in bovine peripheral nerve myelin. FEBS Lett. 1980 Jun 16; 115:27-30.
5 Uyemura K, Suzuki M, Kitamura K, et al. Neuritogenic determinant of bovine P2 protein in peripheral nerve myelin. J Neurochem. 1982;39:895-8.
6 Kuwabara S. Guillain-barre syndrome. Curr Neurol Neurosci Rep. 2007; 7: 57-62.
7 Hughes RA, Cornblath DR.Guillain-Barre syndrome. Lancet. 2005; 366: 1653-66.
8 Waksman BH, Adams RD. Allergic neuritis: an experimental disease of rabbits induced by the injection of peripheral nervous tissue and adjuvants.J Exp Med. 1955; 102:213-36.
9 Milner P, Lovelidge CA, Taylor WA,et al.PO myelin protein produces experimental allergic neuritis in Lewis rats. J Neurol Sci. 1987;79:275-85.
10 Ben-Nun A, Wekerle H, Cohen IR.The rapid isolation of clonable antigen-specific T lymphocyte lines capable of mediating autoimmune encephalomyelitis.Eur J Immunol. 1981 ;11:195-9.
11 Olee T, Powell HC, Brostoff SW. New minimum length requirement for a T cell epitope for experimental allergic neuritis. J Neuroimmunol. 1990; 27: 187-90
12 Taylor WA, Hughes RA.Experimental allergic neuritis induced in SJL mice by bovine P2.J Neuroimmunol. 1985; 8:153-7.
13 Calida DM, Kremlev SG, Fujioka T,et al.Experimental allergic neuritis in the SJL/J mouse: induction of severe and reproducible disease with bovine peripheral nerve myelin and pertussis toxin with or without interleukin-12. J Neuroimmunol. 2000;107:l-7.
14 Zou LP, Ljunggren HG, Levi M, et al.PO protein peptide 180-199 together with pertussis toxin induces experimental autoimmune neuritis in resistant C57BL/6 mice.J Neurosci Res. 2000;62:717-21.
15 Zhu J, Zou L, Zhu S,et al.Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) blockade enhances incidence and severity of experimental autoimmune neuritis in resistant mice.J Neuroimmunol. 2001;115:111-7.
1 Miller RA. Aging and immune function.Int Rev Cytol. 1991; 124: 187-215.
2 Svec F, Porter JR.The actions of exogenous dehydroepiandrosterone in experimental animals and humans.Proc Soc Exp Biol Med. 1998 ;218:174-91.
3 Oberbeck R, Dahlweid M, Koch R,et al. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29:380-4.
4 Small M, Gray CE, Beastall GH, et al. Adrenal androgens in insulin-dependent diabetes mellitus, Diabetes Res. 1989;11:93-5.
5 Henderson E, Schwartz A, Pashko L,et al. Dehydroepiandrosterone and 16 alpha-bromo-epiandrosterone: inhibitors of Epstein-Barr virus- induced transformation of human lymphocytes. Carcinogenesis. 1981; 2:683-6.
6 Khaw KT. Dehydroepiandrosterone, dehydroepiandrosterone sulphate and cardiovascular disease.J Endocrinol. 1996 ;150 Suppl: S149-53.
7 Du C, Khalil MW, Sriram S.Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice.J Immunol. 2001; 167 :7094-101.
8 Rontzsch A, Thoss K, Petrow PK,et al. Amelioration of murine antigen- induced arthritis by dehydroepiandrosterone (DHEA). Inflamm Res. 2004;53:189-98.
9 van Vollenhoven RF. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002;3: 23-31.
10 Dillon JS. Dehydroepiandrosterone, Dehydroepiandrosterone Sulfate and Related Steroids: Their Role in Inflammatory, Allergic and Immunological Disorders. Curr Drug Targets Inflamm Allergy. 2005;4:377-85.
11 Stephanie JW, Thomas EG, Russell AP. The biological action of Dehydroepiandrosterone involves multiple receptors. Drug Metabolism. 2006;38: 89-116.
12 von Mühlen D, Laughlin GA, Kritz-Silverstein D, et al. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods. Contemp Clin Trials. 2007;28:153-68.
13 Dessein PH, Joffe BI, Stanwix AE, et al.Hyposecretion of the adrenal androgen dehydroepiandrosterone sulfate and its relation to clinical variables in inflammatory arthritis.Arthritis Res. 2001 ;3:183-8.
14 Khalkhali-Ellis Z, Moore TL, Hendrix MJ.Reduced levels of testosterone and dehydroepiandrosterone sulphate in the serum and synovial fluid of juvenile rheumatoid arthritis patients correlates with disease severity.Clin Exp Rheumatol. 1998;16:753-6.
15 Straub RH, Zeuner M, Antoniou E, et al. Dehydroepiandrosterone sulfate is positively correlated with soluble interleukin 2 receptor and soluble intercellular adhesion molecule in systemic lupus erythematosus.J Rheumatol. 1996;23:856-61.
16 de la Torre B, von Krogh G, Svensson M, et al. Blood cortisol and dehydroepiandrosterone sulphate (DHEAS) levels and CD4 T cell counts in HIV infection. Clin Exp Rheumatol. 1997;15:87-90.
17 Henderson E, Yang JY, Schwartz A Dehydroepiandrosterone (DHEA) and synthetic DHEA analogs are modest inhibitors of HIV-1 IIIB replication. AIDS Res Hum Retroviruses. 1992; 8:625-31.
18 van Vollenhoven RF, Morabito LM, Engleman EG, et al. Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J Rheumatol. 1998;25:285-9.
19 van Vollenhoven RF. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3: 23-31.
20 Svec F, Porter JR.The actions of exogenous dehydroepiandrosterone in experimental animals and humans.Proc Soc Exp Biol Med. 1998; 218: 174- 91.
21 Oberbeck R, Dahlweid M, Koch R,et al. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29:380-4.
22 Padgett DA, Loria RM, Sheridan JF. Steroid hormone regulation of antiviral immunity. Ann N Y Acad Sci. 2000;917:935-43.
23 Loria RM. Immune up-regulation and tumor apoptosis by androstene steroids. Steroids. 2002 ; 7: 53-66.
24 Loria RM, Conrad DH, Huff T,et al. Androstenetriol and androstenediol. Protection against lethal radiation and restoration of immunity after radiation injury.Ann N Y Acad Sci. 2000;917:860-7.
25 Du C, Khalil MW, Sriram S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice.J Immunol. 2001;167: 7094-101.
26 Rontzsch A, Thoss K, Petrow PK, et al. Amelioration of murine antigen- induced arthritis by dehydroepiandrosterone (DHEA). Inflamm Res. 2004; 53: 189-98.
27 Offner H, Zamora A, Drought H, et al.A synthetic androstene derivative and a natural androstene metabolite inhibit relapsing-remitting EAE. J Neuroimmunol. 2002;130:128-39.
28 Offner H, Zamora A, Subramanian S, et al. A synthetic androstene analogue inhibits collagen-induced arthritis in the mouse. Clinical Immunology. 2004;l 10:181-90.
29 Yang BC, Liu CW, Chen YC, et al.Exogenous dehydroepiandrosterone modified the expression of T helper-related cytokines in NZB/NZW Fl mice.Immunol Invest. 1998;27:291-302.
30 Duan, R.S., Link, H., Xiao, B.G. Dehydroepiandrosterone therapy ameliorates experimental autoimmune myasthenia gravis in Lewis rats. J Clin Immunol. 2003;23:100-6.
