IN VITRO ANTIVIRAL SCREEN FOR HERPESVIRUSES

本合同的总体目标是提供资源， 新化学物质对成员的临床前体外评价 单纯疱疹病毒1型（HSV-1），疱疹病毒 单纯病毒2型（HSV-2）、巨细胞病毒（CMV）、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抑制试验筛选， HFF细胞对抗标准实验室适应菌株（EB病毒除外）。 将在HFF细胞中通过两种细胞评估初始细胞毒性。 增殖研究和中性红摄取测定。 EB病毒初筛 将通过免疫荧光法测量VCA或EA的产生 在用P3 HR-1 EBV超感染的Daudi或Raji细胞中的测定。 选择性指数为10或更高的物质将被 使用其他测定系统进行确认。 抗病毒活性（除 抗EBV的活性）将通过噬斑抑制试验证实， HFF细胞和来源于非人来源的其他细胞中。 抗EBV 活性将通过使用DNA杂交测定来确认， 抑制P3 HR-1细胞的DNA合成。 该电池 验证性测试旨在选择具有潜在 用于动物模型和临床试验的进一步评估。 选择进行额外后续研究的物质将 在许多不同的系统中进行评估，包括：（1）敏感性 对于另外的实验室传代病毒株和临床分离株， 包括耐药突变体;（2）联合化疗与其他 抗病毒药物;（3）作用机制研究;和（3）评价 使用骨髓克隆形成测定的毒性。

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.