PATHOGENESIS OF ALEUTIAN DISEASE VIRUS INFECTION

阿留申病病毒感染的发病机制

• 批准号: 5200391
• 负责人: M E BLOOM
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5200391
• 关键词: Parvoviridae; antibody receptor; capsid; chimeric proteins; glomerulonephritis; hyperglobulinemia; immune complex diseases; in situ hybridization; interstitial nephritis; laboratory mouse; microorganism immunology; open reading frames; polymerase chain reaction; protein biosynthesis; virus antigen; virus protein; virus replication

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.

阿留申梅克病细节病毒（ADV）持续感染貂皮 引起免疫调节的干扰，包括多克隆 高γ-球蛋白血症，浆细胞增多症，免疫复杂疾病， 间质和肾小球肾炎以及极高的抗抗炎 ADV抗体。发现的范围类似于与之相关的发现 细胞因子反应和先前工作的Th2模式暗示了 IL-6的可能作用。 该项目的范围是阐明 ADV感染导致这种异常疾病的机制。 以前用于抗体依赖性增强（ADE）的U937细胞 发现ADV感染是2种单核细胞系的混合物，> 90％ K562和<10％U937。 指示纯细胞群的实验 那个K562而不是U937细胞不容易通过ADE受到ADV的影响。 感染 通过FC Gamma RII受体。 IL-6由K562细胞详细说明 对ADV感染的反应。 为了评估选定细胞因子在免疫中的作用 与ADV感染相关的障碍，RT-PCR是在mRNA上进行的 来自感染貂皮的组织。 IL-4，IL-5，IL-10，IL-的退化引物 12个P35和IL-12 p40是基于人，小鼠的共识序列 猫，老鼠，牛和猪。获得适当尺寸的PCR产品，并 被克隆了。 目前正在进行DNA测序。 为了识别早期靶细胞以进行病毒复制，貂是 用adv感染了鼻内（IN）和腹膜内（IP）。 进行性疾病是通过两种途径发展的，但进展似乎 感染后慢。 ADV DNA的最早证据是由 PCR到排水淋巴组织。 为了比较ADV蛋白的亚细胞定位和 宽容和受限感染中的核酸，特异性抗血清 用于非结构（NS）和衣壳蛋白，已经准备好了，并且 与荧光染料共轭。 不对称PCR已用于发展 荧光特异性探针的原位荧光 杂交（鱼）。 与多种同步细胞的初步工作 通道的落荧光和共聚焦显微镜表明NS1和NS2 共定位，但是NS和衣壳蛋白在空间上被隔离 在允许感染细胞的核内。