31 Yoneda M, Wada K, Katayama K, et al. A novel therapy for acute hepatitis utilizing dehydroepiandrosterone in the murine model of hepatitis. Biochem Pharmacol. 2004;68:2283-9.
32 Auci D, Nicoletti F, Mangano K, et al.Anti-inflammatory and Immune Regulatory Properties of 5-Androsten-3_, 17_-Diol (HE2100), and Synthetic Analogue HE3204 Implications for Treatment of Autoimmune Diseases. Ann. N.Y. Acad. Sci. 2005;1051: 730-742.
33 Kipper-Galperin M, Galilly R, Danenberg HD, et al. Dehydroepiandrosterone selectively inhibits production of tumor necrosis factor-a and interlukin-6 in astrocytes. Int. J. Dev. Neurosci. 1999;17:765-75.
1 Hughes RA, Cornblath DR.Guillain-Barre syndrome. Lancet. 2005; 366:1653-66.
2 Kuwabara S. Guillain-barre syndrome. Curr Neurol Neurosci Rep. 2007; 7: 57-62.
3 Svec F, Porter JR.The actions of exogenous dehydroepiandrosterone in experimental animals and humans.Proc Soc Exp Biol Med. 1998; 218: 174- 91.
4 von Mühlen D, Laughlin GA, Kritz-Silverstein D, et al. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods[J]. Contemp Clin Trials, 2007,28:153-168.
5 Straub RH, Vogl D, Gross V,et al. Association of humoral markers of inflammation and dehydroepiandrosterone sulfate or cortisol serum levels in patients with chronic inflammatory bowel disease.Am J Gastroenterol. 1998; 93:2197-202.
6 Straub RH, Paimela L, Peltomaa R, et L.Inadequately low serum levels of steroid hormones in relation to interleukin-6 and tumor necrosis factor in untreated patients with early rheumatoid arthritis and reactive arthritis.Arthritis Rheum. 2002;46:654-62.
7 Straub RH, Lehle K, Herfarth H, et al.Dehydroepiandrosterone in relation to other adrenal hormones during an acute inflammatory stressful disease state compared with chronic inflammatory disease: role of interleukin-6 and tumour necrosis factor.Eur J Endocrinol. 2002;146:365-74.
8 Suzuki T, Suzuki N, Engleman EG, et al. Low serum levels of dehydroepiandrosterone may cause deficient IL-2 production by lymphocytes in patients with systemic lupus erythematosus (SLE).Clin Exp Immunol. 1995; 99:251-5.
9 Straub RH, Zeuner M, Antoniou E, et al. Dehydroepiandrosterone sulfate is positively correlated with soluble interleukin 2 receptor and soluble intercellular adhesion molecule in systemic lupus erythematosus.J Rheumatol. 1996;23:856-61.
10 Simard J, Gingras S. Crucial role of cytokines in sex steroid formation in normal and tumoral tissues.Mol Cell Endocrinol. 2001;171:25-40.
11 Turgeon C, Gingras S, Carriere MC, et al. Regulation of sex steroid formation by nterleukin-4 and interleukin-6 in breast cancer cells.J Steroid Biochem Mol Biol. 1998;65:151-62.
12 Tabata N, Tagami H, Terui T. Dehydroepiandrosterone may be one of the regulators of cytokine production in atopic dermatitis.Arch Dermatol Res. 1997;289:410-4.
13 Hernandez-Pando R, De La Luz Streber M, Orozco H, et al. The effects of androstenediol and dehydroepiandrosterone on the course and cytokine profile of tuberculosis in BALB/c mice.Immunology. 1998;95:234-41.
14 Suzuki T, Suzuki N, Daynes RA, et al. Dehydroepiandrosterone enhances IL2 production and cytotoxic effector function of human T cells.Clin Immunol Immunopathol. 1991 ;61:202-11.
15 Canning MO, Grotenhuis K, de Wit HJ, et al.Opposing effects of dehydroepiandrosterone and dexamethasone on the generation of monocyte-derived dendritic cells. Eur J Endocrinol. 2000; 143: 687-95.
16 Zhang Z, Araghi-Niknam M, Liang B, et al.Prevention of immune dysfunction and vitamin E loss by dehydroepiandrosterone and melatonin supplementation during murine retrovirus infection. Immunology. 1999;96:291-7
17 Yu CK, Yang BC, Lei HY,et al. Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response. Clin Exp Allergy. 1999;29:414-22
18 Du C, Guan Q, Khalil MW, et al.Stimulation of Th2 response by high doses of dehydroepiandrosterone in KLH-primed splenocytes.Exp Biol Med (Maywood). 2001 Dec;226:1051-60.
19 Powell JM, Sonnenfeld G.The effects of dehydroepiandrosterone (DHEA) on in vitro spleen cell proliferation and cytokine production. J Interferon Cytokine Res. 2006;26:34-9.
20 Du C, Khalil MW, Sriram S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice.J Immunol. 2001 ;167:7094-101.
21 Yang BC, Liu CW, Chen YC, et al . Exogenous dehydroepiandrosterone modified the expression of T helper-related cytokines in NZB/NZW F1 mice.
Immunol Invest. 1998;27(4-5):291-302.
22 Di Santo E, Sironi M, Mennini T, et al. A glucocorticoid receptor-independent mechanism for neurosteroid inhibition of tumor necrosis factor production.Eur J Pharmacol. 1996;299:179-86.
23 Rontzsch A, Thoss K, Petrow PK, Henzgen S, Brauer R. Amelioration of murine antigen-induced arthritis by dehydroepiandrosterone (DHEA). Inflamm Res. 2004;53:189-98.
24 Offner H, Zamora A, Drought H, et al.A synthetic androstene derivative and a natural androstene metabolite inhibit relapsing-remitting EAE.J Neuroimmunol. 2002; 130:128-39.
25 Offner H, Zamora A, Subramanian S, et al. A synthetic androstene analogue inhibits collagen-induced arthritis in the mouse. Clin Immunol. 2004; 110: 181-90.
26 van Vollenhoven RF, Morabito LM, Engleman EG, et al.Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J Rheumatol. 1998;25:285-9.
27 van Vollenhoven RF. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3: 23-31.
28 Giltay EJ, van Schaardenburg D, Gooren LJ, et al. Dehydro-
epiandrosterone sulfate in patients with rheumatoid arthritis.Ann N Y Acad Sci. 1999;876:152-4.
29 Labrie F, Luu-The V, Labrie C, et al.DHEA and its transformation into androgens and estrogens in peripheral target tissues: intracrinology.Front Neuroendocrinol. 2001;22:185-212.
30 Sugiyama H, Yamada J, Takama H, et al.Photoaffinity labeling of peroxisome proliferator binding proteins in rat hepatocytes; dehydroepiandrosterone sulfate- and bezafibrate-binding proteins. Biochim Biophys Acta. 1997; 1339: 321-30.
31 Tsuji K, Furutama D, Tagami M, et al.Specific binding and effects of dehydroepiandrosterone sulfate (DHEA-S) on skeletal muscle cells: possible implication for DHEA-S replacement therapy in patients with myotonic dystrophy.Life Sci. 1999;65:17-26.
32 Bruder JM, Sobek L, Oettel M. Dehydroepiandrosterone stimulates the estrogen response element. J Steroid Biochem Mol Biol. 1997;62:461-6.
