未成熟树突状细胞诱导实验性自身免疫性重症肌无力耐受的试验研究  

Experimental study on the induction of tolerance to experimental autoimmune myasthenia gravis by immature dendritic cells

作　　者：李罗清[1] 孙圣刚[1] 曹学兵[1] 王云甫[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院神经科,武汉430022

出　　处：《中华微生物学和免疫学杂志》2005年第4期263-268,共6页Chinese Journal of Microbiology and Immunology

基　　金：华中科技大学同济医学院课题资助 (No :2 0 0 3 16) ;湖北省教育厅重点资助 (No :2 0 0 3A0 0 6)

摘　　要：目的　探讨未成熟树突状细胞(iDC)对实验性自身免疫性重症肌无力(EAMG)的影响。方法　用小剂量粒 巨噬细胞集落刺激因子(GM -CSF)体外诱导骨髓前体细胞,获得的iDC用乙酰胆碱受体(AChR)负载后皮下接种同基因大鼠,3周后通过体外试验观察脾细胞对AChR的增殖反应,同时用AChR和完全弗氏佐剂(CFA)免疫大鼠,观察免疫-7周后MG相关指标的改变。结果　小剂量GM -CSF诱导所获得的iDC与成熟DC(mDC)相比,其表面MHCⅡ和共刺激分子CD80、CD86的表达低,摄取dextran -FITC的能力强但刺激同种T细胞增殖的能力弱。接种iDC、mDC、AChR负载的mDC的大鼠和对照组一样,体外脾细胞对AChR的刺激均强烈增殖,用AChR和CFA免疫后,产生明显的MG症状,重复电刺激出现明显衰减,血清AChRAb滴度明显增高,神经肌肉接头呈现典型的MG样改变;而接种AChR负载的iDC的大鼠脾细胞对AChR的刺激呈现明显的增殖抑制但对卵清蛋白的刺激仍强烈增殖,MG相关指标均未见明显改变。结论　骨髓前体细胞经小剂量GM -CSF诱导产生了iDC ,AChR负载的iDC可诱导EAMG耐受。Objective To investigate the effect of immature dendritic cells(iDCs) on experimental autoimmune myasthenia gravis(EAMG). Methods iDCs were induced in low dose of GM-CSF from bone marrow precursors of Lewis rats and determined the phenotype and function, then the iDCs were pulsed with acetylcholine receptor(AChR) and transferred to allogeneic rats. The proliferative response of spleen cells to AChR in vitro study was viewed after 3 weeks, then all rats were immunized with AChR and complete Freund′s adjuvant(CFA), then viewed for the corresponding indexes of MG for 7 weeks. Results Compared with mature DCs(mDCs) induced in high dose of GM-CSF with the additional stimulation by lipopolysaccharide, iDCs expressed significantly lower levels of MHCⅡ, CD80 and CD86, their capabilities of uptaking dextran-FITC were stronger but proliferation of allogeneic T cell were inhibited. As controls, spleen cells of all rats transferred iDCs, mDCs and AChR pulsed mDCs proliferated markedly to AChR in vitro study. After immunization, they showed typical clinical symptoms in 4 to 7 weeks. Their electromyogram wave amplitude attenuated obviously, serum AChRAb increased and neuromuscular junction appeared as typical damage of MG. In contrast, proliferative response of spleen cells of rats transferred AChR pulsed iDCs was dramatically inhibited to AChR but proliferated strongly to ovalbumin. There was no manifest signs in these rats after immunization. Conclusion iDCs could be generated by inducing bone marrow precursors in low dose of GM-CSF, AChR pulsed iDCs could induce tolerance of EAMG.

关 键 词：实验性自身免疫性重症肌无力 试验研究 细胞诱导 粒-巨噬细胞集落刺激因子 耐受 未成熟树突状细胞 骨髓前体细胞 乙酰胆碱受体 完全弗氏佐剂 ACHRAB 神经肌肉接头 GM-CSF iDC GMCSF 共刺激分子 T细胞增殖 重复电刺激 小剂量 

分 类 号：R746.1[医药卫生—神经病学与精神病学] R730.5[医药卫生—临床医学]