IN VITRO ANTIVIRAL SCREEN FOR HERPESVIRUSES

本合同的总体目标是为 新化学物质对成员的临床前体外评价 疱疹病毒组中--单纯疱疹病毒1型(HSV-1)，疱疹 单纯疱疹病毒2型、巨细胞病毒、水痘-带状疱疹 病毒(VZV)和EB病毒(EBV)。在最初的研究中， 实验化合物将使用CPE抑制试验进行筛选 HFF细胞对实验室适应的标准毒株(EBV除外)的作用。 两种细胞都将评估HFF细胞的初始细胞毒性 细胞增殖实验和中性红摄取实验。最初的EBV筛查 将通过免疫荧光法测量VCA或EA的产生 在重叠感染P3HR-1 EBV的Daudi或Raji细胞中检测。 选择性指数为10或更大的物质将被 已使用其他化验系统确认。抗病毒活性(除 对EBV的活性)将通过斑块抑制试验在#年确认 HFF细胞和来自非人类来源的其他细胞。抗EB病毒 活性将通过使用DNA杂交试验来确认 抑制P3HR-L细胞的DNA合成。这块电池 验证性试验的目的是选择有潜力的物质 在动物模型和临床试验中进行进一步评估。 被选中进行额外后续研究的物质将是 在许多不同的系统中进行评估，包括：(1)敏感性 更多实验室传代的病毒株和临床分离株， 包括耐药突变株；(2)与其他 抗病毒药物；(3)作用机制研究；以及(3)评价 使用骨髓克隆形成试验进行毒性试验。

The overall objective of this contract is to provide a resource for preclinical in vitro evaluation of new chemical substances against members of the herpesvirus group -- herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), cytomegalovirus (CMV), varicella-zoster virus (VZV), and Epstein-Barr virus (EBV). In the initial studies, experimental compounds will be screened using the CPE inhibition assay in HFF cells against standard laboratory-adapted strains (except EBV). Initial cytotoxicity will be assessed in HFF cells by both cell proliferation study and neutral red uptake assay. The initial EBV screen will be to measure the VCA or EA production by the immunofluorescence assay in Daudi or Raji cells superinfected with P3HR-1 EBV. Substances that have a selectivity index of ten or greater will be confirmed using additional assay systems. Antiviral activity (except activity against EBV) will be confirmed by the plaque inhibition assay in HFF cells and in other cells derived from non-human sources. The anti-EBV activity will be confirmed by using the DNA hybridization assay to measure the inhibition of DNA synthesis in P3HR-l cells. This battery of confirmatory tests is designed to select substances that have potential for further evaluation in animal models and clinical trials. Substances that are selected for additional follow-up studies will be evaluated in a number of different systems, including: (1) susceptibility to additional laboratory passaged viral strains and clinical isolates, including drug-resistant mutants; (2) combination chemotherapy with other antiviral agents; (3) mechanism of action studies; and (3) evaluation of toxicity using a bone marrow clonogenic assay.