摘要
目的:建立一个基于细胞-细胞融合的HIV进入抑制剂非感染性筛选方法。方法:将稳定表达HIV包膜蛋白gp160的效应细胞与含有或稳定表达HIV受体和辅助受体的靶细胞混合,观察合胞体的形成。用特异性的HIV进入抑制剂对检测方法进行验证。结果:本研究比较了2种效应细胞与5种靶细胞的10种组合,发现将CHO-WT和MT-2细胞混合,可形成明显的合胞体。进一步发现特异性的HIV进入抑制剂能够抑制合胞体的形成,且具有剂量依赖性。结论:这一方法能特异性地筛选作用在HIV进入阶段的抗HIV药物,操作简单方便,且无感染性,可望发展成为一个无感染性的多靶点高内涵药物筛选模型,用于天然和合成来源的小分子HIV进入抑制剂的筛选。
Aim： To develop a non-infectious cell-cell fusion assay that can be used in a regular biological laboratory for high throughput screening for HIV entry inhibitors. Methods： Cells expressing HIV envelope glycoprotein gpl60 （ as effector ceils） and expressing receptor and co-receptor for HIV （as target cells）, respectively, were mixed and cultured before the syncytium formation was recorded. Specific HIV entry inhibitors were used to validate the established detecting method. Results： We compared the syncytium formation activities of 10 combinations of 2 effector cell lines and 5 target cell lines, respectively, and found that mixing of CHO-WT and MT-2 cells could form syncytia obviously. Further studies showed that HIV entry inhibitors could inhibit the syncytium formation in a dose-dependent manner. Conclusion：We have developed a simple and non-infectious cell-cell fusion assay that can be used to screen for novel HIV entry inhibitors from natural and synthetic compound libraries.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2007年第6期576-580,共5页
Journal of Jinan University(Natural Science & Medicine Edition)
基金
国家自然科学基金项目(30672496)
广东省科技攻关项目(2005A10904002)
广州市科技攻关项目(2005Z2-E4041)
