全血灌流免疫吸附治疗实验性变态反应性神经炎的研究
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摘要
目的:探讨全血灌流免疫吸附治疗实验性变态反应性神经炎的效果和作用机制。
方法:分别应用髓鞘碱性蛋白(MBP)和牛坐骨神经免疫新西兰兔,从中选择适合全血灌流的动物模型。健康兔随机分为4组,分别为实验组1组(接受牛坐骨神经免疫,在免疫后第14天给予1次全血灌流免疫吸附治疗),实验组2组(接受牛坐骨神经免疫,在免疫后第14天和第16天分别给予1次全血灌流免疫吸附治疗),阳性对照组(接受牛坐骨神经免疫,但不给予任何治疗处理)和阴性对照组(给予生理盐水5 ml多点皮内注射,在免疫后14天接受1次全血灌流)。应用Graham K临床评分对实验兔进行临床评定。阴性对照组实验兔在免疫后第14天进行全血灌流,通过检测灌流前后血细胞的变化进行血液相容性的评价,在免疫后第18天同阳性对照组实验兔一同进行运动神经传导速度、波幅和F波测定,两组还需在免疫后第35天取坐骨神经进行髓鞘染色,切片进行光镜检查。实验组1组和实验组2组在免疫后第14天进行灌流治疗,两天后实验组2组进行第2次灌流治疗,灌流前后取血进行血清成分分析,在免疫后第35天两组兔进行神经电生理检测和病理观察。定期留取4组实验兔血清并测量其血清P2抗体的滴度变化。应用木瓜蛋白酶和胃蛋白酶水解健康兔IgG,水解片段由吸附剂进行吸附,并比较各水解片段的吸附率。
结果:用MBP和坐骨神经均可制备新西兰兔EAN,后者更适合全血灌流操作。阳性对照组兔自免疫后14.13±0.64天出现临床症状,在免疫后第28～32天症状达到高峰,自免疫后35.00±0.63天存活兔开始恢复,总病程约60.80±0.84天。实验组1组实验兔在免疫后第32.13±0.64天开始恢复,总病程为42.38±0.50天。实验组2组实验兔在免疫后第30.25±0.46天开始恢复,总病程为38.38±0.52天。坐骨神经病理变化在阳性对照组实验兔病情高峰期表现为髓鞘脱失严重,轴索有损伤。经灌流治疗后,神经脱髓鞘现象明显减轻,这种改善在实验组2组表现的最为显著。阳性对照组在免疫后第18天电生理检查示神经传导速度轻度降低,在免疫后第35天进展到重度减慢;波幅和F波出现率为中度减低。经灌流治疗后,神经传导速度改善到中度减低,波幅和F波出现率转变为正常。坐骨神经免疫组实验兔在免疫后第5天即可检测到血清P2抗体,其血清浓度在阳性对照组总体上经历了一个先是急剧上升然后逐渐下降的曲线变化。在接受灌流治疗后,曲线变得较为平缓,曲线的这种变化在实验组2组体现地更为明显。阴性对照组实验兔灌流前后的血常规、血脂及电解质无显著性变化(p＞0.05)。灌流治疗对血清白蛋白无明显影响,但可降低血清球蛋白浓度。酶水解IgG可得F(ab')2、Fc和Fab,吸附率分别为17.94±4.10%、2.48±2.63%和0.02±0.05%。
结论:球形纤维素为载体色氨酸为配基的吸附剂血液相容性良好,对IgG具有非特异性的吸附作用,作用靶点可能为IgG的铰链区。同阳性对照组相比,全血灌流免疫吸附治疗可使免疫兔在病理、电生理和血清P2抗体浓度的检测上呈现出改善的趋势。基于此,可认为全血灌流免疫吸附治疗对EAN兔具有积极的治疗作用,且治疗效果同灌流次数有一定关系,这或许可为GBS提供新的治疗策略。
Objective:Evaluate the therapy of extracorporeal whole blood immunoadsorption on experimental allergic neuritis and access the mechanism of cellulose-tryptophan on the adsorption.
Methods:For optimize the immunogen of EAN rabbits,myelin basic protein (MBP)was extracted from bovine sciatic nerve.The New Zealand rabbit EAN model was induced by intradermic injection of MBP or bovine sciatic nerve.The healthy rabbits were divided into four groups,i.e.experimental group one(treated with immunoadsorption for one time at the day 14 after immunization),experimental group two(underwent immunoadsorption twice,at the day 14 and the day 16 after immunization),positive control group(EAN rabbits without any treatment)and negative control group(physiologic saline solution instead of MBP and underwent immunoadsorption at the same time with group two).According to Graham K,the clinical manifestation of the EAN rabbits was graded in 7 levels.The blood samples were taken from the rabbits before and after therapy for the assessment of the biocompatibility.To the negative and positive control groups,the electrophysiological functions,including motor nerve conduction velocity(MNCV),motor evoked potential amplitude,F wave were evaluated at the day 18 after immunization.All the rabbits were comprehensively examined,e.g.electrophysiological function and pathological changes,at the day 35 after immunonization.ELISA was introduced for the measurement of serum P2 protein antibody.The blood samples for the detection of P2 protein antibody were regularly drawn from all the rabbits,from the day 0 to the day 60 after immunization.F(ab')~2,Fab,and Fc fragments of IgG from the health rabbits were achieved by pepsin and papain,for the further investigation on the mechanism of cellulose-tryptophan.