33 Tan J, Sharief Y, Hamil KG, et al.Dehydroepiandrosterone activates mutant androgen receptors expressed in the androgen-dependent human prostate cancer xenograft CWR22 and LNCaP cells.Mol Endocrinol. 1997;11(4): 450-9.
34 Peters JM, Zhou YC, Ram PA,et al. Peroxisome proliferator-activated receptor alpha required for gene induction by dehydroepiandrosterone- 3beta-sulfate. Mol Pharmacol. 1996;50:67-74.
35 Poynter ME, Daynes RA. Peroxisome proliferator-activated receptor alpha activation modulates cellular redox status, represses nuclear factor-kappaB signaling, and reduces inflammatory cytokine production in aging. J Biol Chem. 1998;273:32833-41.
36 Aragno M, Parola S, Tamagno E, et al. Oxidative derangement in rat synaptosomes induced by hyperglycaemia: restorative effect of dehydroepiandrosterone treatment.Biochem Pharmacol. 2000 Aug l;60:389-95.
37 Zhu Y, Ljunggren HG, Mix E, et al. Suppression of autoimmune neuritis in IFN-gamma receptor-deficient mice. Experimental Neurology. 2001; 169: 472-478.
38 Weishaupt A, Bruck W, Hartung T, et al. Schwann cell apoptosis in experimental autoimmune neuritis of the Lewis rat and the functional role of tumor necrosis factor-alpha. Neurosci Lett, 2001, 306: 77-80.
39 Dillon JS. Dehydroepiandrosterone, dehydroepiandrosterone sulfate and related steroids: their role in inflammatory, allergic and immunological disorders. Curr Drug Targets Inflamm Allergy, 2005,4:377-385.
1 Maurer M., Gold R. Animal models of immune-mediated neuropathies. Curr Opin Neurol. 2002; 15, 617-622.
2 Hughes R.A., Cornblath, D.R(?) Guillain-Barre syndrome. Lancet. 2005; 366, 1653-1666.
3 Kuwabara S., Guillain-barre syndrome. Curr Neurol Neurosci Rep. 2007;7, 57-62.
4 von Muhlen D., Laughlin G.A., Kritz-Silverstein D., et al. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods. Contemp Clin Trials. 2007;28, 153-168.
5 Svec F., Porter J.R. The actions of exogenous dehydroepiandrosterone in experimental animals and humans. Proc Soc Exp Biol Med. 1998;218, 174-191.
6 Oberbeck R., Dahlweid M., Koch R.,.et al. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29, 380-384.
7 Yu C.K., Yang B.C., Lei H.Y., et al. Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response. Clin Exp Allergy. 1999;29, 414-22.
8 Du C., Guan Q., Khalil M.W., et al. Stimulation of Th2 response by high doses of dehydroepiandrosterone in KLH-primed splenocytes. Exp Biol Med (Maywood). 2001;226, 1051-1060.
9 Powell J.M., Sonnenfeld G. The effects of dehydroepiandrosterone (DHEA) on in vitro spleen cell proliferation and cytokine production. J Interferon Cytokine Res. 2006;26, 34-39.
10 Du C., Khalil M.W., Sriram S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice. J Immunol. 2001;167, 7094-7101.
11 Yang B.C., Liu C.W., Chen Y.C., et al. Exogenous dehydroepiandrosterone modified the expression of T helper-related cytokines in NZB/NZW F1 mice. Immunol Invest. 1998;27, 291-302.
12 Straub R.H., Scholmerich J., Zietz B. Replacement therapy with DHEA plus corticosteroids in patients with chronic inflammatory diseases--substitutes of adrenal and sex hormones. Z. Rheumatol. 2000; 59 Suppl 2:II, 108-118.
13 van Vollenhoven R.F. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3, 23-31.
14 van Vollenhoven R.F., Morabito L.M., Engleman E.G., et al. Treatment of systemic lupus erythematosus with dehydro- epiandrosterone: 50 patients treated up to 12 months. J. Rheumatol. 1998; 25, 285-289.
15 Norton W.T., Poduslo S.E. Myelination in rat brain: method of myelin isolation. J. Neurochem. 1973; 21,749-757.
16 Auci D., Nicoletti F., Mangano K.,et al. Anti-inflammatory and Immune Regulatory Properties of 5-Androsten-3_β 17_β-Diol (HE2100), and Synthetic Analogue HE3204 Implications for Treatment of Autoimmune Diseases. Ann. N.Y. Acad. Sci. 2005;1051, 730-742.
17 Duan R.S., Link H., Xiao B.G. Dehydroepiandrosterone therapy ameliorates experimental autoimmune myasthenia gravis in Lewis rats. J Clin Immunol. 2003;23, 100-106.
18 Offner H., Zamora A., Drought H., et al. A synthetic androstene derivative and a natural androstene metabolite inhibit relapsing-remitting EAE. J. Neuroimmunol. 2002;30, 128-139.
19 Offner H., Zamora A., Subramanian S., et al. synthetic androstene analogue inhibits collagen-induced arthritis in the mouse. Clin Immunol. 2004; 110, 181-190.
20 Rontzsch A., Thoss K., Petrow P.K., et al. Amelioration of murine antigen-induced arthritis by dehydroepiandrosterone (DHEA). Inflamm Res. 2004;53, 189-198.
21 Yoneda M., Wada K., Katayama K., et al. A novel therapy for acute hepatitis utilizing dehydroepiandrosterone in the murine model of hepatitis. Biochem Pharmacol. 2004;68,2283-2289.
22 Kipper-Galperin M., Galilly R., Danenberg H.D., et al. Dehydroepiandrosterone selectively inhibits production of tumor necrosis factor-a and interlukin-6 in astrocytes. Int. J. Dev. Neurosci. 1999;17, 765-775.
23 Zhu, J., Mix, E., Link, H. Cytokine production and the pathogenesis of experimental autoimmune neuritis and Guillain-Barre syndrome. J. Neuroimmunol. 1998; 84, 40-52.
24 Zhu Y., Ljunggren H.G., Mix E., et al. Suppression of autoimmune neuritis in IFN-gamma receptor-deficient mice. Exp Neurol. 2001;169, 472-478.
25 Redford E.J., Hall S.M., Smith K.J. Vascular changes and demyelination induced by the intraneural injection of tumour necrosis factor. Brain. 1995;118, 869-878.
26 Weishaupt A., Bruck W., Hartung T., et al. Schwann cell apoptosis in experimental autoimmune neuritis of the Lewis rat and the functional role of tumor necrosis factor-alpha. Neurosci Lett. 2001; 306, 77-80.
27 Elkarim, R.A., Dahle, C., Mustafa, M.,et al. Recovery from Guillain-Barre syndrome is associated with increased levels of neutralizing autoantibodies to interferon-gamma. Clin Immunol Immunopathol. 1998;88, 241-248.
28 Hohnoki, K., Inoue, A., Koh, C.S. Elevated serum levels of IFN-gamma, IL-4 and TNF-alpha/unelevated serum levels of IL-10 in patients with demyelinating diseases during the acute stage. J. Neuroimmunol. 1998; 87, 27-32.
29 Misawa, S., Kuwabara, S., Mori, M., et al. Serum levels of tumor necrosis factor-alpha in chronic inflammatory demyelinating polyneuropathy. Neurology. 2001; 56, 666-669.
