一种拮抗HIV-1并靶向DC的融合基因的设计及表达鉴定

Design and expression of an inhibitor for HIV-1 targeting dendritic cell

摘要:

艾滋病病毒 (Human immunodeficiency virus，HIV) 通过与靶细胞膜的融合感染宿主细胞，研究表明阻断HIV与受体靶分子的结合可以阻止HIV进入宿主细胞，抑制HIV病毒的感染。设计合成了一个包含CD4和CCR5与HIV-1结合的主要功能结构区，及Flt3-L和Mip-3α分子的融合基因，构建了2个融合基因的真核表达载体pABK-CKR5-CD4/Flt3L-Mip3α (pABK-HIV-MF) 和pABK-CKR5-CD4 (pABK-HIV-MT)，在人胚肾293细胞中进行了表达。RT-PCR、细胞免疫荧光技术、ELISA和Western blotting检测结果表明融合基因在真核细胞中获得了正确的表达，这为进一步研究其对于HIV-1的拮抗并靶向树突状细胞 (DC) 清除研究奠定了基础。

Abstract:

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-3α. With the synthetic gene of the inhibitor, 2 eukaryotic expression vectors pABK-CKR5-CD4/Flt3L-Mip3α (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.