siRNA沉默Rac2蛋白表达下调树突状细胞的交叉递呈  

Rac2 influences the cross-presentation of dendritic cells

作　　者：李娜[1] 邹丽云[1] 万瑛[1] 张晋宇[1] 刘婷[1] 李景怡[1] 吴玉章[1] 

机构地区：[1]第三军医大学基础部全军免疫学研究所,重庆400038

出　　处：《免疫学杂志》2008年第3期272-274,278,共4页Immunological Journal

摘　　要：目的通过RNA干扰研究Rac2蛋白对树突状细胞(DC)交叉递呈的影响。方法首先构建针对Rac2基因siRNA的慢病毒载体pFIVsiRNARac2-1,pFIVsiRNARac2-2,pFIVsiRNARac2-3;DNA-磷酸钙共沉淀的方法转染293FT细胞包装慢病毒,嘌呤霉素(puromycin)筛选病毒感染的阳性DC2.4(树突状细胞系),Western-blot检测其干扰效率;然后用B3Z细胞(H2-Kb限制性,SIINFEKL特异性T细胞杂交瘤)检测筛选后的DC2.4细胞交叉递呈的能力。结果酶切和测序结果证实表达siRNA的慢病毒载体构建成功,抗原递呈实验发现下调了Rac2蛋白表达的DC2.4细胞交叉递呈能力下降。结论初步证实了Rac2蛋白参与树突状细胞对外来抗原的交叉递呈,为进一步了解树突状细胞交叉递呈的分子机制奠定了基础。Objective To investigate the role of Rac2 in cross-presentation of dendritic cells by RNA interference technology. Methods ①Lentivirus vectors for expression of Rac2 siRNA were constructed. ②Lentivims particles were produced by co-transfecfion with second-generation packaging plasmid in 293FT. ③ The interference efficiency of Rac2 expression was detected by Western-blotting. ④ After transfection, puromycin selection, and phagocytosis of exogenous antigen, the ability of cross-presentation in DC2.4 cell line was measured. Results The construction of lentivirus vectors for expression of Rac2 siRNA was confmned by restriction enzyme and DNA sequencing. The puromycin-selected DC2.4 cell line, after being transfected by lentivirus, dramatically decreased the IL-2 secretion in cross-presentation assay. The primary experiments indicate that Rac2 is involved in cross-presentation of dendritic cells, which provides a foundation for understanding the molecular mechanisms of cross-presentation.

关 键 词：树突状细胞 Rat2 交叉递呈 

分 类 号：R392[医药卫生—免疫学]