重组腺病毒修饰外周血单核细胞来源的树突状细胞对人类免疫缺陷病毒-1抗感染作用研究  

Recombinant Adenovirus Modified PBMC-Derived Dendritic Cells are Resistant to HIV-1 Infection

作　　者：夏承来[1] 罗红彬[2] 黄汉辉[1] 梅峥嵘[1] 严鹏科[1] 

机构地区：[1]广州医学院第三附属医院药学部,广州510150 [2]广州市第八人民医院,广州510006

出　　处：《中国药学杂志》2012年第12期960-964,共5页Chinese Pharmaceutical Journal

基　　金：广东省计生委项目(20110266;20110316)

摘　　要：目的利用腺病毒载体技术,研究目的基因修饰的人源性外周血单核细胞来源的树突状细胞对抗人类免疫缺陷病毒-1的感染。方法将目的基因CCR5Δ32,CCR5siRNA,人类免疫缺陷病毒-1 poli和人类免疫缺陷病毒-1 inti整合到腺病毒载体上,在体外将人源性外周血单核细胞诱导为树突状细胞。用免疫印迹检查目的基因CCR5、CCR5Δ32和CXCR4的表达情况。并通过检测人类免疫缺陷病毒-1 p24抗原的表达来判断修饰后的细胞对人类免疫缺陷病毒-1病毒的抵抗情况。结果用重组腺病毒修饰后的细胞中,CCR5Δ32的表达明显增加,CCR5和CXCR4的表达明显降低。当修饰后的细胞发育成树突状细胞后,p24的表达水平低于未修饰的细胞,说明细胞被修饰后对人类免疫缺陷病毒-1的抵抗能力明显增加。结论利用重组腺病毒修饰的细胞能够较好的抵抗人类免疫缺陷病毒-1的感染。对人源性干细胞来源的免疫细胞如树突状细胞进行修饰,可望成为治愈人类免疫缺陷病毒-1的潜在手段。OBJECTIVE To identify the characteristics of recombinant adenovirus modified PBMC-derived dendritic ceils to resist the HIV-1 infection. METHODS The recombinant adenovirns vector was integrated with CCRSA32, CCRSsiRNA, HIV-1 poli and HIV-1 inti genes by using AdEasy system. The human PBMC from healthy donor blood was isolated in order to get dendritic ceils. The expression of CCRSA32, CCRS, CXCR4 and HIV-1 p24 in PBMC or modified cells was measured by Westernblot. RESULTS After the cells have been modified by Ad-RSA32siHIV, the expression of CCRSA32 was increased while the expression of CCR5 and CXCR4 were decreased. The level of p2A was lower in modified cells than that of unmodified cells. The modified ceils show resistance to HIV- 1 infection. CONCLUSION The recombinant adenovirus-modified cells show good resistance to HIV-1 infection. Modification of HSC-derived immunity ceils, such as DCs. mav be a potent strategy to resist HIV-1 infection.

关 键 词：重组腺病毒 人类免疫缺陷病毒-1 树突状细胞 基因治疗 外周血单核细胞 

分 类 号：R965[医药卫生—药理学]