Results:EAN was successfully induced either by MBP or sciatic nerve.EAN rabbits set up by the sciatic nerve undertook the extracorporeal whole blood immunoadsorption.In positive control group,symptoms climbed to the climax from the day 28 to the day 32,remitted from the day 35.00±0.63,and the mean duration was 60.80±0.84 days.The rabbits in the experimental group one developed the neurological signs of EAN at the day 14.25±0.46,ameliorated since the day 32.13± 0.64,the duration is 42.38±0.50.The experimental group two,rabbits recovered at the day 30.25±0.46,the duration is 38.38±0.52.The perivascular infiltration of macrophages and demyelination in the sciatic nerve were found in positive control group.Experimental group showed lighter pathological changes,especially in experimental group two.The positive control group,at the day 18,MNCV retarded obviously,amplitude fell down,the detection rate of F wave reduced and the electrophysiological function continued to deteriorate until the day 35.Compared with positive control group,the electrophysiological function in the experiment groups significant improved(p＜0.05).Anti-P2 IgG could be detected at the day 5 post-inoculation and plasma levels showed a sharply rise and gradual fell down throughout the experimental period in positive control group.No significant changes of blood cells,electrolytes and blood lipids were found.Serum albumin was not altered by adsorbent,but the concentration of globulin dropped down.The adsorption rate of F(ab')~2 and Fc was 17.94±4.10%and 2.48±2.63%,no adsorption was observed with Fab.
Conclusion:Imunoadsorbent with cellulose as carrier tryptophan as ligand was biocompatible with the blood system and had specific affinity with Globulin.It was inferred that cellulose-tryptophan most likely bond with the hinge region of IgG. Compared with positive control,experimental group displayed less severe clinical sign,faster recovery of electrophysiological and pathological characteristics of sciatic nerves.Based on these results,it can be concluded that extracorporeal whole blood immunoadsorption has therapeutical effect on EAN rabbits.
方法:分别应用髓鞘碱性蛋白(MBP)和牛坐骨神经免疫新西兰兔,从中选择适合全血灌流的动物模型。健康兔随机分为4组,分别为实验组1组(接受牛坐骨神经免疫,在免疫后第14天给予1次全血灌流免疫吸附治疗),实验组2组(接受牛坐骨神经免疫,在免疫后第14天和第16天分别给予1次全血灌流免疫吸附治疗),阳性对照组(接受牛坐骨神经免疫,但不给予任何治疗处理)和阴性对照组(给予生理盐水5 ml多点皮内注射,在免疫后14天接受1次全血灌流)。应用Graham K临床评分对实验兔进行临床评定。阴性对照组实验兔在免疫后第14天进行全血灌流,通过检测灌流前后血细胞的变化进行血液相容性的评价,在免疫后第18天同阳性对照组实验兔一同进行运动神经传导速度、波幅和F波测定,两组还需在免疫后第35天取坐骨神经进行髓鞘染色,切片进行光镜检查。实验组1组和实验组2组在免疫后第14天进行灌流治疗,两天后实验组2组进行第2次灌流治疗,灌流前后取血进行血清成分分析,在免疫后第35天两组兔进行神经电生理检测和病理观察。定期留取4组实验兔血清并测量其血清P2抗体的滴度变化。应用木瓜蛋白酶和胃蛋白酶水解健康兔IgG,水解片段由吸附剂进行吸附,并比较各水解片段的吸附率。
结果:用MBP和坐骨神经均可制备新西兰兔EAN,后者更适合全血灌流操作。阳性对照组兔自免疫后14.13±0.64天出现临床症状,在免疫后第28～32天症状达到高峰,自免疫后35.00±0.63天存活兔开始恢复,总病程约60.80±0.84天。实验组1组实验兔在免疫后第32.13±0.64天开始恢复,总病程为42.38±0.50天。实验组2组实验兔在免疫后第30.25±0.46天开始恢复,总病程为38.38±0.52天。坐骨神经病理变化在阳性对照组实验兔病情高峰期表现为髓鞘脱失严重,轴索有损伤。经灌流治疗后,神经脱髓鞘现象明显减轻,这种改善在实验组2组表现的最为显著。阳性对照组在免疫后第18天电生理检查示神经传导速度轻度降低,在免疫后第35天进展到重度减慢;波幅和F波出现率为中度减低。经灌流治疗后,神经传导速度改善到中度减低,波幅和F波出现率转变为正常。坐骨神经免疫组实验兔在免疫后第5天即可检测到血清P2抗体,其血清浓度在阳性对照组总体上经历了一个先是急剧上升然后逐渐下降的曲线变化。在接受灌流治疗后,曲线变得较为平缓,曲线的这种变化在实验组2组体现地更为明显。阴性对照组实验兔灌流前后的血常规、血脂及电解质无显著性变化(p＞0.05)。灌流治疗对血清白蛋白无明显影响,但可降低血清球蛋白浓度。酶水解IgG可得F(ab')2、Fc和Fab,吸附率分别为17.94±4.10%、2.48±2.63%和0.02±0.05%。
结论:球形纤维素为载体色氨酸为配基的吸附剂血液相容性良好,对IgG具有非特异性的吸附作用,作用靶点可能为IgG的铰链区。同阳性对照组相比,全血灌流免疫吸附治疗可使免疫兔在病理、电生理和血清P2抗体浓度的检测上呈现出改善的趋势。基于此,可认为全血灌流免疫吸附治疗对EAN兔具有积极的治疗作用,且治疗效果同灌流次数有一定关系,这或许可为GBS提供新的治疗策略。
Objective:Evaluate the therapy of extracorporeal whole blood immunoadsorption on experimental allergic neuritis and access the mechanism of cellulose-tryptophan on the adsorption.
Methods:For optimize the immunogen of EAN rabbits,myelin basic protein (MBP)was extracted from bovine sciatic nerve.The New Zealand rabbit EAN model was induced by intradermic injection of MBP or bovine sciatic nerve.The healthy rabbits were divided into four groups,i.e.experimental group one(treated with immunoadsorption for one time at the day 14 after immunization),experimental group two(underwent immunoadsorption twice,at the day 14 and the day 16 after immunization),positive control group(EAN rabbits without any treatment)and negative control group(physiologic saline solution instead of MBP and underwent immunoadsorption at the same time with group two).According to Graham K,the clinical manifestation of the EAN rabbits was graded in 7 levels.The blood samples were taken from the rabbits before and after therapy for the assessment of the biocompatibility.To the negative and positive control groups,the electrophysiological functions,including motor nerve conduction velocity(MNCV),motor evoked potential amplitude,F wave were evaluated at the day 18 after immunization.All the rabbits were comprehensively examined,e.g.electrophysiological function and pathological changes,at the day 35 after immunonization.ELISA was introduced for the measurement of serum P2 protein antibody.The blood samples for the detection of P2 protein antibody were regularly drawn from all the rabbits,from the day 0 to the day 60 after immunization.F(ab')~2,Fab,and Fc fragments of IgG from the health rabbits were achieved by pepsin and papain,for the further investigation on the mechanism of cellulose-tryptophan.
