PATHOGENESIS OF ALEUTIAN DISEASE VIRUS INFECTION

阿留申水貂病细小病毒(ADV)持续感染水貂 引起免疫调节紊乱，包括多克隆 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和过高的抗-HBs水平 ADV抗体。这一系列发现类似于那些与 细胞因子反应的Th2模式和以前的工作表明 IL-6可能发挥的作用。本项目的范围是为了阐明 ADV感染导致这种不寻常的疾病的机制。 以往用于抗体依赖增强(ADE)的U937细胞 90%的ADV感染是由2个单核细胞株混合而成。 K562和10%U937。对纯细胞群体的实验表明 通过ADE，K562细胞对ADV敏感，而U937细胞不敏感。感染 是通过Fc-Gamma RII受体。IL-6是由K562细胞在 对ADV感染的反应。 为了评估选定的细胞因子在免疫中的作用 与ADV感染相关的疾病，对来自 受感染的水貂的组织。白介素4、白介素5、白介素10、白介素2的简并引物 12p35和IL-12p40基于人、小鼠、 猫、老鼠、牛和猪。获得了大小合适的聚合酶链式反应产物 已经被克隆了。DNA测序目前正在进行中。 为了确定病毒复制的早期目标细胞，水貂 鼻腔(IN)和腹膜(IP)感染ADV。 进展性疾病通过这两种途径发展，但进展似乎 在感染IN后速度变慢。ADV DNA的最早证据是由 对引流淋巴组织进行聚合酶链式反应。 为了比较ADV蛋白和ADV蛋白的亚细胞定位 允许性和限制性感染、特异性抗血清中的核酸 对于非结构(NS)和衣壳蛋白已经制备和 与荧光染料结合。不对称聚合酶链式反应已被用于开发 荧光原位荧光单链特异性探针 杂交(鱼)。多信元同步信元的初步研究 通道荧光和共聚焦显微镜显示NS1和NS2 共同定位，但NS和衣壳蛋白是空间分离的 在被允许感染的细胞的细胞核内。

Persistent infection of mink with Aleutian mink disease parvovirus (ADV) causes disturbances of immune regulation, including polyclonal hypergammaglobulinemia, plasmacytosis, immune complex disease, interstitial and glomerulonephritis and exceedingly high levels of anti- ADV antibodies. The spectrum of findings resembles those associated with a Th2 pattern of cytokine responses and previous work implicates a possible role for IL-6. The scope of this project is to elucidate mechanisms by which ADV infection results in this unusual disorder. The U937 cells used previously in antibody-dependent enhancement (ADE) of ADV infection were found to be mixture of 2 monocytic cell lines, >90% K562 and <10% U937. Experiments with pure cell populations indicated that K562, but not U937 cells were susceptible to ADV via ADE. Infection was via the Fc gamma RII receptor. IL-6 was elaborated by K562 cells in response to ADV infection. In order to evaluate the role of selected cytokines in the immune disorder associated with ADV infections, RT-PCR was done on mRNA from tissues from infected mink. Degenerate primers for IL-4, IL-5, IL-10, IL- 12 p35 and IL-12 p40 were based on consensus sequences from human, mouse, cat, rat, cow and pig. Appropriate sized PCR products were obtained and have been cloned. DNA sequencing is currently being done. In order to identify early target cells for viral replication, mink were infected both intranasally (IN) and intraperitoneally (IP) with ADV. Progressive disease develops by either route, but progression seems slower after IN infection. Earliest evidence of ADV DNA was localized by PCR to draining lymphoid tissues. In order to compare the subcellular localization of ADV proteins and nucleic acids in permissive and restricted infection, specific antisera for nonstructural (NS) and capsid proteins have been prepared and conjugated to fluorochromes. Asymmetric PCR has been used to develop fluoresceinated strand specific probes for fluorescence in situ hybridization (FISH). Preliminary work on synchronized cells with multi- channel epifluorescence and confocal microscopy suggests that NS1 and NS2 co-localize, but that NS and capsid proteins are spatially segregated within the nucleus of permissively infected cells.