Ciminale V, Felber B K, Campbell M, Pavlakis G N
National Cancer Institute-Frederick Cancer Research and Development Center, ABL-Basic Research Program, Frederick, MD 21702-1201.
AIDS Res Hum Retroviruses. 1990 Nov;6(11):1281-7. doi: 10.1089/aid.1990.6.1281.
The binding of human immunodeficiency virus type 1 (HIV-1) gp120env to CD4 is the first event leading to infection and represents an important target for possible therapeutic intervention. To provide a tool for screening and quantitation of the effects of drugs inhibiting the Env-CD4 interaction, we developed a simple, fast and quantitative bioassay measuring the fusion between two cell lines generated by stable transfection: one expressing high levels of HIV-1 proteins but no infectious virus (HL2/3), and the other expressing the CD4 receptor and containing an inducible chloramphenicol acetyltransferase (CAT) gene linked to the HIV-1 long terminal repeat (HLCD4-CAT). Upon cocultivation of HL2/3 and HLCD4-CAT cells, efficient cell fusion is observed within 8 h. The efficiency of fusion can be evaluated visually and quantitated by measuring CAT enzyme. This novel bioassay allows testing for drugs capable of interfering with the CD4-Env interaction. HL2/3 cell line secretes gp120env in the medium and can be used for the production of Env protein.
人类免疫缺陷病毒1型（HIV-1）包膜糖蛋白120（gp120env）与CD4的结合是导致感染的首个事件，也是可能的治疗干预的重要靶点。为了提供一种用于筛选和定量抑制Env-CD4相互作用药物效果的工具，我们开发了一种简单、快速且定量的生物测定法，该方法用于测量由稳定转染产生的两种细胞系之间的融合：一种表达高水平的HIV-1蛋白但无感染性病毒（HL2/3），另一种表达CD4受体并含有与HIV-1长末端重复序列相连的可诱导氯霉素乙酰转移酶（CAT）基因（HLCD4-CAT）。将HL2/3和HLCD4-CAT细胞共培养后，8小时内可观察到高效的细胞融合。融合效率可通过肉眼评估，并通过测量CAT酶进行定量。这种新型生物测定法可用于测试能够干扰CD4-Env相互作用的药物。HL2/3细胞系在培养基中分泌gp120env，可用于Env蛋白的生产。