Results:EAN was successfully induced either by MBP or sciatic nerve.EAN rabbits set up by the sciatic nerve undertook the extracorporeal whole blood immunoadsorption.In positive control group,symptoms climbed to the climax from the day 28 to the day 32,remitted from the day 35.00±0.63,and the mean duration was 60.80±0.84 days.The rabbits in the experimental group one developed the neurological signs of EAN at the day 14.25±0.46,ameliorated since the day 32.13± 0.64,the duration is 42.38±0.50.The experimental group two,rabbits recovered at the day 30.25±0.46,the duration is 38.38±0.52.The perivascular infiltration of macrophages and demyelination in the sciatic nerve were found in positive control group.Experimental group showed lighter pathological changes,especially in experimental group two.The positive control group,at the day 18,MNCV retarded obviously,amplitude fell down,the detection rate of F wave reduced and the electrophysiological function continued to deteriorate until the day 35.Compared with positive control group,the electrophysiological function in the experiment groups significant improved(p＜0.05).Anti-P2 IgG could be detected at the day 5 post-inoculation and plasma levels showed a sharply rise and gradual fell down throughout the experimental period in positive control group.No significant changes of blood cells,electrolytes and blood lipids were found.Serum albumin was not altered by adsorbent,but the concentration of globulin dropped down.The adsorption rate of F(ab')~2 and Fc was 17.94±4.10%and 2.48±2.63%,no adsorption was observed with Fab.
Conclusion:Imunoadsorbent with cellulose as carrier tryptophan as ligand was biocompatible with the blood system and had specific affinity with Globulin.It was inferred that cellulose-tryptophan most likely bond with the hinge region of IgG. Compared with positive control,experimental group displayed less severe clinical sign,faster recovery of electrophysiological and pathological characteristics of sciatic nerves.Based on these results,it can be concluded that extracorporeal whole blood immunoadsorption has therapeutical effect on EAN rabbits.
引文
[1]陈可冀.格林—巴利综合征[M].北京:中国医药科技出旅社,2000:2.
[2]Zhang Z,Zhang ZY,Fauser U,et al.FTY720 ameliorates experimental autoimmune neuritis by inhibition of lymphocyte and monocyte infiltration into peripheral nerves[J].Exp Neurol,2008,210(2):681-690.
[3]Maurer M,Toyka KV,Gold R.Cellular immunity in inflammatory autoimmune neuropathies[J].Rev Neurol(Paris),2002,158(12):7-15.
[4]Zhu J,Mix E,Link H.Cytokine production and the pathogenesis of experimental autoimmune neuritis and Guillain-Barre syndrome[J].J Neuroimmunol,1998,84(1):40-52.
[5]Creange A.A Role for Interferon-beta in Guillain-Barre Syndrome?[J].BioDrugs,2000,14(1):1-11.
[6]Yamazaki Z,Fujimori Y,Takahama T,et al.Efficiency and biocompatibility of a new immunosorbent[J].Trans Am Soc Artif Intern Organs,1982,28(3):318-323.
[7]Yang KS,Kenpe K,Yamaji K,et al.Plasma adsorption in critical care[J].Ther Apher,2002,6(3):184-188.
[8]Okamiya S,Ogino M,Ogino Y,et al.Tryptophan-immobilized column-based immunoadsorption as the choice method for plasmapheresis in Guillain-Barre syndrome[J].Ther Apher Dial,2004,8(3):248-253.
[9]Latov N.Practice parameter:immunotherapy for Guillain-Barre syndrome:report of the Quality Standards Subcommittee of the American Academy of Neurology[J].Neurology,2004,62(9):1653-1654.
[10]Yang L,Cheng Y,Yah WR,et al.Extracorporeal whole blood immunoadsorption of autoimmune myasthenia gravis by cellulose tryptophan adsorbent[J].Artif Cells Blood Substit Immobil Biotechnol,2004,32(4):519-528.
[11]WAKSMAN BH,ADAMS RD.Allergic neuritis:an experimental disease of rabbits induced by the injection of peripheral nervous tissue and adjuvants[J].J Exp Med,1955,102(2):213-236.
[12]London Y.Ox peripheral nerve myelin membrane.Purification and partial characterization of two basic proteins[J].Biochim Biophys Acta,1971,249(1):188-196.
[13]吴抒见,陈娟,朱高发,等.豚鼠实验性变态反应性神经炎模型的建立及电生理、免疫等指标的观察[J].中华神经科杂志,1995,28(6):66-69.
[14]李建武,萧能,余瑞元,等.生物化学实验原理与方法[M].北京:北京大学出版社,1994:168-170,216-223.
[15]贾怡,陈文捷,李虹,等.牛MBP诱导BALB/c和C57BL/6小鼠实验性变态性脑脊髓炎样反应的研究[J].现代免疫学杂志,2005,25(3):187-190.
[16]王惠,杨天祝,林嘉友.髓鞘素的提取与鉴定[J].河北医科大学学报,2001,22(1):31-32.
[17]张旭,许贤豪.牛坐骨神经髓鞘碱性蛋白提取及初步应用[J].温州医学院学报,1993,23(4):215-218.
[18]Harvey GK,Schindhelm K,Antony JH,et al.Linear response curves from an ELISA assay:measurement of anti-myelin IgG and IgM during experimental allergic neuritis[J].J Neurosci Methods,1987,21(1):81-90.
