Pauwels R, De Clercq E, Desmyter J, Balzarini J, Goubau P, Herdewijn P, Vanderhaeghe H, Vandeputte M
Rega Institute for Medical Research, Katholieke Universiteit Leuven, Belgium.
J Virol Methods. 1987 Jun;16(3):171-85. doi: 10.1016/0166-0934(87)90002-4.
Human immunodeficiency virus (HIV) infection of MT-4 cells, an HTLV-I-transformed T-cell line, proved to be a rapid and sensitive assay system for the detection of potential antiviral drugs effective against the acquired immune deficiency syndrome (AIDS). Four days after HIV inoculation of the MT-4 cells, viral antigen expression was monitored in parallel with indirect immunofluorescence microscopy and laser flow cytofluorography. When 3'-azido-2',3'-dideoxythymidine (AzddThd, AZT) and 2',3'-dideoxycytidine (ddCyd) were evaluated under these conditions, they inhibited viral antigen expression at a minimum (33% inhibitory) concentration of 0.0004 and 0.02 microM, respectively. Similar minimum effective concentrations were found for AzddThd and ddCyd in assays where inhibition of viral cytopathogenicity was based on cell survival. While laser flow cytofluorography could be best adapted for quantitative measurements, cell survival and reconstitution of disrupted cell aggregates gave an equally rapid and sensitive endpoint; and the latter may be ideally suited for preliminary drug screening.
MT-4细胞是一种由人嗜T淋巴细胞病毒I型(HTLV-I)转化的T细胞系,人免疫缺陷病毒(HIV)对其感染被证明是一种快速且灵敏的检测系统,可用于检测对获得性免疫缺陷综合征(AIDS)有效的潜在抗病毒药物。用HIV接种MT-4细胞4天后,采用间接免疫荧光显微镜和激光流式细胞荧光术同时监测病毒抗原表达。在这些条件下评估3'-叠氮-2',3'-双脱氧胸苷(AzddThd,AZT)和2',3'-双脱氧胞苷(ddCyd)时,它们分别在最低(33%抑制)浓度0.0004和0.02微摩尔时抑制病毒抗原表达。在基于细胞存活来检测病毒致细胞病变效应抑制情况的实验中,也发现了AzddThd和ddCyd类似的最低有效浓度。虽然激光流式细胞荧光术最适合进行定量测量,但细胞存活以及破碎细胞聚集体的重建给出了同样快速且灵敏的终点;而后者可能最适合用于初步药物筛选。