In Vitro Study on Vertical Transmission of the HIV-1 gag Gene by Human Sperm
In Vitro Study on Vertical Transmission of the HIV-1 gag Gene by Human Sperm
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HIV-1 gag 基因通过人类精子垂直传播的体外研究
DOI：
10.1002/jmv.21767
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发表时间：
2011-01-01
影响因子：
12.7
通讯作者：
Huang, Tian-Hua
中科院分区：
文献类型：
--
作者：
Wang, Dian;Kang, Xiang-Jin;Huang, Tian-Hua
艾滋病毒/艾滋病是世界范围内的一个主要公共卫生问题。为探讨人类精子垂直传播HIV的可行性，进行了质粒构建和转基因、人精子体外受精、荧光原位杂交(FISH)、逆转录-聚合酶链式反应(RT-PCR)和免疫荧光试验(IFA)。在转基因人精子的核和染色体、受精卵的男性原核和双细胞胚胎的卵裂球核中观察到HIV-1 Gag DNA的FISH信号，表明HIV-1 Gag基因可以通过精子膜传递并整合到精子基因组中。相比之下，携带靶基因的人类精子实现了正常受精，精子介导的靶基因的复制与宿主基因组同步。RT-PCR检测到目的基因在转基因人精子和两细胞胚胎中均有阳性条带。这些结果进一步证实了靶基因可以在人类精子和胚胎细胞中转录成mRNA。IFA结果显示，在含有精子介导靶基因的双细胞胚胎中，HIV-1 p24 Gag蛋白呈阳性信号，而在转基因的人精子中不表达，这表明精子介导靶基因可以在胚胎细胞中翻译成HIV-1 p24 Gag蛋白，但在精子细胞中不能。这一结果为HIV-1 Gag基因可能通过体外受精而垂直传播到胚胎提供了证据。J.Med.维罗尔。83：16-23,2010。(C)2010年Wiley-Liss公司
HIV/AIDS is a major public health problem worldwide. To explore the feasibility of HIV vertical transmission by human sperm, plasmid construction and transfection, interspecific in vitro fertilization of zona-free hamster ova by human sperm, fluorescence in situ hybridization (FISH), RT-PCR, and immunofluorescence assay (IFA) were carried out. The FISH signals for HIV-1 gag DNA were observed in the nuclei and chromosomes of transfected human sperm, male pronuclei of zygotes, and nuclei of blastomeres of two-cell embryos, indicating that the HIV-1 gag gene could be transmitted via the sperm membrane and integrated into the sperm genome. In contrast, human sperm carrying the target gene achieved normal fertilization, and replication of the sperm-mediated target gene was synchronized with the host genome. Using RT-PCR, the positive bands for the target gene were observed in the transfected human sperm and two-cell embryos. These results further confirm that the target gene can be transcribed into mRNA in human sperm and embryonic cells. Positive signals for the HIV-1 p24 gag protein were shown by IFA in two-cell embryos containing the sperm-mediated target gene and not in the transfected human sperm, which indicated that the sperm-mediated target gene could be translated to make HIV-1 p24 gag protein in embryonic cells, but not in sperm cells. The results provide evidence for possible vertical transmission of the HIV-1 gag gene to the embryo by fertilizing sperm in vitro. J. Med. Virol. 83:16-23, 2010. (C) 2010 Wiley-Liss, Inc.