[19]高长玉,张国华,王桂荣登.格林—巴利综合征的电生理分型研究[J].中华物理医学与康复杂志,1995,17(1):25-28.
[20]Zhang ZY,Zhang Z,Fauser U,et al.Improved outcome of EAN,an animal model of GBS,through amelioration of peripheral and central inflammation by minocycline[J].J Cell Mol Med,2008,8:2-5.
[21]Stienekemeier M,Falk K,Rotzschke O,et al.Vaccination,prevention,and treatment of experimental autoimmune neuritis(EAN)by an oligomerized T cell epitope[J].Proc Natl Acad Sci U S A,2001,98(24):13872-13877.
[22]Jung S,Gaupp S,Hartung HP,et al.Oral tolerance in experimental autoimmune neuritis(EAN)of the Lewis rat.Ⅱ.Adjuvant effects and bystander suppression in P2 peptide-induced EAN[J].J Neuroimmunol,2001,116(1):21-28.
[23]吴云,王化冰,王维治.Th1/Th2型细胞因子在实验性自身免疫性神经炎发病机制中的作用[J].中华神经科杂志,2007,40(9):626-629.
[24]Jung S,Gaupp S,Hartung HP,et al.Oral tolerance in experimental autoimmune neuritis(EAN)of the Lewis rat.Ⅱ.Adjuvant effects and bystander suppression in P2 peptide-induced EAN[J].J Neuroimmunol,2001,116(1):21-28.
[25]王翠娣,郭玉璞.实验性变态反应性神经炎动物模型的建立和病理特点[J].中华神经精神科杂志,1995,28(4):202-205.
[26]王琳,张凤蕴.MBP的提取、鉴定及EAE模型的建立[J].哈尔滨医科大学学 报,1998,32(1):12-13.
[27]郭向东,朱锡华.P2蛋白诱导兔慢性变态反应性神经炎的研究[J].免疫学杂志,1998,14(1):28-31.
[28]李勇,郑荣远,李剑敏,等.实验性变态反应性脑脊髓炎大鼠模型的复制[J].中国病理生理杂志,2005,21(2):414-416.
[29]Hahn AF,Feasby TE,Steele A,et al.Demyelination and axonal degeneration in Lewis rat experimental allergic neuritis depend on the myelin dosage[J].Lab Invest,1988,59(1):115-125.
[30]Abramsky O,London Y.Purification and partial characterization of two basic proteins from human peripheral nerve[J].Biochim Biophys Acta,1975,393(2):556-562.
[31]郭向东,朱锡华.髓鞘碱性蛋白Ⅱ的纯化与鉴定[J].免疫学杂志,1997,13(4):239-242.
[32]Cavaletti G,Mata S,Fasano A,et al.Lipid-free versus lipid-bound P2protein-induced experimental allergic neuritis;clinicopathological,neurophysiological,and immunological study[J].J Neurosci Res,2000:62(5):709-716.
[33]杨丽,程焱.全血灌流吸附治疗重症肌无力的实验研究[J].中华急诊医学杂志,2004,13(10):670-672.
[34]胡玉红,恽时锋,周森妹等.速眠新麻醉对家兔的生理影响[J].中国比较医学杂志,2006,16(8):475-478.
[35]文小玲,张平,姜德诵.速眠新与戊巴比妥钠在动物麻醉中的应用比较[J].南华大学学报:医学版,2004,3(1):131-132.
[36]刘豫安,夏叶玲,主编.基础医学功能实验教程[M].北京:人民卫生出版社,2000:71.
[37]姜冠华.体外循环中的肺保护[J].山东医药,2005,45(6):62-63.
[38]Hartung HP,Schafer B,Heininger K,et al.The role of macrophages and eicosanoids in the pathogenesis of experimental allergic neuritis.Serial clinical,electrophysiological,biochemical and morphological observations[J].Brain,1988,111(5):1039-1059.
[39]白蓉主编.实验动物学基础及技术[M].北京:人民卫生出版社,2003:58.
[40]陈主初,吴端生主编.实验动物学[M].长沙:湖南科学技术出版社,2001:188.
[41]孔令字,刘晓波,朱振宇等.抗体F(ab′)2片段制备的新方法-木瓜蛋白酶酶切法[J].临床检验杂志,2004,22(2):115-116.
[42]刘智广,陈志南,刘成刚等.FPLC纯化单克隆抗体(Fab′)-2片段[J].细胞与分子免疫学杂志,1994,10(3):51-56.
[43]Andrew SM,Titus JA.Fragmentation of immunoglobulin G[J].Curr Protoc Immunol,2001,Chapter 2:Unit 2.8.
[44]Li H,Lockyer S,Concepcion A,et al.The Fab fragment of a novel anti-GPVI monoclonal antibody,OM4,reduces in vivo thrombosis without bleeding risk in rats[J].Arterioscler Thromb Vase Biol,2007,27(5):1199-1205.
[45]咸海青,范明.重组人睫状神经营养因子的复性研究[J].生物化学与生物物理进展,1999,26(5):454-457.
[46]Clifford C J,Downes S,A comparative study of the use of colorimtric assays in the assessment of biocompatibility[J].J Mater Sci Master Medicine,1996,7(10):637-643.
[47]张志友,张文怡,钱绍诚.犬肝动脉输注阿霉素联合血液灌流的研究[J].世界华人消化杂志,2003,11(8):1160-1163.
[48]马昌义,彭克军,杜一平,等.炭肾清除血液中破伤风毒素效能的实验探讨[J].中华肾脏病杂志,2005,21(12):751-752.
[49]翟翚,杨丽.全血灌流吸附治疗动物实验方法的建立[J].天津医科大学学报,2004,10(1):65-67.
[50]何中心,郭兴.颈外静脉插管至上腔静脉的应用解剖[J].医学理论与实践,2002,15(4):383-384.
[51]陈友燕,叶斌.颈外静脉穿刺应用解剖及临床意义[J].中华护理杂志,2000,35(3):160-162.
