树突状细胞负载Tα146-162预防实验性自身免疫性重症肌无力的实验研究  

Prevention of experimental autoimmune myasthenia gravis by dendritic cell pulsed with Tα146-162

作　　者：胡珏[1] 杨欢[1] 肖波[1] 黄颉[1] 李水仙[1] 李静[1] 周文斌[1] 

机构地区：[1]中南大学湘雅医院神经内科,长沙410008

出　　处：《中华神经科杂志》2006年第4期224-228,共5页Chinese Journal of Neurology

基　　金：国家自然科学基金资助项目(30570639)

摘　　要：目的探讨负载电鳗乙酰胆碱受体(TAChR)免疫优势肽段α146-162(Tα146-162)的不成熟髓源树突状细胞(iMDC),即Tα146-162-iMDC对实验性自身免疫性重症肌无力(EAMG)的预防作用及其机制。方法(1)24只C57BL/6小鼠随机分为Tα146-162-iMDC组、iMDC组、对照组、正常组,每组6只。Tα146-162-iMDC组、iMDC组和对照组小鼠皮下注射TAChR抗原乳剂(作为第0天)。第7天,Tα146-162-iMDC组皮下注射Tα146-162-iMDC,iMDC组和对照组分别注射对照iMDC和磷酸盐缓冲液(PBS)。正常组小鼠不做干预。第14天检测淋巴细胞增殖反应。(2)另外24只C57BL/6小鼠随机分为预防组和EAMG组。每只小鼠在第0、30、60天皮下注射抗原乳剂诱导EAMG。预防组每次免疫后3d皮下注射Tα146-162-iMDC,对照组皮下注射PBS。观察发病情况并进行肌力评分。第90天结束观察。(3)3H-TdR掺入法检测淋巴细胞增殖反应,酶联免疫吸附实验检测淋巴细胞培养液上清中干扰素(IFN)-γ、白细胞介素(IL)-6的含量。结果(1)Tα146-162-iMDC组、iMDC组、对照组和正常组TAChR特异性淋巴细胞增殖反应值(放射性荧光闪烁计数)分别为3314·1±571·3、7717·5±1283·4、8608·6±1458·9、922·9±109·3,与iMDC组和对照组相比,Tα146-162-iMDC组抗原特异性淋巴细胞增殖反应显著降低(P<0·01)。(2)预防组和EAMG组临床评分分别为(0·33±0·65)、(1·46±1·08)分,差异有统计学意义(P<0·01);预防组和EAMG组发病率分别为3/12(25%)和9/12(75%)只,差异有统计学意义(P<0·05)。(3)预防组和EAMG组TAChR特异性淋巴细胞增殖反应值分别为3347·6±556·9、9416·2±1546·5,预防组显著低于EAMG组(P<0·01)。(4)预防组和EAMG组淋巴细胞在TAChR刺激下分泌IFN-γ分别为(117·6±18·5)、(363·1±63·2)pg/ml,分泌IL-6为(49·7±9·9)、(90·6±13·6)pg/ml;与EAMG组相比,预防组淋巴细胞抗原特异性IFN-γ、IL-6分泌显著减少(P<0·01)。结论Tα146-162-iMDC能抑制TAChR预致敏T细胞免疫�Objective To explore the preventing potential and mechanism of immature myeloid dendritic cell （iMDC） pulsed with immunodominant peptide α146-162 of Torpedo acetylcholine receptor （TAChR, Tα146-162-iMDC） in experimental autoimmune myasthenia gravis （EAMG）. Methods （1） C57BL/6 mice were randomly enrolled into Tα146-162-iMDC group, iMDC group, control group and normal group respectively （n = 6）. On day 0, Tα146-162-iMDC group, iMDC group, control group were immunized with TAChR in CFA. On day 7, the mice of Tα146-162-iMDC group were subcutaneously injected with Tα146-162-iMDC. iMDC group and control group were subcutaneously injected with iMDC and PBS respectively. Normal group received no treatment. On day 14, lymphocyte proliferation of the four groups was measured. （2） C57BL/6 mice were randomly enrolled into prevention group and EAMG group （n=12）. On day 0, 30, 60, mice were immunized with TAChR in CFA to inducing EAMG. Mice in prevention group were subcutaneously injected with Tα146-162-iMDC 3 days later after each immunization while EAMG group received subcutaneous injection of PBS only. Clinical score of myasthenia were evaluated till the 90th days. （3） After the termination of experiment, lymphocyte proliferation were assayed. Levels of IFN-γ and IL-6 in the supernatants of leukomonocyte culture were measured by enzyme-linked immunosorbent assay. Results （ 1 ） The lymphocyte proliferative response after TAChR stimulation of Tα146-162-iMDC group, iMDC group, control group and normal group was （3314. 1 ±571.3）, （7717. 5 ± 1283.4） , （8608.6 ± 1458. 9） , （922.9 ± 109. 3）cpm respectively. Compared with iMDC group and control group, Tα146-162-iMDC group had a significantly suppressed antigen specific lymphocyte proliferation （ P 〈 0. 01 ）. （2） The average clinical score of prevention group and EAMG group was 0. 33 ±0. 65 and 1.46 ± 1.08 respectively. The accumulated incidence in prevention group and EAMG group was 25% and 75% respective

关 键 词：树突细胞 受体 胆碱能 免疫耐受 重症肌无力 实验性自身免疫性 

分 类 号：R746.1[医药卫生—神经病学与精神病学] R392.12[医药卫生—临床医学]