30 Radhakrishnan, V.V., Sumi, M.G., Reuben, S., et al. Serum tumour necrosis factor-alpha and solubletumour necrosis factor receptors levels in patients withGuillain-Barre syndrome. Acta Neurol Scand . 2004;109, 71-74.
31 Sharief ,M.K., Ingram, D.A., Swash, M. Circulating tumor necrosis factor-alpha correlates with electrodiagnostic abnormalities in Guillain-Barre syndrome. Ann Neurol. 1997;42, 68-73.
32 Dillon,J.S. Dehydroepiandrosterone, Dehydroepiandrosterone Sulfate and Related Steroids: Their Role in Inflammatory, Allergic and Immunological Disorders. Curr Drug Targets Inflamm Allergy. 2005; 4,377-385.
1 Svec, F., Porter, J.R. The actions of exogenous dehydroepiandrosterone in experimental animals and humans. Proc Soc Exp Biol Med. 1998; 218, 174-191.
2. von Mühlen, D., Laughlin, G.A., Kritz-Silverstein, D., Barrett-Connor, E. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods. Contemp Clin Trials. 2007; 28, 153-168.
3 Oberbeck, R., Dahlweid, M., Koch, R., van Griensven, M., Emmendorfer, A., Tscherne, H., Pape, H.C. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29, 380-384.
4 Yu, C.K., Yang, B.C., Lei, H.Y., Chen, Y.C., Liu, Y.H., Chen, C.C., Liu, C.W. Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response. Clin Exp Allergy. 1999; 29, 414-22.
5 Du, C., Guan, Q., Khalil M.W., Sriram, S. Stimulation of Th2 response by high doses of dehydroepiandrosterone in KLH-primed splenocytes. Exp Biol Med (Maywood). 2001; 226, 1051-1060.
6 Straub, R.H., Scholmerich, J., Zietz, B. Replacement therapy with DHEA plus corticosteroids in patients with chronic inflammatory diseases—substitutes of adrenal and sex hormones. Z. Rheumatol. 2000;59 Suppl 2:11, 108-118.
7 van Vollenhoven, R.F. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3, 23-31.
8 van Vollenhoven, R.F., Morabito, L.M., Engleman, E.G., McGuire, J.L.Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J. Rheumatol. 1998;25, 285-289.
9 Maurer M, Gold R.Animal models of immune-mediated neuropathies. Curr Opin Neurol. 2002 ;15:617-22.
10 Gold R, Hartung HP, Toyka KV. Animal models for autoimmune demyelinating disorders of the nervous system. Mol Med Today. 2000; 6: 88-91.
11 Norton, W.T., Poduslo, S.E. Myelination in rat brain: method of myelin isolation. J. Neurochem. 1973 ;21,749-757.
12 Du C, Khalil MW, Sriram S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice. J Immunol. 2001,167:7094-7101.
13 Duan RS, Link H, Xiao BG. Dehydroepiandrosterone therapy ameliorates experimental autoimmune myasthenia gravis in Lewis rats. J Clin Immunol. 2003,23(2):100-106.
14 Shin HC, McFarlane EF, Pollard JD, Watson EG. Induction of experimental allergic neuritis with synthetic peptides from myelin P2 protein. Neurosci Lett 1989;102:309-12
15 Linington C, Izumo S, Suzuki M, Uyemura K, Meyermann R,Wekerle H. A permanent rat T cell line that mediates experimental allergic neuritis in the Lewis rat in vivo. J Immunol 1984; 133:1946-50
16 Linington C, Lassmann H, Ozawa K, Kosin S, Mongan L. Cell adhesion molecules of the immunoglobulin supergene family as tissuespecific autoantigens: Induction of experimental allergic neuritis (EAN) by P0 protein-specific T cell lines. Eur J Immunol 1992;22:1813-17
17 Hahn AF. Experimental allergic neuritis (EAN) as a model for the immune-mediated demyelinating neuropathies. Rev Neurol 1996;152:328—32
18 Du, C., Khalil, M.W., Sriram, S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice. J Immunol. 2001;167, 7094-7101.
19 Offner, H., Zamora, A., Drought, H., Matejuk, A., Auci, D.L., Morgan, E.E., Vandenbark, A.A., Reading, C.L. A synthetic androstene derivative and a natural androstene metabolite inhibit relapsing-remitting EAE. J. Neuroimmunol. 2002;30, 128-139.
20 Zhu, J., Mix, E., Link, H. Cytokine production and the pathogenesis of experimental autoimmune neuritis and Guillain-Barre syndrome. J. Neuroimmunol. 1998;84,40-52.
21 Zhu, Y., Ljunggren, H.G., Mix, E., Li, H.L., van der Meide, P., Elhassan, A.M., Winblad, B., Zhu, J. Suppression of autoimmune neuritis in IFN-gamma receptor-deficient mice. Exp Neurol. 2001;169, 472-478.
22 Redford, E.J., Hall, S.M., Smith, K.J. Vascular changes and demyelination induced by the intraneural injection of tumour necrosis factor. Brain. 1995; 118,869-878.
23 Weishaupt ,A., Bruck, W., Hartung, T., Toyka, K.V., Gold, R. Schwann cell apoptosis in experimental autoimmune neuritis of the Lewis rat and the functional role of tumor necrosis factor-alpha. Neurosci Lett. 2001;306, 77-80.
24 Dillon, J.S. Dehydroepiandrosterone, Dehydroepiandrosterone Sulfate and Related Steroids: Their Role in Inflammatory, Allergic and Immunological Disorders. Curr Drug Targets Inflamm Allergy. 2005; 4, 377-385.
2 Hughes RA, Cornblath DR.Guillain-Barre syndrome. Lancet. 2005; 366: 1653-66.
3 Svec F, Porter JR.The actions of exogenous dehydroepiandrosterone in experimental animals and humans.Proc Soc Exp Biol Med. 1998; 218:174-91.
4 von Mühlen D, Laughlin GA, Kritz-Silverstein D, et al. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods. Contemp Clin Trials. 2007;28:153-68.
5 Oberbeck R, Dahlweid M, Koch R,et al. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29 :380-4.
6 Padgett DA, Loria RM, Sheridan JF. Steroid hormone regulation of antiviral immunity. Ann N Y Acad Sci. 2000;917:935-43.
7 Loria RM. Immune up-regulation and tumor apoptosis by androstene steroids. Steroids. 2002; 7: 53-66.
8 Loria RM, Conrad DH, Huff T, et al. Androstenetriol and androstenediol. Protection against lethal radiation and restoration of immunity after radiation injury.Ann N Y Acad Sci. 2000;917:860-7.
9 Yu CK, Yang BC, Lei HY, et al.Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response.Clin Exp Allergy. 1999;29:414-22.
10 Du C, Guan Q, Khalil MW, et al..Stimulation of Th2 response by high doses of dehydroepiandrosterone in KLH-primed splenocytes. Exp Biol Med (Maywood). 2001;226:1051-60.
11 Powell JM, Sonnenfeld G.The effects of dehydroepiandrosterone (DHEA) on in vitro spleen cell proliferation and cytokine production. J Interferon Cytokine Res. 2006;26:34-9.
12 Du C, Khalil MW, Sriram S. Administration of dehydro-epiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice.J Immunol. 2001; 167: 7094-101.
13 Yang BC, Liu CW, Chen YC, et al. Exogenous dehydro-epiandrosterone modified the expression of T helper-related cytokines in NZB/NZW F1 mice.Immunol Invest. 1998;27:291-302.
