NEWLY RECOGNIZED LYMPHOCYTOTROPIC PARVOVIRUS OF MICE

新发现的小鼠嗜淋巴细胞细小病毒

• 批准号: 2285845
• 负责人: ABIGAIL L SMITH
• 金额: $ 27.66万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-03-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2285845
• 关键词: Parvoviridae; SCID mouse; antiviral antibody; athymic mouse; cellular immunity; cytotoxic T lymphocyte; delayed hypersensitivity; epitope mapping; epizootiology; flow cytometry; host organism interaction; humoral immunity; immunization; immunocytochemistry; in situ hybridization; laboratory mouse; lymphatic tissue; lymphocyte proliferation; mixed lymphocyte reaction test; polymerase chain reaction; transplant rejection; virulence; virus characteristic; virus protein; virus replication

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.

拟议研究的长期目标是表征体内 以及实验室新鉴定的体外嗜淋巴细胞细小病毒 小鼠，小鼠细小病毒（MPV）。 MPV 对生物医学构成重大威胁 使用小鼠进行研究，因为它会干扰免疫功能。然而， MPV 持续感染成年小鼠，因此它可以作为模型 持续性人类细小病毒感染。 具体目标是 (1) 继续MPV的分子表征，(2)扩展MPV的研究 发病机制，(3) 表征对 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) 鉴定可能影响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、南方和北方 印迹、免疫组织化学、原位杂交、病毒 定量、流式细胞术和评估体液和 细胞介导的免疫。抗体中和 MPV 的能力 被动给药时防止感染和/或持续存在 被确定。淋巴细胞在宿主反应中的作用是 在 SCID 和无胸腺小鼠以及免疫功能缺失的小鼠中进行评估 通过血清疗法产生 CD4+ 和/或 CD8+ T 细胞亚群。如果 T 细胞发挥 重要作用，将针对表位生成 MPV 特异性 T 细胞克隆 映射。将用选定的纯化蛋白质免疫小鼠 评估他们抵御活病毒挑战的能力。这 MPV 感染的功能影响将通过体外评估 增殖和细胞溶解试验以及小鼠体内 对蛋白质（卵清蛋白）抗原敏感，带有同种异体皮肤移植物的小鼠 或受到肿瘤攻击的小鼠。将检查的主机事实是 年龄（新生儿与断奶婴儿）和基因型，使用不同的遗传 近交系小鼠品系。