目的:建立一个基于细胞一细胞融合的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.