[52]南开大学实验动物解剖学编写组编.实验动物解剖学[M].北京:高等教育出版社出版,1979:23-32.
[53]郝洁.血液灌流技术治疗急性中毒的应用及护理[J].中国现代医药杂志,2004,6(6):66-67.
[54]刘晓莉,刘均敏,刘小君.经颈外静脉插管至深静脉行血液透析的效果观察[J].南方护理学报,2002,9(3):8-9.
[55]于美丽,贾克培,吕文岚等.血液灌流清除体内免疫复合物的实验研究[J].中 国生物医学工程学报,2000,20(1):12-16.
[56]关广聚,时一民主编.临床血液净化学[M].济南:山东科学技术出版社,2003:213-222.
[57]Gabriel CM,Hughes RA,Moore SE,et al.Induction of experimental autoimmune neuritis with peripheral myelin protein-22[J].Brain,1998,121(Pt 10):1895-1902.
[58]Taylor JM,Pollard JD.Neurophysiological changes in demyelinating and axonal forms of acute experimental autoimmune neuritis in the Lewis rat[J].Muscle Nerve,2003,28(3):344-352.
[59]田玉旺,丁华野,李琳等.介绍一种神经髓鞘染色法[J].临床与实验病理学杂志,2003,19(4):436-436.
[60]田玉旺,李琳,李丽等.神经髓鞘染色新方法的建立及其原理探讨[J].中国组织化学与细胞化学杂志,2004,13(4):543-545.
[61]Bechtold DA,Yue X,Evans RM,et al.Axonal protection in experimental autoimmune neuritis by the sodium channel blocking agent flecainide[J].Brain,2005,128(1):18-28.
[62]Gold R,Hartung HP,Toyka KV.Animal models for autoimmune demyelinating disorders of the nervous system[J].Mol Med Today,2000,6(2):88-91.
[63]周红,乔健,吕传真等.吉兰.巴雷综合征患者血清和脑脊液中P2蛋白抗体检测及其临床意义[J].中国神经精神疾病杂志,2000,26(5):296-297.
[64]陈俊杰;王若菡;李昌隆等.简易的髓鞘碱性蛋白及其抗体酶联免疫吸附同步定量测定法[J].华西医科大学学报,1995,26(2):125-130.
[65]郭向东,许贤豪,李成文.兔、豚鼠抗牛P2多克隆抗血清的制备与鉴定[J].中国神经免疫学和神经病学杂志,1997,4(4):243.
[66]邓国民,林嘉友,许贤豪.格林-巴利综合征患者脑脊液抗P2蛋白IgG抗体增高[J].中国神经免疫学和神经病学杂志,1994,1(2):98-102.
[67]米力,陈志南,余晓玲等.单克隆抗体F(ab′)_2片段的制备与质量控制[J].中国生物制品学杂志,1999,12(4)216-218.
[68]孔宪涛主编.免疫球蛋白异常的基础和临床[M].上海:上海科学技术文献出版社,1997:9.
1.Willison HJ,Townson K,Veitch J,et al.Synthetic disialylgalactose immunoadsorbents deplete anti-GQ1b antibodies from autoimmune neuropathy sera[J].Brain,2004,127(3):680-691.
2.Finsterer J,Treatment of immune-mediated,dysimmune neuropathies[J].Acta Neurol Scand,2005,112(2):115-125.
3.Chaudhuri A.Guillain-Barre syndrom[J]e.Lancet,2006,367(9509):472-473.
4.Komagamine T,Yuki N.Ganglioside mimicry as a cause of Guillain-Barre syndrome[J].CNS Neurol Disord Drug Targets,2006,5(4):391-400.
5.Overell JR,Willison HJ,Recent developments in Miller Fisher syndrome and related disorders[J].Curr Opin Neurol,2005,18(5):562-566.
6.Yamazaki Z,Fujimori Y,Takahama T,et al.Efficiency and biocompatibility of a new immunosorbent[J].Trans Am Soc Artif Intern Organs,1982,28:318-323.
7.Jimenez C,Rosenow F,Grieb P,et al.Adsorption therapy with tryptophan-conjugated polyvinyl alcohol gels in 10 patients with acute Guillain-Barre syndrome[J].Transfus Sci,1993,14(1):9-11.
8.Yang KS,Kenpe K,Yamaji K,et al.Plasma adsorption in critical care[J].Ther Apher,2002,6(3):184-188.
9.Takei H,Komaba Y,Araki T,et al.Plasma immunoadsorption therapy for Guillain-Barre syndrome:critical day for initiation[J].J Nippon Med Sch,2002,69(6):557-563.
10.Okamiya S,Ogino M,Ogino Y,et al.Tryptophan-immobilized column-based immunoadsorption as the choice method for plasmapheresis in Guillain-Barre syndrome[J].Ther Apher Dial,2004,8(3):248-253.
11.Latov N.Practice parameter:immunotherapy for Guillain-Barre syndrome:report of the Quality Standards Subcommittee of the American Academy of Neurology[J]. Neurology, 2004, 62(9): 1653-1654.
12. Harel M, Shoenfeld Y.Intravenous immunoglobulin and Guillain-Barre syndrome[J]. Clin Rev Allergy Immunol, 2005, 29(3):281-287.
13. Haupt WF, Birkmann C, Pawlik G, et al. Apheresis and selective adsorption plus immunoglobulin treatment in Guillain-Barre syndrome[J]. Ther Apher, 2000, 4(3): 198-200. .
14. Rajabally YA, Strens LH, Abbott RJ. Acute motor conduction block neuropathy followed by axonal degeneration and poor recovery[J]. Neurology, 2006, 66(2):287-288.
15. Seta T, Nagayama H, Katsura K, et al. Factors influencing outcome in Guillain-Barre Syndrome: comparison of plasma adsorption against other treatments[J]. Clin Neurol Neurosurg, 2005, 107(6):491-496.
