CCR5~Δ32及CCR5 siRNA共修饰的树突状细胞抗HIV-1的初步研究

The preliminary study of CCR5~Δ32 and CCR5 siRNA modified dendritic cells resistant HIV-1 infection

目的：分析CCR5^△32及CCR5 siRNA共修饰的树突状细胞抗人类免疫缺陷病毒Ⅰ型（HIV-1）的作用。方法采用Adeasy系统构建携带CCR5^△32及CCR5 siRNA的重组腺病毒载体；在体外将人外周血单个核细胞（PBMC）发育成树突状细胞；采用免疫印迹法分析细胞内CCR5^△32、CCR5及HIV-1 p24的表达情况；采用酶联免疫吸附测定（ELISA）分析HIV-1 p24含量。结果重组腺病毒载体感染人源性PBMC后，细胞内CCR5表达下降，CCR5^△32表达增加。HIV-1感染PBMC后，经修饰后的细胞中p24含量较低，而未修饰的细胞中p24含量持续上升。HIV-1感染PBMC来源的树突状细胞后，经修饰后的细胞中p24含量较低，而未修饰的细胞中p24含量持续上升。结论 CCR5^△32及CCR5 siRNA共修饰的树突状细胞具有抗HIV-1的功能。但将修饰后的细胞回输入体内能否重建机体免疫系统功能以及载体的安全性和高效性如何评价，尚需要进一步研究。

Objective To identify the characteristics of recombinant adenovirus modiifed PBMC-derived dendritic cells to resist the HIV-1 infection. Method The recombinant adenovirus vector was integrated with CCR5^△32 and CCR5 siRNA genes by using Adeasy system. The human PBMC from healthy donor blood was isolated in order to get dendritic cells. The expression of CCR5^△32, CCR5 and HIV-1 p24 in PBMC or modified cells was measured by Western blot, and ELISA. Result After the cells had been modified by recombinant adenovirus vector, the expression of CCR5^△32 was increased while the expression of CCR5 was decreased. The expression of p24 was decreased when the cells had been modified by recombinant adenovirus vector compared to the un-modiifed cells. The modiifed cells showed resistance to HIV-1 infection. Conclusion The recombinant adenovirus-modiifed cells show good resistance to HIV-1 infection. The effect and safety of modiifed cells need to be explored by further researches.