PATHOGENESIS OF ALEUTIAN MINK DISEASE PARVOVIRUS INFECTIONS

阿留申水貂病细小病毒持续感染水貂 (ADV)导致免疫调节紊乱，包括 高丙种球蛋白血症，浆细胞增多症，免疫复合体疾病， 间质性和肾小球肾炎。主要目标细胞群 用于病毒复制和隔离的是巨噬细胞。感染是 被抗病毒抗体增强，但在某种程度上被限制在 单个细胞的。这个项目的目的是阐明 病毒感染引起这种疾病的致病机制。 今年的研究总结包括： *对ADV非结构蛋白的抗体反应。感染ADV的水貂 针对病毒常见的氨基酸序列产生抗体 非结构蛋白，NS1、NS2和NS3。尽管针对该病毒的抗体 NS1的独特区域也存在，没有针对独特的抗体 检测到NS2或NS3区。 *ADV蛋白在允许感染中的亚细胞定位。在……里面 在感染后期核溶解之前被允许感染的CRFK细胞， AdV病毒粒子蛋白(VP)形成壳状核内结构 病毒DNA、NS1和NS2的周围包涵体。NS2(约50%)也是 在细胞质中发现的，表现出一些穿梭的特征 像HIV rev蛋白一样的蛋白质。 *ADV对单核细胞系K562的感染有抑制作用。副总裁、NS1和NS2 在抗体依赖感染K562细胞的细胞核中发现 使用ADV。与CRFK的允许ADV感染不同，NS2在 K562的细胞质。ADV感染使正常细胞周期停滞 K562在晚期，类似于CRFK中的细胞周期中断。 在ADV中未观察到IL-6蛋白的上调 感染K562细胞。 *水貂外周血单核细胞(PBMC)的ADV感染。一个 建立了水貂外周血单核细胞感染ADV的研究体系。跟随 ADV感染未感染水貂的纯化PBMC，几乎全部 单核细胞显示特异的胞浆荧光。很低的百分比 在后来的时间点上继续展示了抗原的核定位， 表明病毒复制有限。 *聚合酶链式反应(PCR)与对抗免疫的比较 电泳法(CEP)在ADV感染诊断中的应用CEP被发现 在商业广告中检测ADV感染比PCR更敏感 水貂牧场布景。 *ADV感染水貂的细胞因子反应。测定细胞因子水平 用非放射性核糖核酸酶对ADV感染水貂淋巴组织的研究 保护试验(RPA)。IL-6没有观察到一致的模式， 感染与未感染患者IL-10、IL-1、IL-12 p35、IL-12 p40、肿瘤坏死因子的比较 动物。

Persistent infection of mink with the Aleutian mink disease parvovirus (ADV) leads to disturbances of immune regulation, including hypergammaglobulinemia, plasmacytosis, immune complex disease, interstitial and glomerulonephritis. The major target cell population for viral replication and sequestration is macrophages. Infection is enhanced by anti-viral antibody, but is somehow restricted at the level of the individual cell. The purpose of this project is to elucidate the pathogenetic mechanisms by which viral infection causes this disorder. A summary of this year's studies includes: * Antibody response to ADV nonstructural proteins. ADV infected mink made antibody against amino acid sequences common to the viral nonstructural proteins, NS1, NS2 and NS3. Although antibody to the unique regions of NS1 was also present, no antibody against the unique regions of NS2 or NS3 was detected. * Subcellular localization of ADV proteins in permissive infection. In permissively infected CRFK cells prior to nucleolysis late in infection, ADV virion proteins (VP) formed shell-like intranuclear structures surrounding inclusions of viral DNA, NS1 and NS2. NS2 (ca. 50%) was also found in the cytoplasm, exhibiting some characteristics of a shuttling protein like HIV rev protein. * ADV restricted infection of monocyte cell line, K562. VP, NS1 and NS2 were found in nuclei of K562 cells after antibody dependent infection with ADV. Unlike permissive ADV infection of CRFK, NS2 was not noted in the cytoplasm of K562. ADV infection arrested the normal cell cycle of K562 in later stages, similar to cell cycle disruption seen in CRFK. Upregulation of interleukin 6 protein (IL-6) was not observed in ADV infected K562 cells. * ADV infection of mink peripheral blood mononuclear cells (PBMC). A system for studying ADV infection in mink PBMC was developed. Following ADV infection of purified PBMC from uninfected mink, virtually all monocytes exhibited specific cytoplasmic fluorescence. A low percentage went on to exhibit nuclear localization of antigen at later time points, indicating limited viral replication. * Comparison of polymerase chain reaction (PCR) with counterimmune electrophoresis (CEP) in diagnosis of ADV infections. CEP was found to be more sensitive than PCR for detecting ADV infections in a commercial mink ranch setting. * Cytokine response of ADV infected mink. Cytokine levels were assayed on lymphoid tissues from ADV infected mink using a non-radioactive RNAse protection assay (RPA). No consistent pattern was observed for IL-6, IL-10, IL-1, IL-12 p35, IL-12 p40 or TNF between infected and uninfected animals.