16. Koyama K, Dei F, Koyano S, et al. The therapeutic efficacy in Fisher syndrome[J]. No To Shinkei, 2005, 57(2): 149-152.
17. Shinoda T, Arakura H, Yamaguchi E, et al. Immunoadsorption plasmapheresis and plasma exchange in Guillain-Barre syndrome [J]. Ther Plasmapheresis, 1993, 12(2):531-534.
18. Yuki N. Tryptophan-immobilized column adsorbs immunoglobulin G anti-GQ1b antibody from Fisher's syndrome: A new approach to treatment[J]. Neurology, 1996, 46(6): 1644-1651.
19. Shahar E. Current therapeutic options in severe Guillain-Barre syndrome[J]. Clin Neuropharmacol, 2006,29(1):45-51.
20. van Doom PA. Treatment of Guillain-Barre syndrome and CIDP[J]. J Peripher Nerv Syst, 2005, 10(2): 113-127.
21. Stegmayr BG. A survey of blood purification techniques [J]. Transfus Apher Sci, 2005, 32(2):209-220.
22. Yang L, Cheng Y, Yan WR, et al. Extracorporeal whole blood immunoadsorption of autoimmune myasthenia gravis by cellulose tryptophan adsorbent[J]. Artif Cells Blood Substit Immobil Biotechnol, 2004, 32(4):519-528.
[2]Zhang Z,Zhang ZY,Fauser U,et al.FTY720 ameliorates experimental autoimmune neuritis by inhibition of lymphocyte and monocyte infiltration into peripheral nerves[J].Exp Neurol,2008,210(2):681-690.
[3]Maurer M,Toyka KV,Gold R.Cellular immunity in inflammatory autoimmune neuropathies[J].Rev Neurol(Paris),2002,158(12):7-15.
[4]Zhu J,Mix E,Link H.Cytokine production and the pathogenesis of experimental autoimmune neuritis and Guillain-Barre syndrome[J].J Neuroimmunol,1998,84(1):40-52.
[5]Creange A.A Role for Interferon-beta in Guillain-Barre Syndrome?[J].BioDrugs,2000,14(1):1-11.
[6]Yamazaki Z,Fujimori Y,Takahama T,et al.Efficiency and biocompatibility of a new immunosorbent[J].Trans Am Soc Artif Intern Organs,1982,28(3):318-323.
[7]Yang KS,Kenpe K,Yamaji K,et al.Plasma adsorption in critical care[J].Ther Apher,2002,6(3):184-188.
[8]Okamiya S,Ogino M,Ogino Y,et al.Tryptophan-immobilized column-based immunoadsorption as the choice method for plasmapheresis in Guillain-Barre syndrome[J].Ther Apher Dial,2004,8(3):248-253.
[9]Latov N.Practice parameter:immunotherapy for Guillain-Barre syndrome:report of the Quality Standards Subcommittee of the American Academy of Neurology[J].Neurology,2004,62(9):1653-1654.
[10]Yang L,Cheng Y,Yah WR,et al.Extracorporeal whole blood immunoadsorption of autoimmune myasthenia gravis by cellulose tryptophan adsorbent[J].Artif Cells Blood Substit Immobil Biotechnol,2004,32(4):519-528.
[11]WAKSMAN BH,ADAMS RD.Allergic neuritis:an experimental disease of rabbits induced by the injection of peripheral nervous tissue and adjuvants[J].J Exp Med,1955,102(2):213-236.
[12]London Y.Ox peripheral nerve myelin membrane.Purification and partial characterization of two basic proteins[J].Biochim Biophys Acta,1971,249(1):188-196.
[13]吴抒见,陈娟,朱高发,等.豚鼠实验性变态反应性神经炎模型的建立及电生理、免疫等指标的观察[J].中华神经科杂志,1995,28(6):66-69.
[14]李建武,萧能,余瑞元,等.生物化学实验原理与方法[M].北京:北京大学出版社,1994:168-170,216-223.
[15]贾怡,陈文捷,李虹,等.牛MBP诱导BALB/c和C57BL/6小鼠实验性变态性脑脊髓炎样反应的研究[J].现代免疫学杂志,2005,25(3):187-190.
[16]王惠,杨天祝,林嘉友.髓鞘素的提取与鉴定[J].河北医科大学学报,2001,22(1):31-32.
[17]张旭,许贤豪.牛坐骨神经髓鞘碱性蛋白提取及初步应用[J].温州医学院学报,1993,23(4):215-218.
[18]Harvey GK,Schindhelm K,Antony JH,et al.Linear response curves from an ELISA assay:measurement of anti-myelin IgG and IgM during experimental allergic neuritis[J].J Neurosci Methods,1987,21(1):81-90.
[19]高长玉,张国华,王桂荣登.格林—巴利综合征的电生理分型研究[J].中华物理医学与康复杂志,1995,17(1):25-28.
[20]Zhang ZY,Zhang Z,Fauser U,et al.Improved outcome of EAN,an animal model of GBS,through amelioration of peripheral and central inflammation by minocycline[J].J Cell Mol Med,2008,8:2-5.
[21]Stienekemeier M,Falk K,Rotzschke O,et al.Vaccination,prevention,and treatment of experimental autoimmune neuritis(EAN)by an oligomerized T cell epitope[J].Proc Natl Acad Sci U S A,2001,98(24):13872-13877.
[22]Jung S,Gaupp S,Hartung HP,et al.Oral tolerance in experimental autoimmune neuritis(EAN)of the Lewis rat.Ⅱ.Adjuvant effects and bystander suppression in P2 peptide-induced EAN[J].J Neuroimmunol,2001,116(1):21-28.
[23]吴云,王化冰,王维治.Th1/Th2型细胞因子在实验性自身免疫性神经炎发病机制中的作用[J].中华神经科杂志,2007,40(9):626-629.
[24]Jung S,Gaupp S,Hartung HP,et al.Oral tolerance in experimental autoimmune neuritis(EAN)of the Lewis rat.Ⅱ.Adjuvant effects and bystander suppression in P2 peptide-induced EAN[J].J Neuroimmunol,2001,116(1):21-28.
