Zhao Meng, Xu Qing, Yu Jiyun, Yu Yunzhou
Institute of Life Science and Biotechnology, Beijing Jiaotong University, Beijing 100044, China.
Sheng Wu Gong Cheng Xue Bao. 2011 Aug;27(8):1191-7.
Human immunodeficiency virus (HIV) infects the host cells by the fusion of viral and cell membranes. Blocking the combining between HIV and the receptors can prevent HIV from entering the host cells. We designed an invasion-inhibitor for HIV-1 targeting dendritic cell (DC), including 2 important HIV-1 receptors CD4 and CCR5, and 2 molecules Flt3-L and Mip-3alpha. With the synthetic gene of the inhibitor, 2 eukaryotic expression vectors pABK-CKR5-CD4/Flt3L-Mip3alpha (pABK-HIV-MF) and pABK-CKR5-CD4 (pABK-HIV-MT) were constructed and transfected into HEK 293 cells for expression. Results from RT-PCR, immunofluorescent assay, ELISA and Western blot approved that the invasion-inhibitor for HIV-1 was successfully and exactly expressed in the eukaryotic cells. Current study formed a solid base for the further research on the function of inhibitors for HIV-1 and elimination targeting DC.
人类免疫缺陷病毒（HIV）通过病毒膜与细胞膜融合来感染宿主细胞。阻断HIV与受体之间的结合可防止HIV进入宿主细胞。我们设计了一种针对树突状细胞（DC）的HIV-1入侵抑制剂，它包括两种重要的HIV-1受体CD4和CCR5，以及两种分子Flt3-L和Mip-3α。利用该抑制剂的合成基因，构建了两种真核表达载体pABK-CKR5-CD4/Flt3L-Mip3alpha（pABK-HIV-MF）和pABK-CKR5-CD4（pABK-HIV-MT），并将其转染到HEK 293细胞中进行表达。RT-PCR、免疫荧光分析、ELISA和蛋白质印迹法的结果证实，HIV-1入侵抑制剂在真核细胞中成功且准确地表达。当前的研究为进一步研究HIV-1抑制剂的功能以及针对DC的清除奠定了坚实基础。