14 van Vollenhoven RF, Morabito LM, Engleman EG, et al.Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J Rheumatol. 1998;25:285-9.
15 van Vollenhoven RF. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3:23-31.
16 Straub RH, Scholmerich J, Zietz B.Replacement therapy with DHEA plus corticosteroids in patients with chronic inflammatory diseases-substitutes of adrenal and sex hormones.Z Rheumatol. 2000;59 Suppl 2:11/108-18.
17 Maurer M,Gold R.Animal models of immune-mediated neuropathies. Curr Opin Neurol. 2002;15:617-22.
1 Maurer M, Gold R. Animal models of immune-mediated neuropathies. Curr Opin Neurol. 2002 ;15:617-22.
2 Gold R, Hartung HP, Toyka KV.Animal models for autoimmune demyelinating disorders of the nervous system.Mol Med Today. 2000; 6:88-91.
3 Norton WT, Poduslo SE.Myelination in rat brain: method of myelin isolation. J Neurochem. 1973 ;21:749-57. FEBS Lett.
4 Kitamura K, Suzuki M, Suzuki A,et al.The complete amino acid sequence of the P2 protein in bovine peripheral nerve myelin. FEBS Lett. 1980 Jun 16; 115:27-30.
5 Uyemura K, Suzuki M, Kitamura K, et al. Neuritogenic determinant of bovine P2 protein in peripheral nerve myelin. J Neurochem. 1982;39:895-8.
6 Kuwabara S. Guillain-barre syndrome. Curr Neurol Neurosci Rep. 2007; 7: 57-62.
7 Hughes RA, Cornblath DR.Guillain-Barre syndrome. Lancet. 2005; 366: 1653-66.
8 Waksman BH, Adams RD. Allergic neuritis: an experimental disease of rabbits induced by the injection of peripheral nervous tissue and adjuvants.J Exp Med. 1955; 102:213-36.
9 Milner P, Lovelidge CA, Taylor WA,et al.PO myelin protein produces experimental allergic neuritis in Lewis rats. J Neurol Sci. 1987;79:275-85.
10 Ben-Nun A, Wekerle H, Cohen IR.The rapid isolation of clonable antigen-specific T lymphocyte lines capable of mediating autoimmune encephalomyelitis.Eur J Immunol. 1981 ;11:195-9.
11 Olee T, Powell HC, Brostoff SW. New minimum length requirement for a T cell epitope for experimental allergic neuritis. J Neuroimmunol. 1990; 27: 187-90
12 Taylor WA, Hughes RA.Experimental allergic neuritis induced in SJL mice by bovine P2.J Neuroimmunol. 1985; 8:153-7.
13 Calida DM, Kremlev SG, Fujioka T,et al.Experimental allergic neuritis in the SJL/J mouse: induction of severe and reproducible disease with bovine peripheral nerve myelin and pertussis toxin with or without interleukin-12. J Neuroimmunol. 2000;107:l-7.
14 Zou LP, Ljunggren HG, Levi M, et al.PO protein peptide 180-199 together with pertussis toxin induces experimental autoimmune neuritis in resistant C57BL/6 mice.J Neurosci Res. 2000;62:717-21.
15 Zhu J, Zou L, Zhu S,et al.Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) blockade enhances incidence and severity of experimental autoimmune neuritis in resistant mice.J Neuroimmunol. 2001;115:111-7.
1 Miller RA. Aging and immune function.Int Rev Cytol. 1991; 124: 187-215.
2 Svec F, Porter JR.The actions of exogenous dehydroepiandrosterone in experimental animals and humans.Proc Soc Exp Biol Med. 1998 ;218:174-91.
3 Oberbeck R, Dahlweid M, Koch R,et al. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29:380-4.
4 Small M, Gray CE, Beastall GH, et al. Adrenal androgens in insulin-dependent diabetes mellitus, Diabetes Res. 1989;11:93-5.
5 Henderson E, Schwartz A, Pashko L,et al. Dehydroepiandrosterone and 16 alpha-bromo-epiandrosterone: inhibitors of Epstein-Barr virus- induced transformation of human lymphocytes. Carcinogenesis. 1981; 2:683-6.
6 Khaw KT. Dehydroepiandrosterone, dehydroepiandrosterone sulphate and cardiovascular disease.J Endocrinol. 1996 ;150 Suppl: S149-53.
7 Du C, Khalil MW, Sriram S.Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice.J Immunol. 2001; 167 :7094-101.
8 Rontzsch A, Thoss K, Petrow PK,et al. Amelioration of murine antigen- induced arthritis by dehydroepiandrosterone (DHEA). Inflamm Res. 2004;53:189-98.
9 van Vollenhoven RF. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002;3: 23-31.
10 Dillon JS. Dehydroepiandrosterone, Dehydroepiandrosterone Sulfate and Related Steroids: Their Role in Inflammatory, Allergic and Immunological Disorders. Curr Drug Targets Inflamm Allergy. 2005;4:377-85.
11 Stephanie JW, Thomas EG, Russell AP. The biological action of Dehydroepiandrosterone involves multiple receptors. Drug Metabolism. 2006;38: 89-116.
12 von Mühlen D, Laughlin GA, Kritz-Silverstein D, et al. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods. Contemp Clin Trials. 2007;28:153-68.
13 Dessein PH, Joffe BI, Stanwix AE, et al.Hyposecretion of the adrenal androgen dehydroepiandrosterone sulfate and its relation to clinical variables in inflammatory arthritis.Arthritis Res. 2001 ;3:183-8.
14 Khalkhali-Ellis Z, Moore TL, Hendrix MJ.Reduced levels of testosterone and dehydroepiandrosterone sulphate in the serum and synovial fluid of juvenile rheumatoid arthritis patients correlates with disease severity.Clin Exp Rheumatol. 1998;16:753-6.
15 Straub RH, Zeuner M, Antoniou E, et al. Dehydroepiandrosterone sulfate is positively correlated with soluble interleukin 2 receptor and soluble intercellular adhesion molecule in systemic lupus erythematosus.J Rheumatol. 1996;23:856-61.
16 de la Torre B, von Krogh G, Svensson M, et al. Blood cortisol and dehydroepiandrosterone sulphate (DHEAS) levels and CD4 T cell counts in HIV infection. Clin Exp Rheumatol. 1997;15:87-90.
17 Henderson E, Yang JY, Schwartz A Dehydroepiandrosterone (DHEA) and synthetic DHEA analogs are modest inhibitors of HIV-1 IIIB replication. AIDS Res Hum Retroviruses. 1992; 8:625-31.
18 van Vollenhoven RF, Morabito LM, Engleman EG, et al. Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J Rheumatol. 1998;25:285-9.
19 van Vollenhoven RF. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3: 23-31.
20 Svec F, Porter JR.The actions of exogenous dehydroepiandrosterone in experimental animals and humans.Proc Soc Exp Biol Med. 1998; 218: 174- 91.
21 Oberbeck R, Dahlweid M, Koch R,et al. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29:380-4.
22 Padgett DA, Loria RM, Sheridan JF. Steroid hormone regulation of antiviral immunity. Ann N Y Acad Sci. 2000;917:935-43.
23 Loria RM. Immune up-regulation and tumor apoptosis by androstene steroids. Steroids. 2002 ; 7: 53-66.
24 Loria RM, Conrad DH, Huff T,et al. Androstenetriol and androstenediol. Protection against lethal radiation and restoration of immunity after radiation injury.Ann N Y Acad Sci. 2000;917:860-7.