[25]王翠娣,郭玉璞.实验性变态反应性神经炎动物模型的建立和病理特点[J].中华神经精神科杂志,1995,28(4):202-205.
[26]王琳,张凤蕴.MBP的提取、鉴定及EAE模型的建立[J].哈尔滨医科大学学 报,1998,32(1):12-13.
[27]郭向东,朱锡华.P2蛋白诱导兔慢性变态反应性神经炎的研究[J].免疫学杂志,1998,14(1):28-31.
[28]李勇,郑荣远,李剑敏,等.实验性变态反应性脑脊髓炎大鼠模型的复制[J].中国病理生理杂志,2005,21(2):414-416.
[29]Hahn AF,Feasby TE,Steele A,et al.Demyelination and axonal degeneration in Lewis rat experimental allergic neuritis depend on the myelin dosage[J].Lab Invest,1988,59(1):115-125.
[30]Abramsky O,London Y.Purification and partial characterization of two basic proteins from human peripheral nerve[J].Biochim Biophys Acta,1975,393(2):556-562.
[31]郭向东,朱锡华.髓鞘碱性蛋白Ⅱ的纯化与鉴定[J].免疫学杂志,1997,13(4):239-242.
[32]Cavaletti G,Mata S,Fasano A,et al.Lipid-free versus lipid-bound P2protein-induced experimental allergic neuritis;clinicopathological,neurophysiological,and immunological study[J].J Neurosci Res,2000:62(5):709-716.
[33]杨丽,程焱.全血灌流吸附治疗重症肌无力的实验研究[J].中华急诊医学杂志,2004,13(10):670-672.
[34]胡玉红,恽时锋,周森妹等.速眠新麻醉对家兔的生理影响[J].中国比较医学杂志,2006,16(8):475-478.
[35]文小玲,张平,姜德诵.速眠新与戊巴比妥钠在动物麻醉中的应用比较[J].南华大学学报:医学版,2004,3(1):131-132.
[36]刘豫安,夏叶玲,主编.基础医学功能实验教程[M].北京:人民卫生出版社,2000:71.
[37]姜冠华.体外循环中的肺保护[J].山东医药,2005,45(6):62-63.
[38]Hartung HP,Schafer B,Heininger K,et al.The role of macrophages and eicosanoids in the pathogenesis of experimental allergic neuritis.Serial clinical,electrophysiological,biochemical and morphological observations[J].Brain,1988,111(5):1039-1059.
[39]白蓉主编.实验动物学基础及技术[M].北京:人民卫生出版社,2003:58.
[40]陈主初,吴端生主编.实验动物学[M].长沙:湖南科学技术出版社,2001:188.
[41]孔令字,刘晓波,朱振宇等.抗体F(ab′)2片段制备的新方法-木瓜蛋白酶酶切法[J].临床检验杂志,2004,22(2):115-116.
[42]刘智广,陈志南,刘成刚等.FPLC纯化单克隆抗体(Fab′)-2片段[J].细胞与分子免疫学杂志,1994,10(3):51-56.
[43]Andrew SM,Titus JA.Fragmentation of immunoglobulin G[J].Curr Protoc Immunol,2001,Chapter 2:Unit 2.8.
[44]Li H,Lockyer S,Concepcion A,et al.The Fab fragment of a novel anti-GPVI monoclonal antibody,OM4,reduces in vivo thrombosis without bleeding risk in rats[J].Arterioscler Thromb Vase Biol,2007,27(5):1199-1205.
[45]咸海青,范明.重组人睫状神经营养因子的复性研究[J].生物化学与生物物理进展,1999,26(5):454-457.
[46]Clifford C J,Downes S,A comparative study of the use of colorimtric assays in the assessment of biocompatibility[J].J Mater Sci Master Medicine,1996,7(10):637-643.
[47]张志友,张文怡,钱绍诚.犬肝动脉输注阿霉素联合血液灌流的研究[J].世界华人消化杂志,2003,11(8):1160-1163.
[48]马昌义,彭克军,杜一平,等.炭肾清除血液中破伤风毒素效能的实验探讨[J].中华肾脏病杂志,2005,21(12):751-752.
[49]翟翚,杨丽.全血灌流吸附治疗动物实验方法的建立[J].天津医科大学学报,2004,10(1):65-67.
[50]何中心,郭兴.颈外静脉插管至上腔静脉的应用解剖[J].医学理论与实践,2002,15(4):383-384.
[51]陈友燕,叶斌.颈外静脉穿刺应用解剖及临床意义[J].中华护理杂志,2000,35(3):160-162.
[52]南开大学实验动物解剖学编写组编.实验动物解剖学[M].北京:高等教育出版社出版,1979:23-32.
[53]郝洁.血液灌流技术治疗急性中毒的应用及护理[J].中国现代医药杂志,2004,6(6):66-67.
[54]刘晓莉,刘均敏,刘小君.经颈外静脉插管至深静脉行血液透析的效果观察[J].南方护理学报,2002,9(3):8-9.
[55]于美丽,贾克培,吕文岚等.血液灌流清除体内免疫复合物的实验研究[J].中 国生物医学工程学报,2000,20(1):12-16.
[56]关广聚,时一民主编.临床血液净化学[M].济南:山东科学技术出版社,2003:213-222.
[57]Gabriel CM,Hughes RA,Moore SE,et al.Induction of experimental autoimmune neuritis with peripheral myelin protein-22[J].Brain,1998,121(Pt 10):1895-1902.
[58]Taylor JM,Pollard JD.Neurophysiological changes in demyelinating and axonal forms of acute experimental autoimmune neuritis in the Lewis rat[J].Muscle Nerve,2003,28(3):344-352.
[59]田玉旺,丁华野,李琳等.介绍一种神经髓鞘染色法[J].临床与实验病理学杂志,2003,19(4):436-436.
