Rosenthal K S, Hodnichak C M, Summers J L
Cytometry. 1987 Jul;8(4):392-5. doi: 10.1002/cyto.990080408.
A rapid means of screening drugs for toxicity and anti-herpes simplex virus activity was developed based on the flow cytometric detection of HSV induced changes in cellular DNA content. Subconfluent monolayers of human diploid fibroblasts (HEL 299) were assayed for DNA content with propidium iodide 24 h after infection with HSV-1 (multiplicity of infection 1-10) and treatment with the drug to be tested. Infection was detected by a broadening of the normal diploid and tetraploid peaks and presence of greater than 4-n DNA staining. Inhibition of viral DNA synthesis and maintenance of the normal growth pattern of control cells was indication of antiviral activity. Toxicity of the compound was indicated by the loss of S phase and tetraploid cell populations. Using this assay, we evaluated the activities of one experimental and two established antiviral agents.
基于流式细胞术检测单纯疱疹病毒(HSV)诱导的细胞DNA含量变化,开发了一种快速筛选药物毒性和抗单纯疱疹病毒活性的方法。用人二倍体成纤维细胞(HEL 299)的亚汇合单层,在感染HSV-1(感染复数为1-10)并使用待测试药物处理24小时后,用碘化丙啶检测DNA含量。通过正常二倍体和四倍体峰变宽以及大于4倍体DNA染色的存在来检测感染。抑制病毒DNA合成并维持对照细胞的正常生长模式表明具有抗病毒活性。化合物的毒性通过S期和四倍体细胞群体的丧失来表明。使用该检测方法,我们评估了一种实验性抗病毒药物和两种已确立的抗病毒药物的活性。