25 Du C, Khalil MW, Sriram S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice.J Immunol. 2001;167: 7094-101.
26 Rontzsch A, Thoss K, Petrow PK, et al. Amelioration of murine antigen- induced arthritis by dehydroepiandrosterone (DHEA). Inflamm Res. 2004; 53: 189-98.
27 Offner H, Zamora A, Drought H, et al.A synthetic androstene derivative and a natural androstene metabolite inhibit relapsing-remitting EAE. J Neuroimmunol. 2002;130:128-39.
28 Offner H, Zamora A, Subramanian S, et al. A synthetic androstene analogue inhibits collagen-induced arthritis in the mouse. Clinical Immunology. 2004;l 10:181-90.
29 Yang BC, Liu CW, Chen YC, et al.Exogenous dehydroepiandrosterone modified the expression of T helper-related cytokines in NZB/NZW Fl mice.Immunol Invest. 1998;27:291-302.
30 Duan, R.S., Link, H., Xiao, B.G. Dehydroepiandrosterone therapy ameliorates experimental autoimmune myasthenia gravis in Lewis rats. J Clin Immunol. 2003;23:100-6.
31 Yoneda M, Wada K, Katayama K, et al. A novel therapy for acute hepatitis utilizing dehydroepiandrosterone in the murine model of hepatitis. Biochem Pharmacol. 2004;68:2283-9.
32 Auci D, Nicoletti F, Mangano K, et al.Anti-inflammatory and Immune Regulatory Properties of 5-Androsten-3_, 17_-Diol (HE2100), and Synthetic Analogue HE3204 Implications for Treatment of Autoimmune Diseases. Ann. N.Y. Acad. Sci. 2005;1051: 730-742.
33 Kipper-Galperin M, Galilly R, Danenberg HD, et al. Dehydroepiandrosterone selectively inhibits production of tumor necrosis factor-a and interlukin-6 in astrocytes. Int. J. Dev. Neurosci. 1999;17:765-75.
1 Hughes RA, Cornblath DR.Guillain-Barre syndrome. Lancet. 2005; 366:1653-66.
2 Kuwabara S. Guillain-barre syndrome. Curr Neurol Neurosci Rep. 2007; 7: 57-62.
3 Svec F, Porter JR.The actions of exogenous dehydroepiandrosterone in experimental animals and humans.Proc Soc Exp Biol Med. 1998; 218: 174- 91.
4 von Mühlen D, Laughlin GA, Kritz-Silverstein D, et al. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods[J]. Contemp Clin Trials, 2007,28:153-168.
5 Straub RH, Vogl D, Gross V,et al. Association of humoral markers of inflammation and dehydroepiandrosterone sulfate or cortisol serum levels in patients with chronic inflammatory bowel disease.Am J Gastroenterol. 1998; 93:2197-202.
6 Straub RH, Paimela L, Peltomaa R, et L.Inadequately low serum levels of steroid hormones in relation to interleukin-6 and tumor necrosis factor in untreated patients with early rheumatoid arthritis and reactive arthritis.Arthritis Rheum. 2002;46:654-62.
7 Straub RH, Lehle K, Herfarth H, et al.Dehydroepiandrosterone in relation to other adrenal hormones during an acute inflammatory stressful disease state compared with chronic inflammatory disease: role of interleukin-6 and tumour necrosis factor.Eur J Endocrinol. 2002;146:365-74.
8 Suzuki T, Suzuki N, Engleman EG, et al. Low serum levels of dehydroepiandrosterone may cause deficient IL-2 production by lymphocytes in patients with systemic lupus erythematosus (SLE).Clin Exp Immunol. 1995; 99:251-5.
9 Straub RH, Zeuner M, Antoniou E, et al. Dehydroepiandrosterone sulfate is positively correlated with soluble interleukin 2 receptor and soluble intercellular adhesion molecule in systemic lupus erythematosus.J Rheumatol. 1996;23:856-61.
10 Simard J, Gingras S. Crucial role of cytokines in sex steroid formation in normal and tumoral tissues.Mol Cell Endocrinol. 2001;171:25-40.
11 Turgeon C, Gingras S, Carriere MC, et al. Regulation of sex steroid formation by nterleukin-4 and interleukin-6 in breast cancer cells.J Steroid Biochem Mol Biol. 1998;65:151-62.
12 Tabata N, Tagami H, Terui T. Dehydroepiandrosterone may be one of the regulators of cytokine production in atopic dermatitis.Arch Dermatol Res. 1997;289:410-4.
13 Hernandez-Pando R, De La Luz Streber M, Orozco H, et al. The effects of androstenediol and dehydroepiandrosterone on the course and cytokine profile of tuberculosis in BALB/c mice.Immunology. 1998;95:234-41.
14 Suzuki T, Suzuki N, Daynes RA, et al. Dehydroepiandrosterone enhances IL2 production and cytotoxic effector function of human T cells.Clin Immunol Immunopathol. 1991 ;61:202-11.
15 Canning MO, Grotenhuis K, de Wit HJ, et al.Opposing effects of dehydroepiandrosterone and dexamethasone on the generation of monocyte-derived dendritic cells. Eur J Endocrinol. 2000; 143: 687-95.
16 Zhang Z, Araghi-Niknam M, Liang B, et al.Prevention of immune dysfunction and vitamin E loss by dehydroepiandrosterone and melatonin supplementation during murine retrovirus infection. Immunology. 1999;96:291-7
17 Yu CK, Yang BC, Lei HY,et al. Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response. Clin Exp Allergy. 1999;29:414-22
18 Du C, Guan Q, Khalil MW, et al.Stimulation of Th2 response by high doses of dehydroepiandrosterone in KLH-primed splenocytes.Exp Biol Med (Maywood). 2001 Dec;226:1051-60.
19 Powell JM, Sonnenfeld G.The effects of dehydroepiandrosterone (DHEA) on in vitro spleen cell proliferation and cytokine production. J Interferon Cytokine Res. 2006;26:34-9.
20 Du C, Khalil MW, Sriram S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice.J Immunol. 2001 ;167:7094-101.
21 Yang BC, Liu CW, Chen YC, et al . Exogenous dehydroepiandrosterone modified the expression of T helper-related cytokines in NZB/NZW F1 mice.
Immunol Invest. 1998;27(4-5):291-302.
22 Di Santo E, Sironi M, Mennini T, et al. A glucocorticoid receptor-independent mechanism for neurosteroid inhibition of tumor necrosis factor production.Eur J Pharmacol. 1996;299:179-86.
23 Rontzsch A, Thoss K, Petrow PK, Henzgen S, Brauer R. Amelioration of murine antigen-induced arthritis by dehydroepiandrosterone (DHEA). Inflamm Res. 2004;53:189-98.
24 Offner H, Zamora A, Drought H, et al.A synthetic androstene derivative and a natural androstene metabolite inhibit relapsing-remitting EAE.J Neuroimmunol. 2002; 130:128-39.
25 Offner H, Zamora A, Subramanian S, et al. A synthetic androstene analogue inhibits collagen-induced arthritis in the mouse. Clin Immunol. 2004; 110: 181-90.
26 van Vollenhoven RF, Morabito LM, Engleman EG, et al.Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J Rheumatol. 1998;25:285-9.