[60]田玉旺,李琳,李丽等.神经髓鞘染色新方法的建立及其原理探讨[J].中国组织化学与细胞化学杂志,2004,13(4):543-545.
[61]Bechtold DA,Yue X,Evans RM,et al.Axonal protection in experimental autoimmune neuritis by the sodium channel blocking agent flecainide[J].Brain,2005,128(1):18-28.
[62]Gold R,Hartung HP,Toyka KV.Animal models for autoimmune demyelinating disorders of the nervous system[J].Mol Med Today,2000,6(2):88-91.
[63]周红,乔健,吕传真等.吉兰.巴雷综合征患者血清和脑脊液中P2蛋白抗体检测及其临床意义[J].中国神经精神疾病杂志,2000,26(5):296-297.
[64]陈俊杰;王若菡;李昌隆等.简易的髓鞘碱性蛋白及其抗体酶联免疫吸附同步定量测定法[J].华西医科大学学报,1995,26(2):125-130.
[65]郭向东,许贤豪,李成文.兔、豚鼠抗牛P2多克隆抗血清的制备与鉴定[J].中国神经免疫学和神经病学杂志,1997,4(4):243.
[66]邓国民,林嘉友,许贤豪.格林-巴利综合征患者脑脊液抗P2蛋白IgG抗体增高[J].中国神经免疫学和神经病学杂志,1994,1(2):98-102.
[67]米力,陈志南,余晓玲等.单克隆抗体F(ab′)_2片段的制备与质量控制[J].中国生物制品学杂志,1999,12(4)216-218.
[68]孔宪涛主编.免疫球蛋白异常的基础和临床[M].上海:上海科学技术文献出版社,1997:9.
1.Willison HJ,Townson K,Veitch J,et al.Synthetic disialylgalactose immunoadsorbents deplete anti-GQ1b antibodies from autoimmune neuropathy sera[J].Brain,2004,127(3):680-691.
2.Finsterer J,Treatment of immune-mediated,dysimmune neuropathies[J].Acta Neurol Scand,2005,112(2):115-125.
3.Chaudhuri A.Guillain-Barre syndrom[J]e.Lancet,2006,367(9509):472-473.
4.Komagamine T,Yuki N.Ganglioside mimicry as a cause of Guillain-Barre syndrome[J].CNS Neurol Disord Drug Targets,2006,5(4):391-400.
5.Overell JR,Willison HJ,Recent developments in Miller Fisher syndrome and related disorders[J].Curr Opin Neurol,2005,18(5):562-566.
6.Yamazaki Z,Fujimori Y,Takahama T,et al.Efficiency and biocompatibility of a new immunosorbent[J].Trans Am Soc Artif Intern Organs,1982,28:318-323.
7.Jimenez C,Rosenow F,Grieb P,et al.Adsorption therapy with tryptophan-conjugated polyvinyl alcohol gels in 10 patients with acute Guillain-Barre syndrome[J].Transfus Sci,1993,14(1):9-11.
8.Yang KS,Kenpe K,Yamaji K,et al.Plasma adsorption in critical care[J].Ther Apher,2002,6(3):184-188.
9.Takei H,Komaba Y,Araki T,et al.Plasma immunoadsorption therapy for Guillain-Barre syndrome:critical day for initiation[J].J Nippon Med Sch,2002,69(6):557-563.
10.Okamiya S,Ogino M,Ogino Y,et al.Tryptophan-immobilized column-based immunoadsorption as the choice method for plasmapheresis in Guillain-Barre syndrome[J].Ther Apher Dial,2004,8(3):248-253.
11.Latov N.Practice parameter:immunotherapy for Guillain-Barre syndrome:report of the Quality Standards Subcommittee of the American Academy of Neurology[J]. Neurology, 2004, 62(9): 1653-1654.
12. Harel M, Shoenfeld Y.Intravenous immunoglobulin and Guillain-Barre syndrome[J]. Clin Rev Allergy Immunol, 2005, 29(3):281-287.
13. Haupt WF, Birkmann C, Pawlik G, et al. Apheresis and selective adsorption plus immunoglobulin treatment in Guillain-Barre syndrome[J]. Ther Apher, 2000, 4(3): 198-200. .
14. Rajabally YA, Strens LH, Abbott RJ. Acute motor conduction block neuropathy followed by axonal degeneration and poor recovery[J]. Neurology, 2006, 66(2):287-288.
15. Seta T, Nagayama H, Katsura K, et al. Factors influencing outcome in Guillain-Barre Syndrome: comparison of plasma adsorption against other treatments[J]. Clin Neurol Neurosurg, 2005, 107(6):491-496.
16. Koyama K, Dei F, Koyano S, et al. The therapeutic efficacy in Fisher syndrome[J]. No To Shinkei, 2005, 57(2): 149-152.
17. Shinoda T, Arakura H, Yamaguchi E, et al. Immunoadsorption plasmapheresis and plasma exchange in Guillain-Barre syndrome [J]. Ther Plasmapheresis, 1993, 12(2):531-534.
18. Yuki N. Tryptophan-immobilized column adsorbs immunoglobulin G anti-GQ1b antibody from Fisher's syndrome: A new approach to treatment[J]. Neurology, 1996, 46(6): 1644-1651.
19. Shahar E. Current therapeutic options in severe Guillain-Barre syndrome[J]. Clin Neuropharmacol, 2006,29(1):45-51.
20. van Doom PA. Treatment of Guillain-Barre syndrome and CIDP[J]. J Peripher Nerv Syst, 2005, 10(2): 113-127.
21. Stegmayr BG. A survey of blood purification techniques [J]. Transfus Apher Sci, 2005, 32(2):209-220.
22. Yang L, Cheng Y, Yan WR, et al. Extracorporeal whole blood immunoadsorption of autoimmune myasthenia gravis by cellulose tryptophan adsorbent[J]. Artif Cells Blood Substit Immobil Biotechnol, 2004, 32(4):519-528.