PATHOGENESIS OF ALEUTIAN DISEASE VIRUS INFECTION

水貂持续感染阿留申病细小病毒的研究 导致免疫调节紊乱，包括多克隆 高丙种球蛋白血症，浆细胞增多症，免疫复合物疾病， 间质性和肾小球肾炎以及极高水平的抗- ADV抗体。研究结果的范围类似于 细胞因子应答的Th 2模式和先前的工作暗示了 IL-6的可能作用 本项目的范围是阐明 ADV感染导致这种不寻常的疾病的机制。 先前用于抗体依赖性增强（ADE）的U937细胞 ADV感染的细胞系为2个单核细胞系的混合细胞，>90% K562和<10%U937。 纯细胞群实验表明， K562细胞通过ADE对ADV敏感，而U937细胞则不敏感。 感染 是通过Fc γ RII受体。K562细胞分泌IL-6， 对ADV感染的反应。 为了评估所选细胞因子在免疫系统中的作用， 与ADV感染相关的疾病，RT-PCR对来自 感染貂的组织IL-4、IL-5、IL-10、IL-10的简并引物 IL-12 p35和IL-12 p40基于来自人，小鼠， 猫、老鼠、牛和猪。获得适当大小的PCR产物， 被克隆了。 目前正在进行DNA测序。 为了鉴定病毒复制的早期靶细胞， 经鼻内（IN）和腹腔内（IP）感染ADV。 进行性疾病通过任一途径发展，但进展似乎 在感染后更慢。ADV DNA的最早证据是由 引流淋巴组织的PCR。 为了比较ADV蛋白的亚细胞定位， 允许性和限制性感染中的核酸，特异性抗血清 已经制备了非结构蛋白（NS）和衣壳蛋白， 与荧光染料结合。 不对称PCR已用于开发 用于荧光原位的荧光素化链特异性探针 杂交（FISH）。 同步细胞的初步工作与多- 通道荧光和共聚焦显微镜表明，NS 1和NS 2 共定位，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.