PATHOGENESIS OF ALEUTIAN MINK DISEASE PARVOVIRUS INFECTIONS

水貂持续感染阿留申病细小病毒的研究 (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，免疫复合物疾病， 间质性和肾小球肾炎。主要靶细胞群 是巨噬细胞。感染是 增强抗病毒抗体，但在某种程度上限制在水平 单个细胞。这个项目的目的是阐明 病毒感染导致这种疾病的发病机制。 今年的研究摘要包括： * 对ADV非结构蛋白的抗体应答。 水貂ADV感染 针对病毒共有的氨基酸序列制造抗体 非结构蛋白NS 1、NS 2和NS 3。 虽然抗体的 也存在NS 1的独特区域，没有针对该独特区域的抗体。 检测到NS 2或NS 3区域。 * 允许感染中ADV蛋白的亚细胞定位。在 在感染后期的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，NS 1和NS 2的内含物。NS2（ca.（50%） 存在于细胞质中，表现出穿梭的一些特征 HIV rev蛋白。 * ADV限制性感染单核细胞系K562。 VP，NS 1和NS 2 抗体依赖性感染后，K562细胞核内出现 与CRFK的允许ADV感染不同， K562的细胞质ADV感染可使正常细胞周期阻滞， K562晚期，类似于CRFK中观察到的细胞周期破坏。 在ADV中未观察到IL-6蛋白的上调 感染K562细胞。 * 水貂外周血单个核细胞（PBMC）的ADV感染。 一 建立了水貂PBMC中ADV感染的研究体系。以下 从未感染貂纯化PBMC的ADV感染，几乎所有 单核细胞显示特异性胞质荧光。低百分比 在以后的时间点继续显示抗原的核定位， 表明病毒复制有限。 * 聚合酶链反应（PCR）与免疫反应的比较 电泳（CEP）在ADV感染诊断中的应用。 CEP被发现 比PCR更灵敏地检测商业中的ADV感染， 水貂牧场的环境 * 水貂ADV感染的细胞因子反应 测定细胞因子水平 用非放射性RNA酶对ADV感染水貂的淋巴组织进行了研究 保护试验（RPA）。未观察到IL-6的一致模式， 感染和未感染之间的IL-10、IL-1、IL-12 p35、IL-12 p40或TNF 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.