27 van Vollenhoven RF. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3: 23-31.
28 Giltay EJ, van Schaardenburg D, Gooren LJ, et al. Dehydro-
epiandrosterone sulfate in patients with rheumatoid arthritis.Ann N Y Acad Sci. 1999;876:152-4.
29 Labrie F, Luu-The V, Labrie C, et al.DHEA and its transformation into androgens and estrogens in peripheral target tissues: intracrinology.Front Neuroendocrinol. 2001;22:185-212.
30 Sugiyama H, Yamada J, Takama H, et al.Photoaffinity labeling of peroxisome proliferator binding proteins in rat hepatocytes; dehydroepiandrosterone sulfate- and bezafibrate-binding proteins. Biochim Biophys Acta. 1997; 1339: 321-30.
31 Tsuji K, Furutama D, Tagami M, et al.Specific binding and effects of dehydroepiandrosterone sulfate (DHEA-S) on skeletal muscle cells: possible implication for DHEA-S replacement therapy in patients with myotonic dystrophy.Life Sci. 1999;65:17-26.
32 Bruder JM, Sobek L, Oettel M. Dehydroepiandrosterone stimulates the estrogen response element. J Steroid Biochem Mol Biol. 1997;62:461-6.
33 Tan J, Sharief Y, Hamil KG, et al.Dehydroepiandrosterone activates mutant androgen receptors expressed in the androgen-dependent human prostate cancer xenograft CWR22 and LNCaP cells.Mol Endocrinol. 1997;11(4): 450-9.
34 Peters JM, Zhou YC, Ram PA,et al. Peroxisome proliferator-activated receptor alpha required for gene induction by dehydroepiandrosterone- 3beta-sulfate. Mol Pharmacol. 1996;50:67-74.
35 Poynter ME, Daynes RA. Peroxisome proliferator-activated receptor alpha activation modulates cellular redox status, represses nuclear factor-kappaB signaling, and reduces inflammatory cytokine production in aging. J Biol Chem. 1998;273:32833-41.
36 Aragno M, Parola S, Tamagno E, et al. Oxidative derangement in rat synaptosomes induced by hyperglycaemia: restorative effect of dehydroepiandrosterone treatment.Biochem Pharmacol. 2000 Aug l;60:389-95.
37 Zhu Y, Ljunggren HG, Mix E, et al. Suppression of autoimmune neuritis in IFN-gamma receptor-deficient mice. Experimental Neurology. 2001; 169: 472-478.
38 Weishaupt A, Bruck W, Hartung T, et al. Schwann cell apoptosis in experimental autoimmune neuritis of the Lewis rat and the functional role of tumor necrosis factor-alpha. Neurosci Lett, 2001, 306: 77-80.
39 Dillon JS. Dehydroepiandrosterone, dehydroepiandrosterone sulfate and related steroids: their role in inflammatory, allergic and immunological disorders. Curr Drug Targets Inflamm Allergy, 2005,4:377-385.
1 Maurer M., Gold R. Animal models of immune-mediated neuropathies. Curr Opin Neurol. 2002; 15, 617-622.
2 Hughes R.A., Cornblath, D.R(?) Guillain-Barre syndrome. Lancet. 2005; 366, 1653-1666.
3 Kuwabara S., Guillain-barre syndrome. Curr Neurol Neurosci Rep. 2007;7, 57-62.
4 von Muhlen D., Laughlin G.A., Kritz-Silverstein D., et al. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods. Contemp Clin Trials. 2007;28, 153-168.
5 Svec F., Porter J.R. The actions of exogenous dehydroepiandrosterone in experimental animals and humans. Proc Soc Exp Biol Med. 1998;218, 174-191.
6 Oberbeck R., Dahlweid M., Koch R.,.et al. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29, 380-384.
7 Yu C.K., Yang B.C., Lei H.Y., et al. Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response. Clin Exp Allergy. 1999;29, 414-22.
8 Du C., Guan Q., Khalil M.W., et al. Stimulation of Th2 response by high doses of dehydroepiandrosterone in KLH-primed splenocytes. Exp Biol Med (Maywood). 2001;226, 1051-1060.
9 Powell J.M., Sonnenfeld G. The effects of dehydroepiandrosterone (DHEA) on in vitro spleen cell proliferation and cytokine production. J Interferon Cytokine Res. 2006;26, 34-39.
10 Du C., Khalil M.W., Sriram S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice. J Immunol. 2001;167, 7094-7101.
11 Yang B.C., Liu C.W., Chen Y.C., et al. Exogenous dehydroepiandrosterone modified the expression of T helper-related cytokines in NZB/NZW F1 mice. Immunol Invest. 1998;27, 291-302.
12 Straub R.H., Scholmerich J., Zietz B. Replacement therapy with DHEA plus corticosteroids in patients with chronic inflammatory diseases--substitutes of adrenal and sex hormones. Z. Rheumatol. 2000; 59 Suppl 2:II, 108-118.
13 van Vollenhoven R.F. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3, 23-31.
14 van Vollenhoven R.F., Morabito L.M., Engleman E.G., et al. Treatment of systemic lupus erythematosus with dehydro- epiandrosterone: 50 patients treated up to 12 months. J. Rheumatol. 1998; 25, 285-289.
15 Norton W.T., Poduslo S.E. Myelination in rat brain: method of myelin isolation. J. Neurochem. 1973; 21,749-757.
16 Auci D., Nicoletti F., Mangano K.,et al. Anti-inflammatory and Immune Regulatory Properties of 5-Androsten-3_β 17_β-Diol (HE2100), and Synthetic Analogue HE3204 Implications for Treatment of Autoimmune Diseases. Ann. N.Y. Acad. Sci. 2005;1051, 730-742.
17 Duan R.S., Link H., Xiao B.G. Dehydroepiandrosterone therapy ameliorates experimental autoimmune myasthenia gravis in Lewis rats. J Clin Immunol. 2003;23, 100-106.
18 Offner H., Zamora A., Drought H., et al. A synthetic androstene derivative and a natural androstene metabolite inhibit relapsing-remitting EAE. J. Neuroimmunol. 2002;30, 128-139.
19 Offner H., Zamora A., Subramanian S., et al. synthetic androstene analogue inhibits collagen-induced arthritis in the mouse. Clin Immunol. 2004; 110, 181-190.
20 Rontzsch A., Thoss K., Petrow P.K., et al. Amelioration of murine antigen-induced arthritis by dehydroepiandrosterone (DHEA). Inflamm Res. 2004;53, 189-198.
21 Yoneda M., Wada K., Katayama K., et al. A novel therapy for acute hepatitis utilizing dehydroepiandrosterone in the murine model of hepatitis. Biochem Pharmacol. 2004;68,2283-2289.
22 Kipper-Galperin M., Galilly R., Danenberg H.D., et al. Dehydroepiandrosterone selectively inhibits production of tumor necrosis factor-a and interlukin-6 in astrocytes. Int. J. Dev. Neurosci. 1999;17, 765-775.
23 Zhu, J., Mix, E., Link, H. Cytokine production and the pathogenesis of experimental autoimmune neuritis and Guillain-Barre syndrome. J. Neuroimmunol. 1998; 84, 40-52.
24 Zhu Y., Ljunggren H.G., Mix E., et al. Suppression of autoimmune neuritis in IFN-gamma receptor-deficient mice. Exp Neurol. 2001;169, 472-478.
