HIV-1假病毒感染未成熟树突状细胞实验研究

发布时间：2018-04-07 00:01

本文选题：HIV-假病毒　切入点：未成熟树突状细胞　出处：《中国免疫学杂志》2017年10期

【摘要】：目的:构建携带增强型绿色荧光蛋白基因(Enhanced green fluorescent protein,EGFP)的HIV-1假病毒;通过HIV-1假病毒感染未成熟树突状细胞(immatural dendritic cell,iDC)实验探讨病毒与细胞的相互作用。方法:利用慢病毒包装系统构建携带EGFP基因的HIV-1假病毒,通过RT-PCR扩增HIV-1假病毒中的EGFP基因;HIV-1假病毒感染iDC,对HIV-1假病毒感染iDC后的基因组整合和转录水平进行检测,观察被感染iDC中EGFP基因的表达。结果:构建的HIV-1假病毒通过RT-PCR结果显示扩增得到EGFP基因;HIV-1假病毒感染iDC后,在基因组整合和转录水平检测中都扩增得到了EGFP基因,荧光显微镜观察被HIV-1假病毒感染的iDC,观察到iDC表达绿色荧光蛋白。结论:成功构建携带EGFP基因的HIV-1假病毒;HIV-1假病毒可以感染iDC,并将其遗传物质整合到iDC基因组中,并经过转录和翻译使iDC表达EGFP。
[Abstract]:Aim: to construct HIV-1 pseudovirus carrying enhanced green fluorescent protein (EGFP) and to investigate the interaction between HIV-1 pseudovirus and immature dendritic cells infected with HIV-1 pseudovirus.Methods: HIV-1 pseudoviruses carrying EGFP gene were constructed by using lentivirus packaging system. The EGFP gene of HIV-1 pseudovirus was amplified by RT-PCR and infected with HIV-1 pseudovirus. The genomic integration and transcription level of HIV-1 pseudovirus infected with iDC were detected.To observe the expression of EGFP gene in infected iDC.Results: the constructed HIV-1 pseudovirus was amplified by RT-PCR, and the EGFP gene was amplified by the amplification of EGFP gene and HIV-1 pseudovirus infected with iDC in both genomic integration and transcriptional level detection.The expression of green fluorescent protein (GFP) was observed by fluorescence microscope in iDC infected by HIV-1 pseudovirus.Conclusion: HIV-1 pseudovirus carrying EGFP gene can infect iDCand integrate its genetic material into iDC genome, and iDC can express EGFP by transcription and translation.
【作者单位】：青岛市中心医院药学部;暨南大学第二临床医院深圳市人民医院;长春中医药大学基础医学院;军事医学科学院军事兽医研究所省部共建吉林省人兽共患病预防与控制重点实验室;
【基金】：国家自然科学基金(31472197) 病原微生物生物安全国家重点实验室开放课题(SKLPBS1435) 北京市自然科学基金(5152023) 吉林省青年科研基金项目(20140520173JH)资助
【分类号】：R512.91

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