NEWLY RECOGNIZED LYMPHOCYTOTROPIC PARVOVIRUS OF MICE

拟议研究的长期目标是在体内表征 和体外实验室新发现的亲淋巴细胞细小病毒 小鼠，小鼠细小病毒（MPV）。MPV是生物医学的重大威胁 因为它会干扰免疫功能。然而，在这方面， MPV持续感染成年小鼠，因此它可以作为 持续性人类细小病毒感染。 具体目标是：（1） 继续进行MPV的分子鉴定，（2）扩大MPV的研究 发病机制，（3）表征对MPV的免疫应答，（4） 扩展与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，聚合酶链反应，免疫印迹，Southern和北方 印迹，免疫组织化学，原位杂交，病毒 定量，流式细胞术和测定，以评估体液和 细胞免疫抗体中和MPV的能力， 为了防止感染和/或当被动施用时的持久性， 被确定。淋巴细胞在宿主反应中的作用将是 在SCID和无胸腺小鼠以及免疫活性小鼠中进行了评价， CD 4+和/或CD 8 + 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.小鼠将用选择的纯化蛋白免疫， 评估其在活病毒攻击下的保护能力。的 将使用体外评估MPV感染的功能影响 增殖和细胞溶解测定以及在小鼠体内， 对蛋白质（卵清蛋白）抗原致敏的小鼠， 或者是肿瘤激发的小鼠。将检查的主机事实包括 年龄（新生儿vs.断奶）和基因型，使用遗传差异 近交系小鼠

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.