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表达CCR5Delta32基因对人外周血单核细胞增殖功能的影响 被引量：1

Effect of CCR5Delta32 Gene Expression in Human Peripheral Blood Mononuclear cells(PBMC) on Proliferation Function of the PBMC
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摘要 目的:在人外周血单核细胞(PBMC)内表达CCR5Delta32基因,研究是否对该细胞增殖功能产生影响。方法:构建pLenti-CCR5Delta32慢病毒载体,包装后产生重组慢病毒,将其转染PBMC,荧光显微镜观察转染情况,Westernblot鉴定目的蛋白的表达;分别用植物血凝素(PHA)及破伤风类毒素(TTD)刺激培养经转染的靶细胞,采用MTT比色法测定其增殖功能是否受影响。结果:构建了pLenti-CCR5Delta32慢病毒载体,经包装后产生重组慢病毒,滴度约为5×105TU/mL。将其转染PBMC,观察到约半数PBMC胞浆内有绿色荧光蛋白表达,经Westernblot进一步鉴定为目的蛋白;转染的靶细胞经PHA或TTD刺激培养后,有着与正常PBMC相似的增殖功能。结论:构建了重组慢病毒并将其转染PBMC靶细胞,为获得性免疫缺陷综合征的基因治疗研究奠定了基础。 Objective： To explore the effect of CCR5Delta32 gene expression in human peripheral blood mononuclear cells（PBMC） on proliferation function of the PBMC. Methods： The recombinant lentivirus containing human CCR5Delta32 gene was constructed using genetic engineering technology and then transfected into human PBMC. The expression of CCRSDelta32 gene was detected by Western blot and the PBMC proliferation function induced by plant haemagglutinin （PHA） and tetanus toxoid（TTD） was determined with methyl thiazolyl tetrazolium（MTT） assay. Results： The recombinant lentivirus containing human CCRSDelta32 gene was constructed successfully, and its titer reached 5×10^5 TU/mL. After transfected into PBMC, the target gene was showed expression. Proliferation function of the PBMC transfected with the recombinant lentivirus was similar with normal PBMC. Conclusion： Our present work lays a foundation of further studying the gene therapy for acquired immunodeficiency syndrome.
作者 安群星 穆士杰 张献清 陈蕤 易静 余瑞
机构地区 第四军医大学西京医院输血科
出处 《生物技术通讯》 CAS 2009年第3期323-325,共3页 Letters in Biotechnology
基金 国家自然科学基金(30571675)
关键词 CCR5Delta32基因 慢病毒载体 外周血单核细胞 人类免疫缺陷病毒 辅受体 CCRSDelta32 gene lentiviral vector peripheral blood mononuclear cell human immunodeficiency virus coreceptor
分类号 Q78 [生物学—分子生物学]
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生物技术通讯
2009年 第3期

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