25 Redford E.J., Hall S.M., Smith K.J. Vascular changes and demyelination induced by the intraneural injection of tumour necrosis factor. Brain. 1995;118, 869-878.
26 Weishaupt A., Bruck W., Hartung T., et al. Schwann cell apoptosis in experimental autoimmune neuritis of the Lewis rat and the functional role of tumor necrosis factor-alpha. Neurosci Lett. 2001; 306, 77-80.
27 Elkarim, R.A., Dahle, C., Mustafa, M.,et al. Recovery from Guillain-Barre syndrome is associated with increased levels of neutralizing autoantibodies to interferon-gamma. Clin Immunol Immunopathol. 1998;88, 241-248.
28 Hohnoki, K., Inoue, A., Koh, C.S. Elevated serum levels of IFN-gamma, IL-4 and TNF-alpha/unelevated serum levels of IL-10 in patients with demyelinating diseases during the acute stage. J. Neuroimmunol. 1998; 87, 27-32.
29 Misawa, S., Kuwabara, S., Mori, M., et al. Serum levels of tumor necrosis factor-alpha in chronic inflammatory demyelinating polyneuropathy. Neurology. 2001; 56, 666-669.
30 Radhakrishnan, V.V., Sumi, M.G., Reuben, S., et al. Serum tumour necrosis factor-alpha and solubletumour necrosis factor receptors levels in patients withGuillain-Barre syndrome. Acta Neurol Scand . 2004;109, 71-74.
31 Sharief ,M.K., Ingram, D.A., Swash, M. Circulating tumor necrosis factor-alpha correlates with electrodiagnostic abnormalities in Guillain-Barre syndrome. Ann Neurol. 1997;42, 68-73.
32 Dillon,J.S. Dehydroepiandrosterone, Dehydroepiandrosterone Sulfate and Related Steroids: Their Role in Inflammatory, Allergic and Immunological Disorders. Curr Drug Targets Inflamm Allergy. 2005; 4,377-385.
1 Svec, F., Porter, J.R. The actions of exogenous dehydroepiandrosterone in experimental animals and humans. Proc Soc Exp Biol Med. 1998; 218, 174-191.
2. von Mühlen, D., Laughlin, G.A., Kritz-Silverstein, D., Barrett-Connor, E. The Dehydroepiandrosterone And WellNess (DAWN) study: research design and methods. Contemp Clin Trials. 2007; 28, 153-168.
3 Oberbeck, R., Dahlweid, M., Koch, R., van Griensven, M., Emmendorfer, A., Tscherne, H., Pape, H.C. Dehydroepiandrosterone decreases mortality rate and improves cellular immune function during polymicrobial sepsis. Crit Care Med. 2001;29, 380-384.
4 Yu, C.K., Yang, B.C., Lei, H.Y., Chen, Y.C., Liu, Y.H., Chen, C.C., Liu, C.W. Attenuation of house dust mite Dermatophagoides farinae-induced airway allergic responses in mice by dehydroepiandrosterone is correlated with down-regulation of TH2 response. Clin Exp Allergy. 1999; 29, 414-22.
5 Du, C., Guan, Q., Khalil M.W., Sriram, S. Stimulation of Th2 response by high doses of dehydroepiandrosterone in KLH-primed splenocytes. Exp Biol Med (Maywood). 2001; 226, 1051-1060.
6 Straub, R.H., Scholmerich, J., Zietz, B. Replacement therapy with DHEA plus corticosteroids in patients with chronic inflammatory diseases—substitutes of adrenal and sex hormones. Z. Rheumatol. 2000;59 Suppl 2:11, 108-118.
7 van Vollenhoven, R.F. Dehydroepiandrosterone for the treatment of systemic lupus erythematosus. Expert Opin Pharmacother. 2002; 3, 23-31.
8 van Vollenhoven, R.F., Morabito, L.M., Engleman, E.G., McGuire, J.L.Treatment of systemic lupus erythematosus with dehydroepiandrosterone: 50 patients treated up to 12 months. J. Rheumatol. 1998;25, 285-289.
9 Maurer M, Gold R.Animal models of immune-mediated neuropathies. Curr Opin Neurol. 2002 ;15:617-22.
10 Gold R, Hartung HP, Toyka KV. Animal models for autoimmune demyelinating disorders of the nervous system. Mol Med Today. 2000; 6: 88-91.
11 Norton, W.T., Poduslo, S.E. Myelination in rat brain: method of myelin isolation. J. Neurochem. 1973 ;21,749-757.
12 Du C, Khalil MW, Sriram S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice. J Immunol. 2001,167:7094-7101.
13 Duan RS, Link H, Xiao BG. Dehydroepiandrosterone therapy ameliorates experimental autoimmune myasthenia gravis in Lewis rats. J Clin Immunol. 2003,23(2):100-106.
14 Shin HC, McFarlane EF, Pollard JD, Watson EG. Induction of experimental allergic neuritis with synthetic peptides from myelin P2 protein. Neurosci Lett 1989;102:309-12
15 Linington C, Izumo S, Suzuki M, Uyemura K, Meyermann R,Wekerle H. A permanent rat T cell line that mediates experimental allergic neuritis in the Lewis rat in vivo. J Immunol 1984; 133:1946-50
16 Linington C, Lassmann H, Ozawa K, Kosin S, Mongan L. Cell adhesion molecules of the immunoglobulin supergene family as tissuespecific autoantigens: Induction of experimental allergic neuritis (EAN) by P0 protein-specific T cell lines. Eur J Immunol 1992;22:1813-17
17 Hahn AF. Experimental allergic neuritis (EAN) as a model for the immune-mediated demyelinating neuropathies. Rev Neurol 1996;152:328—32
18 Du, C., Khalil, M.W., Sriram, S. Administration of dehydroepiandrosterone suppresses experimental allergic encephalomyelitis in SJL/J mice. J Immunol. 2001;167, 7094-7101.
19 Offner, H., Zamora, A., Drought, H., Matejuk, A., Auci, D.L., Morgan, E.E., Vandenbark, A.A., Reading, C.L. A synthetic androstene derivative and a natural androstene metabolite inhibit relapsing-remitting EAE. J. Neuroimmunol. 2002;30, 128-139.
20 Zhu, J., Mix, E., Link, H. Cytokine production and the pathogenesis of experimental autoimmune neuritis and Guillain-Barre syndrome. J. Neuroimmunol. 1998;84,40-52.
21 Zhu, Y., Ljunggren, H.G., Mix, E., Li, H.L., van der Meide, P., Elhassan, A.M., Winblad, B., Zhu, J. Suppression of autoimmune neuritis in IFN-gamma receptor-deficient mice. Exp Neurol. 2001;169, 472-478.
22 Redford, E.J., Hall, S.M., Smith, K.J. Vascular changes and demyelination induced by the intraneural injection of tumour necrosis factor. Brain. 1995; 118,869-878.
23 Weishaupt ,A., Bruck, W., Hartung, T., Toyka, K.V., Gold, R. Schwann cell apoptosis in experimental autoimmune neuritis of the Lewis rat and the functional role of tumor necrosis factor-alpha. Neurosci Lett. 2001;306, 77-80.
24 Dillon, J.S. Dehydroepiandrosterone, Dehydroepiandrosterone Sulfate and Related Steroids: Their Role in Inflammatory, Allergic and Immunological Disorders. Curr Drug Targets Inflamm Allergy. 2005; 4, 377-385.
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