TGF-β1基因修饰的树突状细胞移植对EAMG大鼠T淋巴细胞IFN-γ/IFN-γR表达的影响  

TGF-β1 Gene-modified Dendritic Cells Transplantation with Effectson the Expressions of IFN-γ and Its Receptor of T Lymphocytic Cellsin the Rats with Experimental Autoimmune Myasthenia Gravis

作　　者：王云甫[1] 孙圣刚[2] 李罗清 乔娴[2] 何国厚[1] 

机构地区：[1]郧阳医学院附属太和医院神经内科,湖北十堰442000 [2]华中科技大学同济医学院附属协和医院神经内科,湖北武汉430022 [3]岳阳市第一人民医院神经内科,湖南岳阳414000

出　　处：《中国神经免疫学和神经病学杂志》2009年第3期166-170,共5页Chinese Journal of Neuroimmunology and Neurology

基　　金：湖北省教育厅科学研究基金资助项目(2004Z001);湖北省自然科学研究基金资助项目(2006BA344)

摘　　要：目的探讨TGF-β1基因修饰的树突状细胞（Dc）移植对实验性自身免疫性重症肌无力（EAMG）大鼠T淋巴细胞IFN-γ分泌及其表面IFN-γ受体（IFN—γR）表达的影响。方法近交系、8～10周龄、健康雌性Lewis大鼠25只，随机分为健康对照组、EAMG组、DC对照组、pcDNA3-TGF-β1表达质粒转染的DC（pcDNA3-TGF-β1～DC）组与生理盐水对照组5组。除健康对照组外，各组均采用丁氏双鳍电鳐电器官乙酰胆碱受体（AChR）蛋白二次免疫方法复制EAMG大鼠模型。初次免疫后第5天分别皮下注射2×10^6DC、pcDNA3-TGF-β1—DC及等体积生理盐水，健康对照组和EAMG组不接受任何治疗。初次免疫后γ周，分离脾脏T淋巴细胞，分别应用酶联免疫吸附试验和免疫组化方法检测T淋巴细胞IFN-γ分泌及其表面IFN-γR表达水平。结果（1）体外培养48h后，健康对照大鼠T淋巴细胞上清液中IFN-γ水平很低，而EAMG组大鼠T淋巴细胞上清液中IFN-γ水平显著升高（P〈0．001）。pcDNA3-TGF-β1—DC治疗组IFN-γ水平与EAMG组差异无统计学意义（P〉0．05）。（2）健康对照大鼠T淋巴细胞经体外培养后可见少量IFN—γR阳性细胞，而EAMG组大鼠T淋巴细胞经体外培养后可见较多IFN—γR阳性细胞，其阳性细胞数及平均吸光度均明显高于健康对照组（P〈0．001）；pcDNA3-TGF—β1-DC组IFN-γR阳性细胞数较EAMG组明显增多，其阳性细胞平均吸光度亦明显高于EAMG组（均P〈0．01）。结论EAMG组大鼠T淋巴细胞IFN-γ分泌及其表面IFN-γR表达水平明显提高；pcDNA3-TGF-β1—DC治疗可进-步诱导IFN—γR表达，调节T淋巴细胞IFN—γ／IFN-γR表达可能是TGF-β1基因修饰的DC治疗EAMG的机制之-。Objective To study the effects of TGF-β1 gene-modified dendritic cells （DC） transplantation on the expressions of IFN-y and its receptor of T lymphocytic cells in the rats with experimental autoimmune myasthenia gravis （EAMG）. Methods Twenty-five healthy, female Lewis rats were divided randomly into 5 groups： normal control group, EAMG group, DCs control group, pcDNA3-TGF-β1 transfected dendritic cells （pcDNA3-TGFβ1- DCs） group and normal saline group. The rats were immunized with the protein extracted from electric organ of Narcine timilei and completed Freund＇s adjuvant （CFA） in the experiment groups except normal control group. On the 5th day after the initial immunization, 2 × 10^6 pcDNA3-TGFβ1-DCs/rat were injected subcutaneously into the backs of the rats in pcDNA3-TGFβ1-DCs treatment group, and the rats in DCs control group and normal saline group were injected in parallel with untreated DC and normal saline respectively. On the γth week after the initial immunization, T lymphocytic cells in every rat were separated from the spleen, the contents of IFN-γ/in the culture supernatants were detected by enzyme-linked immunoadsordent assay （ELISA）, and the expressions of IFN-γ receptor （IFN-TR） on the T lymphocytic cells were examined by using immunohistochemistry method. Results （1） Cultured for 48 h, the content of IFN-γ in the supernatant was extremely low found in the normal rats, and increased markedly in EAMG rats （P〈0. 001）. There was no significant difference in the contents of IFN-γ between the EAMG group and the pcDNA3-TGF-β1-DC group （P〉0.05）. （2） The IFN-γ/R positive cells were rare in normal rats, and increased significantly in EAMG rats （P〈0. 001）. The number and the average optical density of the IFN-TR positive cell in pcDNA3-TGF-β1-DC group were higher than that in EAMG group （P〈0. 01）, and it didn＇t show statistically difference among the EAMG group, DCs control group and normal saline group （P〉0. 05）. Conclusions

关 键 词：重症肌无力 转化生长因子-Β1 树突状细胞 细胞移植 Υ-干扰素 Υ-干扰素受体 

分 类 号：R746.1[医药卫生—神经病学与精神病学]