PATHOGENESIS OF ALEUTIAN DISEASE VIRUS INFECTION

这个项目的范围是阐明涉及到的致病机制 水貂感染阿留申水貂病细小病毒(ADV) 在过去的一年里，我们开发了一种原位杂交的方法，使用 非放射性探测器。非放射性探测器提供了更清晰的图像 细胞形态比放射性探针强，尽管它们明显 不那么敏感。使用这两种探测类型，我们尝试识别更多 与ADV在成年水貂体内复制的靶细胞密切相关。我们以前的 研究表明，靶细胞在淋巴结中被发现，并具有 滤泡树突状细胞和巨噬细胞的分布。使用探测器 针对病毒粒子DNA隔离，我们将病毒粒子序列定位在 巨噬细胞和滤泡树突状细胞。但是，使用条件 特异的ADV mRNA、ADV复制可以最终定位为 巨噬细胞。 分离巨噬细胞和滤泡树突状细胞的尝试 被感染的水貂淋巴结产生的细胞数量少，活性差。 此外，ADV感染细胞的数量可以在 这些种群是通过原位杂交和类型 无法定义单元格。 对感染ADV的水貂的腹膜细胞也进行了研究。 含有AdV核酸的细胞很容易被鉴定。数量 含病毒粒子DNA的细胞约为含AdV mRNA的细胞的10倍。 其中一些细胞是吞噬细胞，支持巨噬细胞 是ADV的靶细胞。最后，贴壁的常驻腹膜细胞来自 正常水貂在体外感染ADV和其中的少量 通过产生ADV抗原来评估细胞复制ADV。 在其他工作中，我们开始研究ADV在体内的转录程序。 染病水貂肺组织中mRNA的Northern杂交分析 提示ADV在该组织中的转录与观察到的相似 在允许的体外感染中。从淋巴组织中提取的mRNA和 感染的成年水貂的腹膜渗出液细胞也有可检测到的ADV 信使核糖核酸，但数量较少。然而，所有的adv基因都出现了。 活在当下。

The scope of this project is to elucidate pathogenic mechanisms involved in infections of mink with Aleutian mink disease parvovirus (ADV). In the past year, we developed a method for in situ hybridization using nonradioactive probes. Nonradioactive probes provide a clearer picture of cellular morphology than radioactive probes, although they are noticeably less sensitive. Using both probe types, we have attempted to identify more closely the target cells for ADV replication in adult mink. Our previous work indicates the target cells are found in lymph nodes and have the distribution of follicular dendritic cells and macrophages. Using probes specific for virion DNA sequestration, we localized virion sequences in both macrophages and follicular dendritic cells. However, using conditions specific ADV mRNA, ADV replication could be conclusively localized only to macrophages. Attempts to isolate macrophages and follicular dendritic cells from infected mink lymph nodes yielded low numbers of cells with poor viability. Furthermore, extremely low numbers of ADV infected cells could be found in these populations ass assessed by in situ hybridization and the type of cell could not be defined. Cells from the peritoneal cavity of ADV infected mink were also studies. Cells containing ADV nucleic acid were readily identified. The number of cells containing virion DNA was about 10 times that of cells with ADV mRNA. Some of these cells were phagocytic, supporting the notion that macrophages are target cells for ADV. Finally, adherent resident peritoneal cells from normal mink were infected in vitro with ADV and a small number of these cells replicated ADV as assessed by the production of ADV antigens. In other work, we began to study the transcription program of ADV in vivo. Northern blot analysis of mRNA from the lungs of infected mink kits suggested that ADV transcription in this tissue is similar to that observed in permissive in vitro infections. mRNA from the lymph nodes and peritoneal exudate cells of infected adult mink also had detectable ADV mRNA, but the amount was low. Nevertheless, all ADV mRNA species appeared to be present.