A NEWLY RECOGNIZED LYMPHOCYTOTROPIC PARVOVIRUS OF MICE

这项拟议的研究的长期目标是在活体内表征 和体外新发现的实验室嗜淋巴细胞细小病毒 小鼠，小鼠细小病毒(MPV)。MPV是对生物医学的重大威胁 使用老鼠进行研究，因为它干扰免疫功能。然而， MPV持续感染成年小鼠，因此可作为一种模型 持续的人类细小病毒感染。具体目标是(1) 继续MPV的分子表征，(2)扩大MPV的研究 发病机制，(3)表征MPV的免疫反应，(4) 扩大与MPV感染相关的功能影响的研究，以及 (5)确定可能影响MPV表达的宿主因素 感染。使用的方法包括细菌表达载体 系统，聚合酶链式反应，免疫印迹，南方和北方 印迹、免疫组织化学、原位杂交、病毒 定量、流式细胞术和分析以评估体液和 细胞免疫。抗体中和MPV和MPV的能力 在被动给药时防止感染和/或持久性 要下定决心。淋巴样细胞在宿主反应中的作用将是 在SCID和无瘤小鼠以及免疫能力耗竭的小鼠中进行评估 通过血清疗法检测CD4+和/或CD8+T细胞亚群。如果T细胞发挥作用 重要作用，将产生MPV特异性T细胞克隆用于表位 映射。将用精选的纯化蛋白免疫小鼠 评估他们抵御活病毒挑战的能力。这个 MPV感染的功能效应将在体外用 对符合以下条件的小鼠进行增殖和细胞溶解试验 对蛋白质(卵蛋白)抗原致敏的小鼠，移植皮肤 或者是受到肿瘤挑战的小鼠。将审查的主机事实包括 年龄(新生儿与断奶)和基因，使用不同的基因 近交系小鼠品系。

The long-term goal of the proposed research is to characterize in vivo and in vitro a newly recognized lymphocytotropic parvovirus of laboratory mice, mouse parvovirus (MPV). MPV is a significant threat to biomedical research using mice because it interferes with immune function. However, MPV persistently infects adult mice, so it may serve as a model for persistent human parvovirus infection. The specific aims are (1) to continue molecular characterization of MPV, (2)to extend studies of MPV pathogenesis, (3) to characterize the immune response to MPV, (4) to extend studies of functional effects associated with MPV infection, and (5) to identify host factors that may influence expression of MPV infection. Methods to be used include bacterial expression vector systems, polymerase chain reaction, immunoblotting, Southern and Northern blotting, immunohistochemistry, in situ hybridization, virus quantification, flow cytometry, and assays to assess humoral and cell-mediated immunity. The capacity of antibody to neutralize MPV and to prevent infection and/or persistence when passively administered will be determined. The role of lymphoid cells in the host response will be evaluated in SCID and athymic mice and in immunocompetent mice depleted of CD4+ and/or CD8+ T cell subsets by serotherapy. If T cells play an important role, MPV-specific T cell clones will be generated for epitope mapping. Mice will be immunized with selected purified proteins to evaluate their capacity to protect upon live virus challenge. The functional effects of MPV infection will be assessed in vitro using proliferative and cytolysis assays and in vivo among mice that are sensitized to protein (ovalbumin) antigen, mice that bear skin allografts or mice that are tumor-challenged. Host facts that will be examined are age (neonate vs. weanling) and genotype, using genetically disparate inbred mouse strains.